Options Manual - Beckman Coulter

Options Manual - Beckman Coulter
COULTER® EPICS® ALTRA™ Flow Cytometer
COULTER EPICS ALTRA HyPerSort™ System
Flow Cytometer
PN 4237470AA (August 2010)
Beckman Coulter, Inc.
250 S. Kraemer Blvd.
Brea, CA 92821
Options Manual
WARNINGS AND PRECAUTIONS
READ ALL PRODUCT MANUALS AND CONSULT WITH BECKMAN COULTER-TRAINED PERSONNEL BEFORE ATTEMPTING
TO OPERATE INSTRUMENT. DO NOT ATTEMPT TO PERFORM ANY PROCEDURE BEFORE CAREFULLY READING ALL
INSTRUCTIONS. ALWAYS FOLLOW PRODUCT LABELING AND MANUFACTURER’S RECOMMENDATIONS. IF IN DOUBT AS
TO HOW TO PROCEED IN ANY SITUATION, CONTACT YOUR BECKMAN COULTER REPRESENTATIVE.
HAZARDS AND OPERATIONAL PRECAUTIONS AND LIMITATIONS
WARNINGS, CAUTIONS, and IMPORTANTS alert you as follows:
WARNING - Can cause injury.
CAUTION - Can cause damage to the instrument.
IMPORTANT - Can cause misleading results.
BECKMAN COULTER, INC. URGES ITS CUSTOMERS TO COMPLY WITH ALL NATIONAL HEALTH AND SAFETY
STANDARDS SUCH AS THE USE OF BARRIER PROTECTION. THIS MAY INCLUDE, BUT IT IS NOT LIMITED TO,
PROTECTIVE EYEWEAR, GLOVES, AND SUITABLE LABORATORY ATTIRE WHEN OPERATING OR MAINTAINING THIS OR
ANY OTHER AUTOMATED LABORATORY ANALYZER.
WARNING Risk of operator injury if:
r All doors, covers and panels are not closed and secured in place prior to and during instrument operation.
r The integrity of safety interlocks and sensors is compromised.
r Instrument alarms and error messages are not acknowledged and acted upon.
r You contact moving parts.
r You mishandle broken parts.
r Doors, covers and panels are not opened, closed, removed and/or replaced with care.
r Improper tools are used for troubleshooting.
To avoid injury:
r Keep doors, covers and panels closed and secured in place while the instrument is in use.
r Take full advantage of the safety features of the instrument. Do not defeat safety interlocks and sensors.
r Acknowledge and act upon instrument alarms and error messages.
r Keep away from moving parts.
r Report any broken parts to your Beckman Coulter Representative.
r Open/remove and close/replace doors, covers and panels with care.
r Use the proper tools when troubleshooting.
CAUTION System integrity might be compromised and operational failures might occur if:
r This equipment is used in a manner other than specified. Operate the instrument as instructed in the Product Manuals.
r You introduce software that is not authorized by Beckman Coulter into your computer. Only operate your system’s
computer with software authorized by Beckman Coulter.
r You install software that is not an original copyrighted version. Only use software that is an original copyrighted
version to prevent virus contamination.
IMPORTANT If you purchased this product from anyone other than Beckman Coulter or an authorized Beckman Coulter
distributor, and, if it is not presently under a Beckman Coulter service maintenance agreement, Beckman Coulter cannot
guarantee that the product is fitted with the most current mandatory engineering revisions or that you will receive the most
current information bulletins concerning the product. If you purchased this product from a third party and would like
further information concerning this topic, call your Beckman Coulter Representative.
REVISION STATUS
Initial Release 2/00
Software Version 1.0
Issue AA, 8/10
Software Version 1.0.
Updates were made to the company corporate address.
Note: Changes that are part of the most recent revision are indicated in text by a bar in the
margin of the amended page.
This document applies to the latest software listed and higher versions. When a subsequent software version
changes the information in this document, a new issue will be released to the Beckman Coulter website. For
labeling updates, go to www.beckmancoulter.com and download the most recent manual or system help for
your instrument.
PN 4237470AA
iii
REVISION STATUS
iv
PN 4237470AA
CONTENTS
LEGAL NOTICES
REVISION STATUS, iii
CONTENTS, v
INTRODUCTION, xi
HOW TO USE YOUR COULTER EPICS ALTRA FLOW CYTOMETER MANUALS, xi
ABOUT THE OPTIONS MANUAL, xi
HyPerSort™ System (HPSS) Option, xii
CONVENTIONS, xii
1
2
PN 4237470AA
SAFETY PRECAUTIONS, 1-1
1.1
BIOLOGICAL HAZARDS, 1-1
1.2
LASER SAFETY PRECAUTIONS, 1-1
Radiation Hazards, 1-2
AIR-COOLED LASERS, 2-1
2.1
GENERAL INFORMATION, 2-1
Installation Requirements, 2-1
Floor Loading and Space, 2-1
Heat and Ventilation, 2-1
Operation, 2-1
Aligning the Mirrors, 2-1
Mirror Alignment Procedure, 2-2
Cleaning and Other Maintenance, 2-16
2.2
AIR-COOLED ARGON LASERS, 2-17
Electrical Power, 2-17
Startup, 2-17
Shutdown, 2-18
2.3
RED HeNe LASERS, 2-19
Electrical Power, 2-19
Controls and Indicators, 2-19
Laser Head, 2-19
Startup, 2-20
Shutdown, 2-21
2.4
GREEN HeNe LASERS, 2-22
Electrical Power, 2-22
Controls and Indicators, 2-22
Power Supply, 2-22
Startup, 2-24
Shutdown, 2-25
-v
CONTENTS
2.5
3
-vi
HeCd LASERS, 2-26
Electrical Power, 2-26
Controls and Indicators, 2-26
Power Supply, 2-26
Laser Head, 2-28
Startup, 2-29
Procedure, 2-29
Shutdown, 2-30
WATER-COOLED LASERS, 3-1
3.1
GENERAL INFORMATION, 3-1
Installation Requirements, 3-1
Heat and Ventilation, 3-1
Operation, 3-1
Changing Laser Wavelength, 3-1
Aligning the Mirrors, 3-2
Mirror Alignment Procedure, 3-2
Cleaning and Other Maintenance, 3-13
3.2
ENTERPRISE II SERIES LASERS, 3-14
Floor Loading and Space, 3-14
Electrical Power, 3-14
Cooling Water, 3-15
Controls and Indicators, 3-15
Power Supply, 3-15
Remote Control Module, 3-16
Laser Head, 3-17
Startup, 3-18
Set Laser Power, 3-19
Shutdown, 3-19
3.3
300 SERIES LASERS, 3-20
Floor Loading and Space, 3-20
Electrical Power, 3-21
Cooling Water, 3-21
Controls and Indicators, 3-21
Power Supply, 3-21
Remote Control Module, 3-22
Laser Head, 3-23
Startup, 3-24
Set Aperture, 3-27
Set Laser Power, 3-27
Tune Laser for Maximum Power, 3-27
Shutdown, 3-28
PN 4237470AA
CONTENTS
3.4
4
5
6
STORAGE OPTIONS, 4-1
4.1
GENERAL INFORMATION, 4-1
Magnetic Storage Media Care, 4-1
4.2
Zip™ DRIVE, 4-1
4.3
RECORDABLE CD-ROM DRIVE, 4-1
Easy CD Creator, 4-2
Easy CD-Creator Installation Procedures, 4-2
4.4
jaz® DRIVE, 4-2
Autoclone™ SORTING OPTION, 5-1
5.1
INTRODUCTION, 5-1
5.2
INSTALLATION REQUIREMENTS, 5-1
Installation Calibration Procedure, 5-1
5.3
STARTUP, 5-4
5.4
SORT MATRIX, 5-8
5.5
SORTING, 5-15
5.6
CALIBRATION, 5-21
Fine Tune Calibration, 5-22
Adjust Autoclone Sorting Option Speed, 5-26
Noisy Movement, 5-26
Missing Wells, 5-27
GATED AMP OPTION, 6-1
6.1
PN 4237470AA
SPECTRUM LASER, 3-28
Floor Loading and Space, 3-29
Electrical Power, 3-29
Cooling Water, 3-29
Controls and Indicators, 3-29
Power Supply, 3-29
Remote Control Module, 3-31
Laser Head, 3-32
Startup, 3-33
Set Laser Power, 3-34
Tune Laser for Maximum Power, 3-35
Shutdown, 3-35
WHAT THIS CHAPTER EXPLAINS, 6-1
-vii
CONTENTS
7
8
6.2
FEATURES OF THE GATED AMP, 6-1
Time-Domain Gating, 6-1
Lower Beam Lockout, 6-1
Dual Laser Applications, 6-1
Example: DAPI/FITC for Viable DNA and Immunofluorescence, 6-1
Example: Calcium Flux with Indo-1 and Two-Color Cell-Surface Markers, 6-1
Example: Hoechst/Chromomycin for Dual-Color Chromosome Sorting, 6-2
Example: Simultaneous Four-Color Clone Studies, 6-2
6.3
IMAGING OPTICS ALIGNMENT, 6-2
6.4
SET UP GATED AMP SCREENS, 6-6
Gated Amp Assignment Screen, 6-6
Gated Amp Control Screen, 6-8
Gated Amp Control Screen Settings, 6-10
Fluorescence Compensation Combinations, 6-10
Software Selectable Amplifier Speed/Sensitivity, 6-11
6.5
CHECK ALIGNMENT, 6-11
6.6
ADJUST WINDOW WIDTH, 6-13
LARGE PARTICLE SORTING OPTION, 7-1
7.1
DESCRIPTION, 7-1
7.2
COMPONENTS, 7-1
7.3
INSTALL THE LARGE PARTICLE SORTING OPTION, 7-1
Procedure, 7-1
7.4
REPLACE THE SAMPLE TUBING ASSEMBLY, 7-12
Procedure, 7-12
7.5
RUN SAMPLES, 7-17
Install Large Particle Sorting option, 7-17
Select the Flow Cell Tip, 7-17
Select Settings, 7-17
Run Sample, 7-18
FORWARD FLUORESCENCE DETECTOR OPTION, 8-1
8.1
DESCRIPTION, 8-1
8.2
INSTALL THE FORWARD FLUORESCENCE DETECTOR OPTION, 8-1
INDEX, INDEX-1
TRADEMARKS
-viii
PN 4237470AA
CONTENTS
ILLUSTRATIONS
1.1
1.2
1.3
1.4
1.5
1.6
1.7
1.8
1.9
1.10
1.11
1.12
2.1
2.2
2.3
2.4
2.5
2.6
3.1
3.2
3.3
3.4
3.5
3.6
3.7
3.8
3.9
3.10
6.1
6.2
PN 4237470AA
Argon Laser, Manufacturer’s Warning Labels, 1-3
Red Helium-Neon (HeNe) Laser, Manufacturer’s Warning Labels, 1-4
Green HeNe Laser, Manufacturer’s Warning Label, 1-4
Helium Cadmium (HeCd) Laser, Manufacturer’s Warning Labels, 1-5
Coherent® Innova® Enterprise II Series Laser, Manufacturer’s’ Warning
Labels, 1-5
Coherent Innova 300 Series Laser, Manufacturer’s Warning Labels, 1-6
Coherent Innova 70 Spectrum Laser, Manufacturer’s Warning Labels, 1-6
Cytometer Sensing Area, Laser Warning Labels, 1-7
Forward Fluorescence Detector Option Sensing Area, Laser Warning Labels, 1-8
Cytometer Sensing Compartment’s Cover, Laser Warning Labels, 1-9
Cytometer Outer Cover, Laser Classification Label, 1-9
Cytometer Rear and Left Cover Laser Warning Labels and Laser Certification
Label, 1-10
Laser Arrangement - Air-Cooled Laser, Example, 2-2
Laser Head - Red HeNe Laser, 2-19
Power Supply - Green HeNe Laser When Used As a Third Laser, 2-23
Laser Head - Green HeNe Laser, 2-23
Power Supply - HeCd Lasers, 2-27
Laser Head - HeCd Lasers, 2-28
Laser Arrangement - Water-Cooled Lasers in a Three Laser Configuration, 3-2
Power Supply - Enterprise II Series Lasers, 3-16
Remote Control Module - Enterprise II Series Lasers, 3-16
Laser Head - Enterprise II Series, 3-17
Power Supply - 300 Series Lasers, 3-22
Remote Control Module - 300 Series Lasers, 3-22
Laser Head - 300 Series Lasers, 3-24
Power Supply - Spectrum Laser, 3-30
Remote Control Module - Spectrum Laser, 3-31
Laser Head - Spectrum Laser, 3-32
Gated Amp Assignment Screen, 6-6
Gated Amp Control Screen, 6-8
-ix
CONTENTS
TABLES
2.1
2.2
3.1
3.2
3.3
3.4
3.5
3.6
3.7
7.1
-x
Power Supply Control Settings - HeCd Lasers, 2-27
Power Supply Indicators - HeCd Lasers, 2-28
Cooling Water Specifications - Enterprise II Series Lasers, 3-15
Power Supply Controls/Indicators - Enterprise II Series Lasers, 3-15
Remote Control Module - Enterprise II Series Lasers, 3-17
Cooling Water Specifications - 300 Series Lasers, 3-21
Remote Control Module - 300 Series Lasers, 3-23
Cooling Water Specifications - Spectrum Laser, 3-29
Power Supply and Remote Control Module Controls and Indicators - Spectrum Laser, 3-30
Typical Settings for Large Particle Sorting, 7-17
PN 4237470AA
INTRODUCTION
This introductory section contains the following topics:
r
How to use your COULTER® EPICS® ALTRA™ and ALTRA HyPerSort™ System flow
cytometer manuals
r
About the Options manual
r
Conventions.
HOW TO USE YOUR COULTER EPICS ALTRA FLOW CYTOMETER MANUALS
Use the Reference manual for in-depth information about:
r
What the instrument does
r
The methods it uses
r
Its specifications
r
Information about installation.
r
Safety.
Use the Operator’s Guide for the day-to-day running of your instrument:
Read the Controls and Indicators chapter to become familiar with the different parts of your
system. Read the Software chapter to become familiar with how to work with the software.
Then go through the detailed step-by-step procedures of daily startup, running controls and
samples, reviewing data, sorting and shutdown. Appendices include information on defining
protocols, managing data, setting up prism, ratio and time parameters, working with
metafiles, setting display colors, using panelyzer mode, and using the time of flight and pulse
pile up features.
Use the Special Procedures and Troubleshooting manual for weekly cleaning and
replacement/adjustment procedures and troubleshooting information.
Use the Options manual for information on how to work with any of the options on your
instrument.
Use the FlowCentre™ Multimedia Workstation Getting Started manual for information how
to interface with the Microsoft® Windows™ 98 software.
See the Documentation page on the back cover of this manual for the contents of each
manual. It can help you to determine quickly in what manual is the information you need.
ABOUT THE OPTIONS MANUAL
This manual includes instructions for the basic operation of the options available with the
ALTRA flow cytometer. Additional manuals may be supplied with the option by the original
manufacturer of the option. Refer to the manufacturer’s manual for complete operating
instructions and the use of advanced features.
Some procedures in the ALTRA flow cytometer manuals may change when an option is
installed. See the chapter in this manual covering the option for these changes.
Some options such as Printers are not contained in this manual. Refer to the manufacturer’s
manual for operating instructions.
PN 4237470AA
-xi
INTRODUCTION
CONVENTIONS
s
Chapter 1, Safety Precautions
Contains biological and laser safety information.
s
Chapter 2, Air-Cooled Lasers
Contains descriptions and basic operating instructions for the available air-cooled lasers:
Argon, red HeNe, green HeNe, and HeCd lasers.
s
Chapter 3, Water-Cooled Lasers
Contains descriptions and basic operating instructions for the available water-cooled
lasers: Coherent® Innova® Enterprise II, 300 Series and Spectrum lasers.
s
Chapter 4, Storage Options
Contains descriptions of the different storage options available and any procedures
needed such as installation or archiving.
s
Chapter 5, Autoclone™ Sorting Option
Describes how to perform sorting with the Autoclone sorting option.
s
Chapter 6, Gated Amp Option
Describes the features of the Gated Amp option.
s
Chapter 7, Large Particle Sorting Option
Describes the features of the Large Particle Sorting option.
s
Chapter 8, Forward Fluorescence Detector Option
Describes the features of the Forward Fluorescence Detector option.
s
Index
Use the Index to easily locate specific information in this manual.
HyPerSort™ System (HPSS) Option
Information for the COULTER EPICS ALTRA HyPerSort System flow cytometer is included
in the ALTRA flow cytometer Reference, Operator’s Guide, and Special Procedures and
Troubleshooting manuals.
CONVENTIONS
This manual uses the following conventions:
Italics indicate screen messages.
Bold indicates a menu item.
Courier font indicates text you have to type using the keyboard.
ì indicates a key (such as Û).
-xii
PN 4237470AA
INTRODUCTION
CONVENTIONS
ì+ì indicates that the two keys listed (such as Þ+Ê) are linked for a specific
function and must be pressed in this sequence:
1. Press down on the first key listed and while continuing to press it, press down on the
second key listed.
2. Release both keys at the same time.
ì ì indicates to press and release the first key listed then press and release the next key
listed. For example, y Û.
Screen tt Analysis
Select the Screen item on the Analysis menu.
[OKAY]
Use the mouse to click on the screen button labeled [OKAY].
É through Ô
PN 4237470AA
Special function keys.
-xiii
INTRODUCTION
CONVENTIONS
-xiv
PN 4237470AA
1SAFETY PRECAUTIONS 1
1.1
BIOLOGICAL HAZARDS
The area around the system may contain materials that create a biological hazard to
laboratory personnel. Process samples with the appropriate precautions to prevent exposure
to biohazardous material. The extent of biohazard containment depends on the degree of risk
involved, according to the type and classification of the sample.
Provide for decontamination of the instrument and associated laboratory equipment, supplies
and waste. Decontaminate immediately after examining potentially biohazardous samples.
For help in establishing procedures and facilities for biohazard control, refer to the guidelines
presented in the following publications. Always obtain the latest edition of these publications.
1.2
r
Biohazards Safety Guide National Institutes of Health, 1974.
r
Centers for Disease Control. Acquired Immune Deficiency Syndrome (AIDS):
Precautions for clinical laboratory staffs. Morbidity and Mortality Weekly Report
31(43):577-579, 1982.
r
Classification of Etiologic Agents on the Basis of Hazard, 3d ed., Centers for Disease
Control, U.S. Public Health Service, June 1972.
r
National Committee for Clinical Laboratory Standards. Protection of laboratory workers
from infectious disease transmitted by blood, body fluids and tissue; Tentative guideline.
NCCLS Document M29-T. Villanova, PA.: NCCLS; 1989.
r
National Committee for Clinical Laboratory Standards. Clinical applications of flow
cytometry: quality assurance of peripheral blood lymphocytes; Tentative Guideline.
NCCLS Document H42-T. Villanova, PA.: NCCLS; pp. 12, 13, 37, 38. 1992.
r
MMWR, Vol. 46, No. RR-2, 1997. Revised guidelines for performing CD4+ T-cell
determinations in persons with Human Immunodeficiency Virus (HIV): Laboratory
Safety Centers for Disease Control; pp. 3-4. Jan 10, 1997.
LASER SAFETY PRECAUTIONS
The laser is a unique light source that shows characteristics different from conventional light
sources. The safe use of the laser depends upon familiarity with the instrument and with the
properties of coherent, intense beams of light.
The beam can cause eye damage or blindness and instrument damage. There is enough power
from the laser to ignite substances placed in the beam path, even at some distance. The beam
might also cause damage if contacted indirectly from reflective surfaces (specular reflection).
For these reasons follow the precautions that follow.
WARNING Use of controls or adjustments or performance of procedures other than those specified herein
may result in hazardous radiation exposure.
PN 4237470AA
1.
Carefully read the safety instructions in the manual for the laser.
2.
Do not use the lasers in the presence of flammables or explosives; these include volatile
substances such as alcohol, solvents, and ether.
3.
Limit access to the instrument. Keep the instrument out of the hands of inexperienced
and untrained personnel.
1-1
SAFETY PRECAUTIONS
LASER SAFETY PRECAUTIONS
4.
Never look directly into the laser light source or at scattered laser light from any
reflective surface. Never look down the beam into the source.
5.
As a precaution against accidental exposure to the output beam or to its reflection, those
performing service or maintenance procedures on the system should wear safety glasses
as required by the wavelength being used.
6.
The intensity of the beam can cause flesh burns. Avoid direct exposure and specular reflections.
7.
Post warning signs in the area of the laser beam to alert those present.
8.
Advise all those using the laser of these precautions.
Radiation Hazards
In its design and manufacture of the ALTRA flow cytometer, Beckman Coulter Inc. has
complied with the requirements governing the use and application of a laser as stipulated in
regulatory documents issued by the:
r
U.S. Department of Health and Human Services
r
Center for Devices and Radiological Health (CDRH).
In compliance with these regulatory documents, every measure has been taken to ensure the
health and safety of users and laboratory personnel from the possible dangers of laser use.
WARNING Risk of personal injury. This instrument contains components dangerous to the operator. If any
attempt has been made to defeat a safety feature, or if this instrument does not perform as listed in this
manual, disconnect power and call your Beckman Coulter Representative.
CDRH-approved warning labels are placed near or on those covers that when removed might
expose laser radiation. Only remove those covers when performing service and maintenance
on the system as instructed in the COULTER EPICS ALTRA Flow Cytometer Special
Procedures and Troubleshooting manual.
See Figures 1.1 through 1.12 for the labels and their locations on the lasers:
1-2
r
Figure 1.1, Argon Laser, Manufacturer’s Warning Labels
r
Figure 1.2, Red Helium-Neon (HeNe) Laser, Manufacturer’s Warning Labels.
r
Figure 1.3, Green HeNe Laser, Manufacturer’s Warning Label.
r
Figure 1.4, Helium Cadmium (HeCd) Laser, Manufacturer’s Warning Labels
r
Figure 1.5, Coherent® Innova® Enterprise II Series Laser, Manufacturer’s’ Warning
Labels
r
Figure 1.6, Coherent Innova 300 Series Laser, Manufacturer’s Warning Labels
r
Figure 1.7, Coherent Innova 70 Spectrum Laser, Manufacturer’s Warning Labels
r
Figure 1.8, Cytometer Sensing Area, Laser Warning Labels.
r
Figure 1.9, Forward Fluorescence Detector Option Sensing Area, Laser Warning Labels
r
Figure 1.10, Cytometer Sensing Compartment’s Cover, Laser Warning Labels
r
Figure 1.11, Cytometer Outer Cover, Laser Classification Label.
r
Figure 1.12, Cytometer Rear and Left Cover Laser Warning Labels and Laser
Certification Label
PN 4237470AA
SAFETY PRECAUTIONS
LASER SAFETY PRECAUTIONS
When performing service or maintenance on the system with the covers open and the
interlock defeated, observe the following:
WARNING Reflections of the laser beam from a shiny object, such as a screwdriver, or direct viewing of the
laser beam can severely damage your eyes. Pay attention to the warning labels in the sampling
compartment. Do not wear any jewelry that might reflect the laser beam. Do not observe the intersection of
the laser beam and the flow chamber directly. To observe this point, use the video camera and graphics
screen at the cytometer. Wear laser safety glasses that protect against the wavelengths in use.
Replace the covers before resuming normal operation.
Figure 1.1 Argon Laser, Manufacturer’s Warning Labels
LASER RADIATION WHEN OPEN
AVOID DIRECT EXPOSURE
TO BEAM
Cyonics
a division of
uniphase
163 BAYPOINTE PKWY
SAN JOSE, CA 95134
MODEL
SERIAL NO.
MANUFACTURED
THIS LASER PRODUCT COMPLIES
WITH 21 CFR 1040 AS APPLICABLE
LASER RADIATION-AVOID
DIRECT EXPOSURE TO BEAM
ARGON : .50 WATT CW.
CLASS IIIb LASER PRODUCT
LASER RADIATION
IS EMITTED FROM
THIS APERTURE
AVOID EXPOSURE
7470028A
PN 4237470AA
1-3
1
SAFETY PRECAUTIONS
LASER SAFETY PRECAUTIONS
Figure 1.2 Red Helium-Neon (HeNe) Laser, Manufacturer’s Warning Labels
ME
LASER APERTURE
CAUTION-LASER LIGHT WHEN OPEN
OPEN
SHUTTER
CLOSE
E
LL
05
S
IO
GR
T
91
HP
-9
CE
-L
G
MF
S/N
CA
SB
RL
AD
CA
R
SE
LA T
IS UC ITH
TH ROD S W
P LIE DRH
N
MP S/C
CO HH IATIO NCE
D D MA S
RA OR RD
RF DA
J
PE TAN CFR R 93
S 21
PTE :19
HA -1
B-C 825
SU IEC
D
AN
NIA
OR
LIF
8
00
92
M
HT
LIG BEA DRH) 93)
TO (C
ER
19
LAS SURE UCT 825-1:
OD
C
PO
EX SER PR T (IE 2.8 nm
UC 63
OID LA
AV S IIIb PROD AT
R
UM
CL
CL
SE
AS
IM
LA
AX
S 3B mW M
30
AS
Laser
shutter
MELLES GRIOT
05 - LHP - 991
MFG
LASER LIGHT
AVOID EXPOSURE TO BEAM
S/N
CLASS IIIb LASER PRODUCT (CDRH)
CLASS 3B LASER PRODUCT (IEC 825-1:1993)
30mW MAXIMUM AT 632.8 nm
7469005C
THIS LASER
PRODUCT
COMPLIES WITH
DHHS/CDRH
RADIATION
PERFORMANCE
STANDARDS
21 CFR
SUB-CHAPTER J
AND IEC 825-1 :1993
CE
CARLSBAD CALIFORNIA 92008
Figure 1.3 Green HeNe Laser, Manufacturer’s Warning Label
AVOID EXPOSURE - Laserlight
is emitted from this aperture.
OPEN
SHUTTER
CLOSE
93
GR
05
-L
-1
1
38
R
SE
LA CT H
IS
U
IT
TH ROD S W H
P LIE DR
MP S/C ON E
CO HH IATI NC
D D MA S
RA R RD
RFO DA
J
PE TAN CFD R
3
S 21
PTE :199
HA 5-1
B-C C 82
U
S IE
D
AN
CE
G
MF
S/N
Laser
shutter
M
HT
LIG BEA DRH) 93)
TO (C
ER
19
LAS SURE UCT 825-1:
PO PROD (IEC nm
EX
R
T
3.5
OID LASE UC
AV S IIIb PROD AT 54
CL
CL
R
UM
AS
SE
IM
LA
AX
S 3B W M
5m
AS
MELLES GRIOT
05 - LGR - 193 - 381
MFG
LASER LIGHT
AVOID EXPOSURE TO BEAM
S/N
CLASS IIIb LASER PRODUCT (CDRH)
CLASS 3B LASER PRODUCT (IEC 825-1:1993)
5mW MAXIMUM AT 543.5 nm
CARLSBAD CALIFORNIA 92008
THIS LASER
PRODUCT
COMPLIES WITH
DHHS/CDRH
RADIATION
PERFORMANCE
STANDARDS
21 CFR
SUB-CHAPTER J
AND IEC 825-1 :1993
CE
7469006C
1-4
PN 4237470AA
SAFETY PRECAUTIONS
LASER SAFETY PRECAUTIONS
Figure 1.4 Helium Cadmium (HeCd) Laser, Manufacturer’s Warning Labels
INVISIBLE LASER RADIATION-AVOID
DIRECT EXPOSURE TO BEAM
LESS THAN
50.0 MW
HeCd
CLASS lllb
LASER PRODUCT
DANGER
LASER LIGHT WHEN OPEN
AVOID DIRECT EXPOSURE TO BEAM
INVISIBLE RADIATION
IS EMITTED FROM
THIS APERTURE
AVOID EXPOSURE
5965033A
Figure 1.5 Coherent® Innova® Enterprise II Series Laser, Manufacturer’s’ Warning Labels
DANGEROUS VOLTAGE
UNDER THIS COVER
VISIBLE AND
INVISIBLE LASER RADIATION
IS EMITTED FROM THIS APERTURE
AVOID EXPOSURE
VISIBLE AND INVISIBLE LASER RADIATION
AVOID EYE OR SKIN EXPOSURE TO
DIRECT OR SCATTERED RADIATION.
ARGON ION LASER
2 WATTS MAX.* CW
7003159A
PN 4237470AA
*PRACTICAL LIMIT
CLASS IV LASER PRODUCT
1-5
1
SAFETY PRECAUTIONS
LASER SAFETY PRECAUTIONS
Figure 1.6 Coherent Innova 300 Series Laser, Manufacturer’s Warning Labels
VISIBLE AND INVISIBLE LASER RADIATION
AVOID EYE OR SKIN EXPOSURE TO
DIRECT OR SCATTERED RADIATION.
ARGON ION LASER
12 WATTS MAX.* CW
*PRACTICAL LIMIT
VISIBLE AND
INVISIBLE LASER RADIATION
IS EMITTED
FROM THIS APERTURE
CLASS IV LASER PRODUCT
AVOID EXPOSURE
7003160A
Figure 1.7 Coherent Innova 70 Spectrum Laser, Manufacturer’s Warning Labels
VISIBLE AND
INVISIBLE LASER RADIATION
IS EMITTED
FROM THIS APERTURE
AVOID EXPOSURE
VISIBLE LASER LIGHTAVOID EXPOSURE TO
DIRECTOR SCATTERED LIGHT
7003161A
1-6
ARGON AND KRYPTON
MIXED GAS LASER
8 WATTS MAX. CW/MULTILINE OUTPUT.
PRACTICAL LIMT
CLASS IV LASER PRODUCT
PN 4237470AA
SAFETY PRECAUTIONS
LASER SAFETY PRECAUTIONS
Figure 1.8 Cytometer Sensing Area, Laser Warning Labels
VISIBLE AND/OR INVISIBLE
LASER RADIATION WHEN OPEN
AND INTERLOCK DEFEATED
AVOID EYE OR SKIN
EXPOSURE TO DIRECT OR
SCATTERED RADIATION
NG
ER
N AM
TIO BE T
DIA TO DUC
RA E
R SU R P R O
SE
R
L A XPO A SE
E
L
OID 3B
AV ASS
CL
Interlock
defeat bar
DA
DA
NG
ER
LASER RADIATION
AVOID EXPOSURE TO BEAM
CLASS 3B LASER PRODUCT
CL
AS
S
1
LA
SE
R
PR
OD
UC
T
AVOID EXPOSURE
VISIBLE AND/OR INVISIBLE
LASER RADIATION IS EMITTED
FROM THIS APERTURE
CLASS 1 LASER PRODUCT
7469015C
PN 4237470AA
1-7
1
SAFETY PRECAUTIONS
LASER SAFETY PRECAUTIONS
Figure 1.9 Forward Fluorescence Detector Option Sensing Area, Laser Warning Labels
VISIBLE AND/OR INVISIBLE
LASER RADIATION WHEN OPEN
AND INTERLOCK DEFEATED
N
DA
AVOID EYE OR SKIN
EXPOSURE TO DIRECT OR
SCATTERED RADIATION
R
DA
N
G
ER
N AM
TIO BE CT
DIA TO
U
RA RE ROD
R
P
SU
SE
LA XPO SER
E
LA
OID 3B
AV SS
CLA
Interlock
defeat bar
GE
LASER RADIATION
AVOID EXPOSURE TO BEAM
CLASS 3B LASER PRODUCT
AVOID EXPOSURE
VISIBLE AND/OR INVISIBLE
LASER RADIATION IS EMITTED
FROM THIS APERTURE
CLASS 1 LASER PRODUCT
7469024C
1-8
PN 4237470AA
SAFETY PRECAUTIONS
LASER SAFETY PRECAUTIONS
Figure 1.10 Cytometer Sensing Compartment’s Cover, Laser Warning Labels
VISIBLE AND/OR INVISIBLE
LASER RADIATION WHEN OPEN
AVOID EYE OR SKIN
EXPOSURE TO DIRECT OR
SCATTERED RADIATION
LASER RADIATION
AVOID EYE OR SKIN EXPOSURE TO
DIRECT OR SCATTERED RADIATION
CLASS 4 LASER PRODUCT
Latch
LASER RADIATION
AVOID EYE OR SKIN EXPOSURE TO
DIRECT OR SCATTERED RADIATION
CLASS 4 LASER PRODUCT
VISIBLE AND/OR INVISIBLE
LASER RADIATION WHEN OPEN
AVOID EYE OR SKIN
EXPOSURE TO DIRECT OR
SCATTERED RADIATION
7469004C
Figure 1.11 Cytometer Outer Cover, Laser Classification Label
7469012A
CLASS 1 LASER PRODUCT
PN 4237470AA
1-9
1
SAFETY PRECAUTIONS
LASER SAFETY PRECAUTIONS
Figure 1.12 Cytometer Rear and Left Cover Laser Warning Labels and Laser Certification Label
VISIBLE AND/OR INVISIBLE
LASER RADIATION WHEN OPEN
LASER RADIATION
AVOID EYE OR SKIN EXPOSURE TO
DIRECT OR SCATTERED RADIATION
CLASS 4 LASER PRODUCT
AVOID EYE OR SKIN
EXPOSURE TO DIRECT OR
SCATTERED RADIATION
THIS LASER PRODUCT CONFORMS
TO THE PROVISIONS OF 21 CFR.
SUBCHAPTER J, SECTIONS
1040.10 AND 1040.11.
Manufactured:
DATE
Beckman Coulter, Inc.
250 S. Kraemer Blvd.
Brea, CA 92821
by:
7469003C
1-10
PN 4237470AA
2AIR-COOLED LASERS 2
2.1
GENERAL INFORMATION
The following air-cooled lasers are available as options on the ALTRA flow cytometer:
r
Cyonics Air-cooled Argon laser (hereafter called the air-cooled Argon laser)
r
Melles Griot or Uniphase Red Helium Neon (HeNe) laser (hereafter called the Red HeNe
laser)
r
Melles Griot Green Helium Neon (HeNe) laser (hereafter called the Green HeNe laser)
r
Omnichrome Helium-Cadium (HeCd) laser (hereafter called the HeCd laser).
The information under this section (Heading 2.1, GENERAL INFORMATION) is generic and
applies to all the air-cooled lasers listed above. Information specific to each laser listed above
is included in a separate section in this chapter.
Installation Requirements
Do not unpack or install the option yourself. The option must be unpacked and installed by a
Beckman Coulter Representative. Certain internal adjustments of the lasers and the
instrument may be required at the time of installation.
Refer to the Installation chapter in the ALTRA flow cytometer Reference manual for other
installation requirements.
Floor Loading and Space
The dimensions and weight are the same as the standard ALTRA flow cytometer. Refer to the
ALTRA flow cytometer Reference manual.
Heat and Ventilation
The ALTRA flow cytometer dissipates approximately 2,400 W (8,191 Btu/hour) of heat.
Depending upon the type and power of the associated lasers, the total heat dissipation can
increase considerably. Be sure to consider this to ensure that the room where the ALTRA flow
cytometer is located is properly ventilated and can remove the generated heat.
For example: a typical air-cooled 15 mW argon laser can dissipate an additional 2,000 W
(6,824 Btu/hour) of heat. An air-cooled Omnichrome Model 74 HeCd laser dissipates about
100 W (341 Btu/hour) of heat. See the laser manufacturer’s specifications for details.
The system must be operated in an air-conditioned space. Keep room temperature between
18°C and 29°C (64°F and 85°F).
Operation
This chapter contains the minimum information necessary for operation with the ALTRA
flow cytometer.
For complete operating instructions, refer to the specific laser manufacturer’s manual.
Aligning the Mirrors
To align the mirrors you need to be aware of the arrangement of the lasers, the path the laser
beam follows, and the filter configuration. One example is shown in the mirror alignment
procedure that follows.
PN 4237470AA
2-1
AIR-COOLED LASERS
GENERAL INFORMATION
With the addition of lasers, the orientation of the lasers are changed. All lasers point toward
the right side of the instrument into an optical rail supporting three dichroic alignment
mirrors, one for each laser (see Figure 2.1). The dichroic alignment mirrors reflect the laser
light into the sensing area.
Figure 2.1 Laser Arrangement - Air-Cooled Laser, Example
Optical
rail
Water-cooled
Argon laser or
HeCd laser
HeNe
(Laser 2)
Air-cooled
Argon
(Laser 1)
Top view
7470030A
Mirror Alignment Procedure
Align the mirrors when changing to a different laser or application requiring a different laser.
WARNING Risk of personal injury. Direct eye contact with the output beam from any laser may cause
serious damage and possible blindness. Wear laser safety glasses that protect against the wavelengths
being used. Read the information under Heading 1.2, LASER SAFETY PRECAUTIONS before aligning the
mirror.
WARNING Risk of personal injury. If operating a laser with ultraviolet (UV) wavelengths, note that the beam
is invisible to the eye and dangerous to the skin. Wear laser safety glasses that protect against the UV
wavelengths of the laser. Special Fluorescent cards can be used to spot-check the beam position.
2-2
PN 4237470AA
AIR-COOLED LASERS
GENERAL INFORMATION
1
Without turning on the lasers, start up
the system as directed in Chapter 3,
STARTUP, in the ALTRA flow
cytometer Operator's Guide.
2
Open the covers to the sensing area.
a.
Lift up the front cover.
b.
Pull up the interlock rod.
c.
Squeeze the interlock cover release
button.
b
a
DA
NG
ER
d.
PN 4237470AA
d
c
Remove the interlock cover.
3
Remove the two screws and open the
internal laser cover.
4
Defeat the mechanical interlock by
inserting the interlock defeat bar:
a.
Pull the interlock rod up.
b.
Insert the defeat bar.
c.
Push the interlock rod down.
2-3
2
AIR-COOLED LASERS
GENERAL INFORMATION
5
6
Gain access to the flow cell:
a.
Pull the deflection plates straight
out.
b.
Loosen the thumbscrew holding
the beam-shaping lens assembly in
place.
c.
Remove the beam-shaping lens
assembly.
Thumbscrew
H.V.
ON
Beam-shaping
lens assembly
Deflection
plates
Remove the flow cell tip.
H.V.
ON
2-4
PN 4237470AA
AIR-COOLED LASERS
GENERAL INFORMATION
7
Turn the INTERLOCK OVERRIDE
keyswitch on the Cytometer to defeat
the electrical interlock.
Turn key clockwise
to operate lasers
LASER ON/OFF
8
Remove the forward scatter sensor. Do
not disconnect wires. Let the forward
scatter sensor hang outside the door to
the sort collection area.
9
Remove the pneumatic shutter. Do not
disconnect the tubing.
INTERLOCK OVERRIDE
INTERLOCK OPEN
RESET
MAIN POWER
10 Install the short laser targets in place of
the forward scatter sensor and the
pneumatic shutter. The screws to fasten
the targets should be down.
11 At the touch screen, turn Laser 1 or
Laser 2 (HeNe laser) On. Refer to
Chapter 3, STARTUP, in the ALTRA
flow cytometer Operator's Guide.
PN 4237470AA
2-5
2
AIR-COOLED LASERS
GENERAL INFORMATION
12 Step 13 shows how to adjust the mirror.
Adjust the mirror (reflecting the beam you are aligning) to guide the beam through the
targets as shown.
Note: Always align the laser closest to the flow cell first. Then align each one further
away from the flow cell, in order.
Top View
HeNe
(Laser 2)
Laser
targets
2-6
Laser
beams
633 mirror
reflects 633 nm
and transmits
320 to 365 nm
and 488 nm
Air-cooled
Argon
(Laser 1)
488 mirror
reflects 488 nm
and transmits
320 to 365 nm
HeCd or
Water-cooled
laser
Beam
reducer
UV mirror
reflects 325
to 365 nm
PN 4237470AA
AIR-COOLED LASERS
GENERAL INFORMATION
13 To adjust the mirror:
a.
b.
c.
Loosen the height setscrew and
adjust height so beam strikes
center of mirror.
Loosen the fine adjustment and
rail setscrews. Move the stage
along the optical rail so beam
strikes middle of mirror. Tighten
the fine adjustment setscrew.
Tighten the rail setscrew so the
mirror and post do not rock on the
rail. Keep the setscrew loose
enough so that the fine adjustment
knob can still move the mirror and
post along the rail.
d.
Loosen height setscrew and rotate
mirror so beam goes to first target.
Tighten height setscrew.
e.
Use the horizontal and vertical
rotation knobs to guide the beam
through the first target. Look for
laser light to appear on the second
target.
f.
If the light on the second target is
a wide circle of light to the left side
of the hole (away from the PMTs),
use the fine adjustment knob to
move the post and mirror further
back along the rail. Repeat step e.
g.
If the light on the second target is
a tight point, adjust vertical and
horizontal rotation to get beam
through second target.
h.
Tighten the rail setscrew.
Vertical
rotation
knob
Horizontal
rotation
knob
Fine
adjustment
knob
Height
setscrew
Fine
adjustment
setscrew
Rail
setscrew
14 Close the shutter on the laser.
PN 4237470AA
2-7
2
AIR-COOLED LASERS
GENERAL INFORMATION
15 To align a second air-cooled laser,
repeat steps 11 to 14 for that laser.
16 Close the shutter on that laser. Turn the
flathead screw above that laser aperture
to close the laser shutter.
17 Follow steps 17 to 28 to align the HeCd
laser.
a.
b.
Loosen the height setscrew and
remove the beam reducer that is
positioned between the laser head
and the dichroic alignment mirror.
Turn the HeCd laser on as directed
under Startup in Heading 2.2,
AIR-COOLED ARGON LASERS.
Beam
reducer
Height
setscrew
2-8
PN 4237470AA
AIR-COOLED LASERS
GENERAL INFORMATION
18 Adjust the UV mirror (reflecting the beam) to guide the beam through the targets. (You
need to place a fluorescent target, such as a white business card, in front of the beam to
detect laser action.) Repeat steps 12 and 13 with the UV mirror.
Top View
HeNe
(Laser 2)
Laser
targets
Laser
beams
633 mirror
reflects 633 nm
and transmits
320 to 365 nm
and 488 nm
Air-cooled
Argon
(Laser 1)
488 mirror
reflects 488 nm
and transmits
320 to 365 nm
HeCd or
Water-cooled
laser
Beam
reducer
UV mirror
reflects 325
to 365 nm
19 Replace the beam reducer/expander
and adjust it so the beam goes through
the beam reducer targets. You can look
from the rear of the system and see the
beam as it goes through the beam
reducer. Remove the beam reducer
targets. Adjust the focus so the beam is
about 1/8 in. in diameter as it leaves the
expander.
20 Close the shutter on the HeCd laser.
PN 4237470AA
2-9
2
AIR-COOLED LASERS
GENERAL INFORMATION
21 Reinstall the flow cell tip and adjust it
for proper back reflection.
Aperture
Laser
beam
Reflected
spot
22 Move the flow cell Z-axis to center the
laser beams on the channel of the
quartz cuvette.
23 Reinstall the beam shaper. Adjust the
beam shaper until the beam spot is
centered on the hole in the target
located in the forward scatter position.
24 Remove the targets and replace the
forward scatter sensor and pneumatic
shutter.
25 Perform a full optical alignment as
described in Chapter 3,
REPLACEMENT/ADJUSTMENT
Procedures, in the Special Procedures
and Troubleshooting manual. Leave all
covers and doors open.
2-10
PN 4237470AA
AIR-COOLED LASERS
GENERAL INFORMATION
26 Run beads.
27 Open the door to the detector compartment and set up the filters as shown below.
PMT4
PMT3
PMT2
PMT1
520 - 530
375 - 390
305 - 325
525 BP
381 BP
PMT5
L
5D
32
L
0D
43
L
0D
55
555 - 795
28 Close the door to the detector
compartment.
PN 4237470AA
2-11
2
AIR-COOLED LASERS
GENERAL INFORMATION
29 Check Red HeNe laser alignment:
2-12
a.
Remove any ND filter from the
forward scatter detector.
b.
Close the shutter on the air-cooled
Argon laser.
c.
Open the shutter on the Red HeNe
laser.
d.
At the Camera screen touch
screen, set up a scope display to
show:
r
Forward scatter peak on the
lower trace.
r
PMT5 peak fluorescence
pulse on the upper trace.
e.
Adjust beam-shaping lens
assembly horizontally for
maximum pulse heights. Adjust
gains and PMT voltages as
necessary.
f.
Close the shutter on the Red HeNe
laser.
PN 4237470AA
AIR-COOLED LASERS
GENERAL INFORMATION
30 Check 488 air cooled alignment
co-linear with the Red HeNe laser:
PN 4237470AA
a.
Open the shutter on the air-cooled
Argon laser.
b.
Adjust the 488 mirror horizontal
rotation for maximum forward
scatter pulse height.
c.
Open the shutter on the Red HeNe
laser.
d.
At the Camera screen touch
screen, set up a scope display to
show:
r
PMT5 peak fluorescence on
the lower trace.
r
PMT2 peak fluorescence on
the upper trace.
e.
Adjust the 488 mirror vertical
rotation so the PMT2 pulses on
the upper trace are right above the
PMT5 pulses on the lower trace.
(Do NOT readjust the
beam-shaping lens assembly.)
f.
Close the shutters on the
air-cooled Argon and HeNe lasers.
2-13
2
AIR-COOLED LASERS
GENERAL INFORMATION
31 Prepare to check HeCd laser alignment:
a.
Loosen the height setscrew and remove the beam reducer (take care not to disturb
any of the settings).
b.
Open the shutter on the HeCd laser.
c.
See the filter configuration below.
PMT4
PMT3
PMT2
PMT1
520 - 530
375 - 390
305 - 325
525 BP
381 BP
PMT5
L
5D
32
L
0D
43
L
0D
55
555 - 795
2-14
PN 4237470AA
AIR-COOLED LASERS
GENERAL INFORMATION
32 Check the HeCd alignment:
a.
PN 4237470AA
At the Camera screen touch
screen, set up a scope display to
show:
r
PMT2 peak fluorescence on
the lower trace.
r
PMT5 peak fluorescence on
the upper trace.
b.
Adjust the UV mirror horizontal
rotation to maximize PMT2 pulse
height.
c.
Replace the beam reducer, and
adjust it for maximum PMT2
pulse height. You may need to
adjust the focus. DO NOT readjust
the UV mirror.
d.
At the Camera screen touch
screen, set up a scope display to
show:
r
PMT5 peak fluorescence on
the lower trace.
r
PMT2 peak fluorescence on
the upper trace.
e.
Open the shutter on the Argon
air-cooled laser.
f.
The signal from the Argon laser is
on the bottom trace. Adjust the
beam reducer so the signal from
the HeCd laser on the upper trace
lines up with signal on the bottom
trace.
g.
Press the ON/OFF button on the
remote control box of the HeCd
laser.
h.
Replace all covers.
2-15
2
AIR-COOLED LASERS
GENERAL INFORMATION
33 At the Cytometer, turn the
INTERLOCK OVERRIDE keyswitch to
enable the interlocks.
r
r
Lasers 1 and 2 (air-cooled Argon
and HeNe lasers) turn off when
the covers are replaced.
Turn key
counter clockwise
LASER ON/OFF
INTERLOCK OVERRIDE
INTERLOCK OPEN
RESET
MAIN POWER
Turn lasers 1 and 2 On at the
touch screen.
34 Restore the original filter configuration
and resume normal operation.
Cleaning and Other Maintenance
Turn the air-cooled laser on for about 1 hour at least once a week if the laser is not to be used
for more than 2 weeks.
Refer to the manufacturer’s manual for the cleaning and maintenance procedures involving
the laser.
2-16
PN 4237470AA
AIR-COOLED LASERS
AIR-COOLED ARGON LASERS
2.2
AIR-COOLED ARGON LASERS
Air-cooled Argon lasers permit a wide range of applications from basic immunofluorescence
to more complex kinetics experiments. Its small size makes the air-cooled Argon laser easy to
combine in multilaser configurations. When used alone or with the air-cooled Red HeNe
laser, the air-cooled Argon laser can be mounted for “straight through” operation and ease of
alignment.
Due to low output power, this laser is not recommended for sense-in-air operation. The
air-cooled Argon laser is suitable for sense-in-quartz applications requiring 488 nm excitation
and may require TEM00 mode.
Electrical Power
r
Operates with household current (standard 110 V outlet)
r
Output power: 15 mW of 488-nm output (99% wavelength purity).
Startup
1
At the ALTRA CYTOMETER touch
screen, select Control.
ALTRA CYTOMETER
FS
High
Volt 750
Int
Gain
1
Peak
Gain
PMT1
PMT2
PMT3
PMT4
PMT5
350
1100
1100
930
0
Fl Comp.
None
10
10
10
10
1
Discrim.
10
10
10
1
Sort
Control
Options
Camera
Control
Graph
Service
Scope
Data Rate 0
Sort
#/Sec
2
At the CONTROL SCREEN touch
screen, in the box under the Laser
heading, toggle to Argon Laser On.
Left
0
Right
0
%
0
0
Count
0
0
Limit
0
0
Sample Flow
50
Dec. Inc.
CONTROL SCREEN
Wavelength Requested Power Current
nm
Power
mW*
Amps
Laser
Mix mode
On/Off
Argon
Laser
HeNe
Laser
Beeper
Off
On
488
16
15
5.8
Shutdown
Valves
Off
*Actual power output
Data Rate 0
Sort
Left
Right
#/Sec
0
0
PN 4237470AA
Power up
Valves
%
0
0
Count
0
0
Limit
0
0
Backflush
Delay
0.5 Sec
Sample Flow
50
Dec
Inc
Push
1.0 Sec
Option
Screen
Backflush
2.0 Sec
Valves
Screen
MAIN
Screen
2-17
2
AIR-COOLED LASERS
AIR-COOLED ARGON LASERS
Shutdown
1
At the ALTRA CYTOMETER touch
screen, select Control.
ALTRA CYTOMETER
FS
High
Volt 750
Int
Gain
1
Peak
Gain
PMT1
PMT2
PMT3
PMT4
PMT5
350
1100
1100
930
0
Fl Comp.
None
10
10
10
10
1
Discrim.
10
10
10
1
Sort
Control
Options
Camera
Control
Graph
Service
Scope
Data Rate 0
Sort
#/Sec
2
At the CONTROL SCREEN touch
screen, in the box under the Laser
heading, toggle to Argon Laser Off.
Left
0
Right
0
%
0
0
Count
0
0
Limit
0
0
Sample Flow
50
Dec. Inc.
CONTROL SCREEN
Wavelength Requested Power Current
nm
Power
mW*
Amps
Laser
Mix mode
On/Off
Argon
Laser
HeNe
Laser
Beeper
Off
Off
488
16
15
5.8
Shutdown
Valves
Off
*Actual power output
Data Rate 0
Sort
Left
Right
#/Sec
0
0
2-18
Power up
Valves
%
0
0
Count
0
0
Limit
0
0
Backflush
Delay
0.5 Sec
Sample Flow
50
Dec
Inc
Push
1.0 Sec
Option
Screen
Backflush
2.0 Sec
Valves
Screen
MAIN
Screen
PN 4237470AA
AIR-COOLED LASERS
RED HeNe LASERS
2.3
RED HeNe LASERS
Used together, the Red HeNe and the air-cooled Argon laser enable the ALTRA flow cytometer
to work with a greater range of probes. Due to the Red HeNe’s small size, the air-cooled Argon
can still be mounted in the straight-through configuration. This gives the ALTRA flow
cytometer the unique feature of having only one beam-steering dichroic for its two-laser
configuration.
The Red HeNe laser is suitable for sense-in-quartz applications requiring 633-nm excitation
and may require TEM00 mode. Due to low output power, this laser is not recommended for
sense-in-air operation. Features include:
r
Economical air-cooled operation
r
Operation in TEM00 mode
r
Compact design
r
No maintenance or daily adjustment.
Electrical Power
r
Operates with household current (standard 110 V outlet)
r
10 mW of 633-nm output
r
Operating voltage: 3,100 ±200 Vdc
r
Operating current: 6.5 ±0.2 mA
Controls and Indicators
All control is through the Cytometer.
Laser Head
The laser (see Figure 2.2) is mounted inside the Cytometer.
Figure 2.2 Laser Head - Red HeNe Laser
T
IA
AD FR
RR D U
LASE ITT E ERT
IS EM S AP
THI
I
O ON
RE M
PN 4237470AA
S
PO
EX
AVOID
UR
E
Laser
shutter
2-19
2
AIR-COOLED LASERS
RED HeNe LASERS
Startup
1
At the ALTRA CYTOMETER touch
screen, select Control.
ALTRA CYTOMETER
FS
High
Volt 750
Int
Gain
1
Peak
Gain
PMT1
PMT2
PMT3
PMT4
PMT5
350
1100
1100
930
0
Fl Comp.
None
10
10
10
10
1
Discrim.
10
10
10
1
Sort
Control
Options
Camera
Control
Graph
Service
Scope
Data Rate 0
Sort
#/Sec
2
At the CONTROL SCREEN touch
screen, in the box under the Laser
heading, toggle to HeNe Laser On.
Left
0
Right
0
%
0
0
Count
0
0
Limit
0
0
Sample Flow
50
Dec. Inc.
CONTROL SCREEN
Wavelength Requested Power Current
nm
Power
mW*
Amps
Laser
Mix mode
On/Off
Argon
Laser
HeNe
Laser
Beeper
Off
On
488
16
15
5.8
Shutdown
Valves
On
*Actual power output
Data Rate 0
Sort
Left
Right
#/Sec
0
0
2-20
Power up
Valves
%
0
0
Count
0
0
Limit
0
0
Backflush
Delay
0.5 Sec
Sample Flow
50
Dec
Inc
Push
1.0 Sec
Option
Screen
Backflush
2.0 Sec
Valves
Screen
MAIN
Screen
PN 4237470AA
AIR-COOLED LASERS
RED HeNe LASERS
Shutdown
1
At the ALTRA CYTOMETER touch
screen, select Control.
ALTRA CYTOMETER
FS
High
Volt 750
Int
Gain
1
Peak
Gain
PMT1
PMT2
PMT3
PMT4
PMT5
350
1100
1100
930
0
Fl Comp.
None
10
10
10
10
1
Discrim.
10
10
10
1
Sort
Control
Options
Camera
Control
Graph
Service
Scope
Data Rate 0
Sort
#/Sec
2
At the CONTROL SCREEN touch
screen, in the box under the Laser
heading, toggle to HeNe Laser Off.
Left
0
Right
0
%
0
0
Count
0
0
Limit
0
0
Sample Flow
50
Dec. Inc.
CONTROL SCREEN
Wavelength Requested Power Current
nm
Power
mW*
Amps
Laser
Mix mode
On/Off
Argon
Laser
HeNe
Laser
Beeper
Off
On
488
16
15
5.8
Shutdown
Valves
Off
*Actual power output
Data Rate 0
Sort
Left
Right
#/Sec
0
0
PN 4237470AA
Power up
Valves
%
0
0
Count
0
0
Limit
0
0
Backflush
Delay
0.5 Sec
Sample Flow
50
Dec
Inc
Push
1.0 Sec
Option
Screen
Backflush
2.0 Sec
Valves
Screen
MAIN
Screen
2-21
2
AIR-COOLED LASERS
GREEN HeNe LASERS
2.4
GREEN HeNe LASERS
Though only a 1.5 mW laser, the Green HeNe can produce data that is indistinguishable from
larger air-cooled or water-cooled lasers.
The Green HeNe uses the same mounting hardware as the Red HeNe. The mounting
hardware must be purchased separately.
The Green HeNe’s 544-nm line can improve signal-to-noise ratios by reducing
auto-fluorescence and raman scatter. Features include:
r
Compact design
r
Operation in TEM00 mode
r
No maintenance or daily adjustments.
Electrical Power
r
Operates on household current (standard 110 V outlet)
r
1.5 mW of 544-nm output power
r
Operating current: 6.5 ±0.2 mA
r
Operating voltage: 3,000 ±100 Vdc.
Controls and Indicators
If the Green HeNe laser is used as the second laser, on/off is through the Cytometer. It is
controlled via the Cytometer control screen under the HeNe laser button.
If used as a third laser, on/off is through the power supply.
Power Supply
When the Green HeNe is used as the third laser, the keyswitch on the power supply (see
Figure 2.3) turns the laser on/off.
2-22
PN 4237470AA
AIR-COOLED LASERS
GREEN HeNe LASERS
Figure 2.3 Power Supply - Green HeNe Laser When Used As a Third Laser
7470018A
If the laser is used as the second laser, the power supply is mounted inside the Cytometer. If
the laser is used as the third laser, the power supply is external to the Cytometer. The laser
head (see Figure 2.4) should always be mounted on the optical bench inside the Cytometer.
Figure 2.4 Laser Head - Green HeNe Laser
05
0
P-
1
99 1994
H BE
M
VE
NO T
478
27
-L
G
MF
S/N
R
R
SE
LA CT H
IS
U
IT
TH ROD S W H
P LIE DR
MP S/C ON E
CO HH IATI NC
D D MA S
RA R RD
RFO DA
J
PE TAN CFD R
S 21
PTE
HA
-C
B
SU
Laser
shutter
7470026A
PN 4237470AA
2-23
2
AIR-COOLED LASERS
GREEN HeNe LASERS
Startup
1
At the ALTRA CYTOMETER touch
screen, select Control.
ALTRA CYTOMETER
FS
High
Volt 750
Int
Gain
1
Peak
Gain
PMT1
PMT2
PMT3
PMT4
PMT5
350
1100
1100
930
0
Fl Comp.
None
10
10
10
10
1
Discrim.
10
10
10
1
Sort
Control
Options
Camera
Control
Graph
Service
Scope
Data Rate 0
Sort
#/Sec
2
At the CONTROL SCREEN touch
screen, in the box under the Laser
heading, toggle to HeNe Laser On.
Left
0
Right
0
%
0
0
Count
0
0
Limit
0
0
Sample Flow
50
Dec. Inc.
CONTROL SCREEN
Wavelength Requested Power Current
nm
Power
mW*
Amps
Laser
Mix mode
On/Off
Argon
Laser
HeNe
Laser
Beeper
Off
On
488
16
15
5.8
Shutdown
Valves
On
*Actual power output
Data Rate 0
Sort
Left
Right
#/Sec
0
0
2-24
Power up
Valves
%
0
0
Count
0
0
Limit
0
0
Backflush
Delay
0.5 Sec
Sample Flow
50
Dec
Inc
Push
1.0 Sec
Option
Screen
Backflush
2.0 Sec
Valves
Screen
MAIN
Screen
PN 4237470AA
AIR-COOLED LASERS
GREEN HeNe LASERS
Shutdown
1
At the ALTRA CYTOMETER touch
screen, select Control.
ALTRA CYTOMETER
FS
High
Volt 750
Int
Gain
1
Peak
Gain
PMT1
PMT2
PMT3
PMT4
PMT5
350
1100
1100
930
0
Fl Comp.
None
10
10
10
10
1
Discrim.
10
10
10
1
Sort
Control
Options
Camera
Control
Graph
Service
Scope
Data Rate 0
Sort
#/Sec
2
At the CONTROL SCREEN touch
screen, in the box under the Laser
heading, toggle to HeNe Laser Off.
Left
0
Right
0
%
0
0
Count
0
0
Limit
0
0
Sample Flow
50
Dec. Inc.
CONTROL SCREEN
Wavelength Requested Power Current
nm
Power
mW*
Amps
Laser
Mix mode
On/Off
Argon
Laser
HeNe
Laser
Beeper
Off
On
488
16
15
5.8
Shutdown
Valves
Off
*Actual power output
Data Rate 0
Sort
Left
Right
#/Sec
0
0
PN 4237470AA
Power up
Valves
%
0
0
Count
0
0
Limit
0
0
Backflush
Delay
0.5 Sec
Sample Flow
50
Dec
Inc
Push
1.0 Sec
Option
Screen
Backflush
2.0 Sec
Valves
Screen
MAIN
Screen
2-25
2
AIR-COOLED LASERS
HeCd LASERS
2.5
HeCd LASERS
The HeCd laser kit has been specifically designed for intracellular Calcium studies. It is not
recommended for DNA applications due to the multi-mode beam profile. Because of the
inherent properties of the HeCd laser, it may not be suitable for applications requiring high
resolution, such as chromosome or DNA ploidy analysis.
The HeCd laser option comprises three products:
r
The Optical Upgrade kit includes the mirrors and optical rail for directing the laser
beams into the sensing area.
r
Mounting feet.
r
An air-cooled Omnichrome 3074-30M helium-cadmium Laser.
This manual briefly covers installation requirements, operation, service and maintenance,
and precautions. For more information, refer to the Omnichrome Helium-Cadmium Laser
Instruction Manual.
The HeCd laser emits only in the UV range at 325 nm. The guaranteed power is 20 mW at
this wavelength, but is typically between 20 and 30 mW. The beam is multimode (not a single
point of light but a combination of several nodes of light). Because of the lasing parameters,
the laser power is not adjustable.
The HeCd laser does not include a power meter. For applications that require tracking of the
laser power, you can purchase a power meter for use in the UV range from the laser
manufacturer or other optical supply companies.
The power output is constant throughout the day after the initial warm-up period. Over the
lifetime of the laser, the power output gradually decreases.
Electrical Power
The HeCd laser is available for operation from either:
r
104-128 Vac or
r
220/240 Vac, 6 A, 50-60 Hz.
The box on top of the power supply is used for 220/240 Vac operation.
The power outlet can be the same as used for an optional Printer. Do not use the circuit used
by the ALTRA flow cytometer.
Refer to the Installation chapter in the ALTRA flow cytometer Reference manual for
additional electrical requirements.
Controls and Indicators
Power Supply
The Power Supply has a Laser Control knob, an Emergency ON/OFF keyswitch and a row of
LED indicators (see Figure 2.5).
2-26
PN 4237470AA
AIR-COOLED LASERS
HeCd LASERS
Figure 2.5 Power Supply - HeCd Lasers
Laser
control
knob
LED
indicators
POWER ON
STANDBY
OPERATE
SHUTDOWN
Cd HEATER
He HEATER
OFF
B
TEMP LOCKOUT
ON
Omnichrome ®
EMERGENCY ONLY
MODEL 100
Emergency
ON/OFF
keyswitch
7470013A
The Laser Control knob selects one of three settings as shown in Table 2.1:
Table 2.1 Power Supply Control Settings - HeCd Lasers
Setting
When to Use
Description
OPERATE
For normal operation.
Lasing usually starts 2 or 3 minutes after setting to OPERATE. In
some cases lasing may start immediately.
STANDBY
When the laser will not be used for
>2 hours, but <4 hours.
Lasing stops about 5 minutes after setting to STANDBY. Warm-up
time is reduced when going from STANDBY to OPERATE.
SHUTDOWN
To shutdown the laser.
Initiates a 3-minute shutdown sequence. Lasing stops about
2.5 min after setting to SHUTDOWN. Power is off after 3 minutes.
Note: The 3-minute shutdown sequence sweeps cadmium vapor
still in plasma tube to a special reservoir near the cathode.
Without the 3-minute shutdown, cadmium vapor condenses on
the inner surfaces of the plasma tube which can reduce the laser
life.
PN 4237470AA
2-27
2
AIR-COOLED LASERS
HeCd LASERS
The LED indicators show the laser states described in Table 2.2.
Table 2.2 Power Supply Indicators - HeCd Lasers
LED Indicator
When Illuminated
Troubleshooting
POWER ON
When power is on
None.
Cd Heater
When in OPERATE
mode
Cadmium in laser is heated to supply cadmium vapor for lasing action. If
you observe constant 1 second dim/bright pulses, call your Service
Representative. The dim/bright cycles indicate a possible problem.
He Heater
When in OPERATE
mode and system is
adjusting helium
pressure
Normally illuminates for about 20 minutes after startup and occasionally
flickers on and off throughout the day. If on continuously throughout the
day, call your Service Representative.
TEMP LOCKOUT Briefly after shutdown,
and anytime laser is too
warm to be started
After shutdown, the system must cool down for 5 to 10 minutes before
restarting. When illuminated, restarting is prevented until the laser cools
down.
Laser Head
The laser emission POWER indicator lights anytime lasing is possible (see Figure 2.6). The
laser beam is invisible. Avoid the beam path when the emission indicator is lit.
Figure 2.6 Laser Head - HeCd Lasers
A
Emission
POWER
indicator
POWER
B
Laser
shutter
(Closed)
C
DANGER
7470011A
2-28
PN 4237470AA
AIR-COOLED LASERS
HeCd LASERS
Startup
Unlike the other air-cooled lasers on the ALTRA flow cytometer, the HeCd laser is controlled
from its power supply. Except for the INTERLOCK OVERRIDE keyswitch, settings at the
workstation or the Cytometer do not affect the HeCd laser.
Note: When the laser will not be used for more than 2 weeks, start the laser and let it run for
about 1 hour every week.
Procedure
1
Start the ALTRA flow cytometer as
directed in the Chapter 3, STARTUP, in
the ALTRA flow cytometer Operator's
Guide.
2
At the HeCd laser power supply, turn
the Emergency ON/OFF keyswitch to
ON.
3
Turn the Laser Control knob to
OPERATE.
POWER ON
STANDBY
OPERATE
SHUTDOWN
Cd HEATER
He HEATER
OFF
TEMP LOCKOUT
ON
EMERGENCY ONLY
4
PN 4237470AA
B
Omnichrome
®
MODEL100
After 2 to 3 minutes, the laser emits the
laser beam. Let the laser warm up for
15 minutes before running samples.
2-29
2
AIR-COOLED LASERS
HeCd LASERS
Shutdown
The normal shutdown procedure uses the SHUTDOWN setting on the Laser Control knob
(refer to Table 2.1). This timed shutdown continues plasma discharge until all cadmium
plasma in the bore condenses in a special area near the cathode. The Emergency ON/OFF
keyswitch is for emergency purposes only as directed by the following CAUTION message.
CAUTION Risk of damage to the instrument. If you stop lasing immediately by using the ON/OFF keyswitch,
cadmium vapor condenses on the inner surfaces of the plasma tube and this can reduce the laser life. The
ON/OFF keyswitch immediately stops lasing by stopping plasma discharge in the laser plasma tube. Do not
use the ON/OFF keyswitch for shutdown of the laser EXCEPT in an emergency.
1
Turn the Laser Control knob to
SHUTDOWN.
POWER ON
STANDBY
OPERATE
SHUTDOWN
Cd HEATER
He HEATER
OFF
ON
EMERGENCY ONLY
2-30
2
After about 2.5 minutes, the laser stops
lasing. After about 3 minutes, power is
off to the laser.
3
Shut down the rest of the system as
described in Chapter 8, SHUTDOWN,
in the ALTRA flow cytometer
Operator's Guide.
B
TEMP LOCKOUT
Omnichrome
®
MODEL100
PN 4237470AA
3WATER-COOLED LASERS 3
3.1
GENERAL INFORMATION
The following water-cooled lasers are available as options to the ALTRA flow cytometer:
r
Coherent Innova Enterprise II ion laser system (hereafter referred to as the Enterprise II
series lasers)
r
Coherent Innova 300 Series ion laser system (hereafter referred to as the 300 series
lasers)
r
Coherent Innova 70 Spectrum ion laser system (hereafter referred to as the Spectrum
laser).
The information under this section (Heading 3.1, GENERAL INFORMATION) is generic and
applies to all the water-cooled lasers listed above. Information specific to each laser listed
above is included in a separate section in this chapter.
Installation Requirements
Do not unpack or install the option yourself. The option must be unpacked and installed by a
Beckman Coulter Representative. Certain internal adjustments of the lasers and the
instrument may be required at the time of installation.
Refer to the Installation chapter in the ALTRA flow cytometer Reference manual for other
installation requirements.
Heat and Ventilation
The ALTRA flow cytometer dissipates approximately 2,400 W (8,191 Btu/hour) of heat.
Depending upon the type and power of the associated lasers, the total heat dissipation can
increase considerably. Be sure to consider this to ensure that the room where the ALTRA flow
cytometer is located is properly ventilated and can remove the generated heat.
For example: an water-cooled Enterprise II series laser dissipates about 7,500 W
(25,590 Btu/hour) of heat. Note that the Enterprise II laser is a special case where the heat
can be removed into the surrounding room with an optional chiller or down the drain with
the cooling water. See the laser manufacturer’s specifications for details.
The system must be operated in an air-conditioned space. Keep room temperature between
18°C and 29°C (64°F and 85°F).
Refer to the Installation chapter in the ALTRA flow cytometer Reference manual for other
installation requirements.
Operation
The water-cooled lasers have numerous advanced operating features. The information in this
chapter explains the minimum necessary for operation with the ALTRA flow cytometer. For
complete operating instructions, refer to the specific laser manufacturer’s manual.
Changing Laser Wavelength
Refer to the laser manual to change the laser wavelength. Remember to also set the
wavelength at the remote control box to maintain power meter calibration.
PN 4237470AA
3-1
WATER-COOLED LASERS
GENERAL INFORMATION
When working with jet-in-air flow cells, other changes may be necessary. Contact your
Beckman Coulter Representative for more information.
Aligning the Mirrors
To align the mirrors you need to be aware of the arrangement of the lasers, the path the laser
beam follows and the filter configuration. This configuration changes depending on the mix
of air-cooled vs. water-cooled lasers installed on your instrument.
With the addition of lasers, the orientation of the lasers are changed. All lasers point toward
the right side of the instrument into an optical rail supporting three dichroic alignment
mirrors, one for each laser (see Figure 3.1). The dichroic alignment mirrors reflect the laser
light into the sensing area.
Figure 3.1 Laser Arrangement - Water-Cooled Lasers in a Three Laser Configuration
Optical
rail
Water-cooled
Argon laser or
HeCd laser
HeNe
(Laser 2)
Air-cooled
Argon
(Laser 1)
Top view
7470030A
The water-cooled lasers can be set up for either 488-nm operation, or UV operation (refer to
the laser manufacturer’s manual to switch the laser back and forth). For 488-nm operation,
switch the dichoric mirror from the air-cooled Argon laser to the water-cooled laser. The
air-cooled Argon laser is not used when the water-cooled laser is operated at 488 nm.
Mirror Alignment Procedure
Align the mirrors when changing to a different laser or application requiring a different laser.
WARNING Risk of personal injury. Direct eye contact with the output beam from any laser will cause
serious damage and possible blindness. Wear laser safety glasses that protect against the wavelengths
being used. Read the information under Heading 1.2, LASER SAFETY PRECAUTIONS before aligning the
mirror.
3-2
PN 4237470AA
WATER-COOLED LASERS
GENERAL INFORMATION
WARNING Risk of personal injury. If operating a laser with ultraviolet (UV) wavelengths, note that the beam
is invisible to the eye and dangerous to the skin. Wear laser safety glasses that protect against the UV
wavelengths of the laser. Special Fluorescent cards can be used to spot-check the beam position.
1
Without turning on the lasers, start up
the system as directed in Chapter 3,
STARTUP, in the ALTRA flow
cytometer Operator's Guide.
2
Open the covers to the sensing area.
a.
Lift up the front cover.
b.
Pull up the interlock rod.
c.
Squeeze the interlock cover release
button.
b
a
DA
NG
ER
d.
PN 4237470AA
d
c
Remove the interlock cover.
3
Remove the two screws and open the
internal laser cover.
4
Defeat the mechanical interlock by
inserting the interlock defeat bar:
a.
Pull the interlock rod up.
b.
Insert the defeat bar.
c.
Push the interlock rod down.
3-3
3
WATER-COOLED LASERS
GENERAL INFORMATION
WARNING Risk of electrical shock. The high
voltage carried in the deflection plates can
shock. Do NOT touch the deflection plates if the
H.V. ON button is lit. If the H.V. ON button is lit,
press the button to turn the high voltage off.
5
Check the H.V. ON button. If it is lit,
press the button to turn off the high
voltage to the deflection plates.
6
Gain access to the flow cell:
7
3-4
a.
Pull the deflection plates straight
out.
b.
Loosen the thumbscrew holding
the beam-shaping lens assembly in
place.
c.
Remove the beam-shaping lens
assembly.
Thumbscrew
H.V.
ON
Beam-shaping
lens assembly
Deflection
plates
Remove the flow cell.
PN 4237470AA
WATER-COOLED LASERS
GENERAL INFORMATION
8
Turn the INTERLOCK OVERRIDE
keyswitch on the Cytometer to defeat
the electrical interlock.
Turn key clockwise
to operate lasers
LASER ON/OFF
9
INTERLOCK OVERRIDE
INTERLOCK OPEN
RESET
MAIN POWER
Remove the forward scatter sensor. Do
not disconnect wires. Let the forward
scatter sensor hang outside the door to
the sort collection area.
10 Remove the pneumatic shutter. Do not
disconnect the tubing.
11 Install the short laser targets in place of
the forward scatter sensor and the
pneumatic shutter. Screws to fasten
targets should be down.
12 Turn on the water-cooled lasers on your
instrument. Allow 30 minutes of warm
up time for the lasers.
13 Close the shutter on one of the
water-cooled lasers while aligning the
second laser.
PN 4237470AA
3-5
3
WATER-COOLED LASERS
GENERAL INFORMATION
14 Align the water-cooled laser by adjusting the dichroic alignment mirror reflecting the
laser beam to guide it through the targets.
Note: If the laser is 488 nm wavelength this will be a 488 nm mirror.
Top View
HeNe
(Laser 2)
Laser
targets
3-6
Laser
beams
633 mirror
reflects 633 nm
and transmits
320 to 365 nm
and 488 nm
Argon
(Laser 1)
488 mirror
reflects 488 nm
and transmits
320 to 365 nm
Water-cooled
argon laser
Beam
reducer
UV mirror
reflects 325
to 365 nm
PN 4237470AA
WATER-COOLED LASERS
GENERAL INFORMATION
15 To align the mirror:
a.
b.
c.
Loosen the height setscrew and
adjust height so the beam strikes
the center of mirror.
Loosen the fine adjustment and
rail setscrews. Move the stage
along the optical rail so the beam
strikes the middle of the mirror.
Tighten the fine adjustment
setscrew.
Tighten the rail setscrew so the
mirror and post do not rock on the
rail. Keep the setscrew loose
enough so that the fine adjustment
knob can still move the mirror and
post along the rail.
d.
Loosen the height setscrew and
rotate the mirror so the beam goes
to the first target. Tighten the
height setscrew.
e.
Use the horizontal and vertical
rotation knobs to guide the beam
through the first target. Look for
laser light to appear on the second
target.
f.
If the light on the second target is
a wide circle of light, use the fine
adjustment knob to move the post
and mirror further back along the
rail. Repeat step e.
g.
If the light on the second target is
a tight point, adjust the vertical
and horizontal rotation to get the
beam through the second target.
h.
Tighten the rail setscrew.
Vertical
rotation
knob
Horizontal
rotation
knob
Fine
adjustment
knob
Height
setscrew
Fine
adjustment
setscrew
Rail
setscrew
16 Close the shutter on the laser.
PN 4237470AA
3-7
3
WATER-COOLED LASERS
GENERAL INFORMATION
17 Open the shutter on a second
water-cooled laser, if present, to
perform an alignment on that one.
To align a second water-cooled laser,
repeat steps 14 and 15.
18 Align a water-cooled laser with a beam
reducer if you are using the air-cooled
and water-cooled lasers together. The
beam reducer is used to reduce the
diameter of the water-cooled laser
beam to equal that of the air-cooled
beam.
Beam
reducer
Height
setscrew
3-8
PN 4237470AA
WATER-COOLED LASERS
GENERAL INFORMATION
19 The example uses a water-cooled UV
laser line and the air-cooled Argon
laser.
PN 4237470AA
a.
Loosen the height setscrew and
remove the beam reducer.
b.
Turn the water-cooled laser on as
directed in the Startup instructions
for the appropriate laser listed in
this chapter.
c.
Adjust the UV mirror (reflecting
the beam) to guide the beam
through the targets (see step 14).
(You need to place a fluorescent
target, such as a white business
card, in front of the beam to detect
the laser action.) Repeat steps 14
and 15 with the UV mirror.
d.
Replace the beam reducer and
adjust it so the beam goes through
the targets. You can look from the
rear of the system and see the
beam as it goes through the beam
reducer. Adjust the focus so the
beam as it leaves the expander is
about an eighth of an inch in
diameter. Just before the 633
mirror, the water-cooled laser
beam is about 1/16 in. in diameter.
e.
Close the shutter on the
water-cooled laser.
f.
Remove the targets and replace the
forward scatter sensor and
pneumatic shutter.
g.
Perform a full optical alignment as
described in the Special
Procedures and Troubleshooting
manual. Leave all covers and doors
open.
h.
Run beads.
3-9
3
WATER-COOLED LASERS
GENERAL INFORMATION
20 Open the door to the detector compartment and set up the filters as shown below.
PMT4
PMT3
PMT2
PMT1
520 - 530
375 - 390
305 - 325
525 BP
381 BP
PMT5
L
5D
32
L
0D
43
L
0D
55
555 - 795
21 Continue the alignment.
Close the door to the detector
compartment.
22 Turn on the air-cooled Argon laser at
the Cytometer control screen and close
the shutter.
23 Open the shutter on the water-cooled
laser.
3-10
PN 4237470AA
WATER-COOLED LASERS
GENERAL INFORMATION
24 At the Camera screen touch screen, set
up a scope display to show:
r
Forward scatter peak fluorescence
on the lower trace.
r
PMT2 peak fluorescence on the
upper trace.
25 Adjust beam-shaping lens assembly
horizontally for maximum pulse
heights.
26 Adjust gains and PMT voltages as
necessary.
27 Close the shutter on the water-cooled
laser.
28 Open the shutter on the air-cooled
Argon laser.
29 Adjust the 488 mirror horizontal
rotation for maximum forward scatter
pulse height.
30 Open the shutter on the water-cooled
laser.
PN 4237470AA
3-11
3
WATER-COOLED LASERS
GENERAL INFORMATION
31 At the Camera screen touch screen, set
up a scope display to show:
r
PMT5 peak fluorescence on the
lower trace (from the air-cooled
Argon laser).
r
PMT2 peak fluorescence on the
upper trace (from the UV
water-cooled laser).
32 Adjust the UV mirror vertical rotation
so the PMT2 pulses on the upper trace
are right above the PMT5 pulses on the
lower trace.
Note: DO NOT readjust the
beam-shaping lens assembly.
33 Close the shutters on the air-cooled
Argon and water-cooled lasers.
34 Adjust the UV mirror horizontal
rotation to maximize PMT2 pulse
height.
35 Replace the beam reducer, and adjust it
for maximum PMT2 pulse height. You
many need to adjust the focus.
Note: DO NOT readjust the UV mirror.
3-12
PN 4237470AA
WATER-COOLED LASERS
GENERAL INFORMATION
36 The signal from the UV water-cooled
laser is on the upper trace. Adjust the
beam reducer so the signal from the
water-cooled laser on the upper trace
lines up with signal on the bottom
trace.
37 Press the ON/OFF button on the
remote control box of the water-cooled
laser.
38 Replace all covers.
39 At the Cytometer, turn the
INTERLOCK OVVERRIDE keyswitch
to enable the interlocks.
r
r
Lasers 1 and 2 (air-cooled Argon
and HeNe lasers) turn off when
the covers are replaced.
Turn key
counter clockwise
LASER ON/OFF
INTERLOCK OVERRIDE
INTERLOCK OPEN
RESET
MAIN POWER
Turn lasers 1 and 2 On at the
touch screen.
40 Restore the original filter configuration
and resume normal operation.
Cleaning and Other Maintenance
Startup the water-cooled laser for about 1 hour at least once a week if the laser will not be
used for more than 2 weeks.
Refer to the manufacturer’s manual for the cleaning and maintenance procedures involving
the laser.
PN 4237470AA
3-13
3
WATER-COOLED LASERS
ENTERPRISE II SERIES LASERS
3.2
ENTERPRISE II SERIES LASERS
The Enterprise II Series laser option contains three products:
r
ENTERPRISE Laser with dual 488 nm and UV capability.
r
ENTERPRISE Optics Kit
r
t
Beam steering optics
t
ND filter assembly (splits 488 nm and UV laser lines).
ENTERPRISE Monitoring Hardware
The manufacturer’s manual is the Coherent Innova Enterprise II Operator's manual. The
manufacturer’s manual briefly covers installation requirements, operation, service,
maintenance and precautions. Read the manufacturer’s manual before operating the laser.
Floor Loading and Space
The total system weight including this laser head and power supply is:
567 kg (1,260 lb).
The dimensions of the laser head and power supply are:
Component
Width
Height
Length
Laser Head
18.5 cm (7.3 in.)
17.3 cm (6.8 in.)
76.2 cm (30.0 in.)
Laser Power Supply
43.9 cm (17.3 in.)
19.3 cm (7.6 in.)
50.3 cm (19.8 in.)
Electrical Power
r
r
3-14
Input power:
t
Single-phase with ground, 50 or 60 Hz
t
208 to 240 Vac ±10%, 7.0 Κva maximum.
Maximum input current vs. Input voltage:
t
34 A (rms) @ 208 V (rms)
t
31 A (rms) @ 230 V (rms)
t
29 A (rms) @ 240 V (rms).
PN 4237470AA
WATER-COOLED LASERS
ENTERPRISE II SERIES LASERS
Cooling Water
CAUTION Risk of damage to instrument. If the water temperature falls below the ambient dew point, water
can condense inside the power supply and can cause catastrophic damage. Do not let the water
temperature fall below the ambient dew point.
Table 3.1 Cooling Water Specifications - Enterprise II Series Lasers
Item
Specification
Flow rate, minimum
8 L (2 U.S. gal.) per minute
Flow rate, maximum
16 L (4.0 U.S. gal.) per minute
Incoming water pressure
1.4 to 4.2 Kg/cm2 (20 to 60 psi)
Incoming water temperature
10 to 45°C (50 to 110°F
Heat load to cooling water
4.5 kW (256 Btu/minute)
Heat load to air at power supply
300 W maximum
Controls and Indicators
Power Supply
The power supply has a voltage range of 190 to 260 Vac, single-phase 50/60 Hz input. The
power supply is air-cooled, high-frequency switching and has front panel LEDs (see Table 3.2
and Figure 3.2).
Table 3.2 Power Supply Controls/Indicators - Enterprise II Series Lasers
Control/Indicators
Description
MAIN POWER indicator
When lit, indicates the power is on.
LASER EMISSION indicator
When lit, indicates the system is running.
CURRENT LIMIT indicator
When lit, indicates that the maximum allowable current has been
reached.
When flashing, indicates the need to replace power supply, remote
control module, or RS-232 device.
SYSTEM FAULT indicator
When lit, indicates a heat exchanger fault.
When flashing, indicates the power supply is over temperature.
HEAD FAULT indicator
When lit, indicates the power supply is defective.
When flashing, indicates system is taking an automatic gas fill.
INTERLOCK FAULT indicator
When lit, indicates an open interlock fault exists.
When flashing, indicates an interlock fault occurred but the fault is no
longer present.
LASER POWER keyswitch
PN 4237470AA
Key control for turning the power supply on/off.
3-15
3
WATER-COOLED LASERS
ENTERPRISE II SERIES LASERS
Figure 3.2 Power Supply - Enterprise II Series Lasers
7470019A
Remote Control Module
The remote control module provides complete system control. All system information
including operating status, control settings, and diagnostics, appears on a 4-line,
20-character, back-lit LCD display (see Figure 3.3 and Table 3.3).
Figure 3.3 Remote Control Module - Enterprise II Series Lasers
7470020A
3-16
PN 4237470AA
WATER-COOLED LASERS
ENTERPRISE II SERIES LASERS
Table 3.3 Remote Control Module - Enterprise II Series Lasers
Control
Description
POWER
Increases or decreases the primary and secondary power output.
ON/OFF
Turns the laser on and off.
SELECT
Allows access to the Main Menu and selects an item from the menu.
EXIT
Returns to the previous menu.
SCROLL
Scrolls to the next or previous menu options or changes a value in a parameter selection
menu.
Laser Head
The Enterprise II Series laser head features (see Figure 3.4):
r
A water-cooled design
r
A sealed-mirror, Innova plasma tube
r
An integral SuperInvar resonator
r
Automated mirror alignment.
Figure 3.4 Laser Head - Enterprise II Series
LASER EMISSION
indicator
7470024A
Aperture
control
PN 4237470AA
3-17
3
WATER-COOLED LASERS
ENTERPRISE II SERIES LASERS
Startup
Note: Start the water-cooled Argon laser and leave it powered on for about 1 hour at least
once a week if the laser is not to be used for more than 2 weeks.
3-18
1
Start the ALTRA flow cytometer as
directed in the Chapter 3, STARTUP, in
the ALTRA flow cytometer Operator's
Guide.
2
Turn on the cooling water for the laser.
3
Turn the keyswitch on the power
supply to ON.
4
Turn the Main power switch ON.
PN 4237470AA
WATER-COOLED LASERS
ENTERPRISE II SERIES LASERS
Set Laser Power
1
Use the arrow keys on the remote
control module to set the power (50 to
200 mW at 488 nm).
Shutdown
PN 4237470AA
1
Press the ON/OFF button on the
remote control module.
2
Turn the keyswitch on the power
supply to OFF.
3-19
3
WATER-COOLED LASERS
300 SERIES LASERS
3.3
3
Let cooling water continue to flow until
at least 5 minutes after the laser is
turned off.
4
Follow the directions under Chapter 8,
SHUTDOWN, in the ALTRA flow
cytometer Operator's Guide to shut
down the rest of the system.
300 SERIES LASERS
The 300 Series laser option contains three products:
r
The Optical Upgrade kit which includes the mirrors and optical rail for directing the
laser beams into the sensing area
r
Mounting feet
r
A 300 Series laser (5-W all lines) with ultraviolet (UV) capability.
The manufacturer’s manual is the Coherent Innova 300 Series Ion Laser Operator's manual.
The manufacturer’s manual briefly covers installation requirements, operation, service,
maintenance and precautions. Read the manufacturer’s manual before operating the laser.
Floor Loading and Space
The total system weight including this laser head and power supply is:
576 kg (1,280 lb).
The dimensions of the system are
3-20
Component
Width
Height
Length
Laser Head
15.0 cm (5.9 in.)
14.7 cm (5.8 in.)
122.7 cm (48.3 in.)
Laser Power Supply
43.2 cm (17.0 in.)
17.8 cm (7.0 in.)
53.4 cm (21.0 in.)
PN 4237470AA
WATER-COOLED LASERS
300 SERIES LASERS
Electrical Power
The 300 Series lasers require:
r
Input power: 3-phase with ground, no neutral
r
Voltage: 208 Vac ±10%, 50 or 60 Hz
r
Current:
t
45 A/phase @ 208 Vac
t
55 A/phase @ 208 Vac
t
60 A/phase @ 208 Vac.
The electrical service for the laser should have a main power disconnect switch located in the
same room as the laser.
Refer to the Installation chapter in the ALTRA flow cytometer Reference manual for
additional electrical requirements.
Cooling Water
CAUTION Risk of damage to instrument. If the water temperature falls below the ambient dew point, water
can condense inside the power supply and can cause catastrophic damage. Do not let the water
temperature fall below the ambient dew point.
Table 3.4 Cooling Water Specifications - 300 Series Lasers
Item
Specification
Flow rate, minimum
8.5 L (2.2 U.S. gal.) per minute
Flow rate, maximum
11.6 L (3.0 U.S. gal.) per minute
Maximum static pressure
90 psi
Pressure drop (inlet - outlet)
22 psi to 40 psi
Water temperature
10 to 35°C (50 to 95°F)
Resistivity
5.0 kW-cm to 2.0 MW-cm
pH
6 to 8
Hardness
<100 mg/L (5.9 grains/gal.) or 100 ppm of calcium
Controls and Indicators
Power Supply
The only controls on the power supply are the ON/OFF keyswitch and a series of LED
indicators (see Figure 3.5). All indicators light when the keyswitch is in the ON position.
Interlocks on the power supply are described in the manufacturer’s manual.
PN 4237470AA
3-21
3
WATER-COOLED LASERS
300 SERIES LASERS
Figure 3.5 Power Supply - 300 Series Lasers
®
COHERENT® INNOVA 300
OFF
HK POWER
+15V
-15V
+15V CPU
CPU
ON
±15V CPU
7470031A
Remote Control Module
The remote control module (see Figure 3.6) can only be used when the keyswitch on the
power supply is in the ON position. See Figure 3.6 and Table 3.5 for the controls and their
functions.
Figure 3.6 Remote Control Module - 300 Series Lasers
COHERENT
INNOVA 300
LASER
EMISSION
MEMORY
MENU
SELECT
SET
ON/OFF
TUNE
PWR TRK
LIGHT
7470012A
3-22
PN 4237470AA
WATER-COOLED LASERS
300 SERIES LASERS
Table 3.5 Remote Control Module - 300 Series Lasers
Control
Description
ON/OFF
Turns laser on/off.
Scroll Arrows
r With laser in light regulation, scroll arrows increase/decrease laser power.
r With laser in current regulation, scroll arrows increase/decrease laser current.
r After pressing MENU, scroll arrows toggle through submenus, or settings on submenu.
MENU
r Pressing at default display to display submenu titles.
r Press at a Menu display to display a previous menu level, or return to default display.
SELECT
After pressing MENU, press SELECT to select submenu on display.
SET
r Pressing SET at default display initiates a 15-second PowerTrack® calibration routine.
r On a submenu display of various settings, press to choose a particular setting.
Refer to laser manual for more instructions.
PWR TRK
Press to activate PowerTrack mode. PT ON appears on display. If PowerTrack active, PWR TRK LED
lights.
Refer to manufacturer’s manual for more instructions.
LIGHT
Press to toggle between light and current regulation. Light regulation is active when LIGHT LED lights.
Display also indicates regulation mode.
MEMORY
Refer to manufacturer’s manual for instructions.
Laser Head
Unlike the other two lasers, the 300 Series lasers (see Figure 3.7) are controlled from a remote
control box. Settings at the workstation or the Cytometer do not affect the 300 Series lasers.
Interlocks on the laser head are described in the manufacturer’s manual.
PN 4237470AA
3-23
3
WATER-COOLED LASERS
300 SERIES LASERS
Figure 3.7 Laser Head - 300 Series Lasers
Laser
Shutter
Laser
emission
indicator
Aperture
control
Vertical
search tab
Horizontal
rotation high
reflector
Vertical
rotation high
reflector
High
reflector
assembly
7470014A
Startup
Note: Start the water-cooled Argon laser on for about 1 hour at least once a week if the laser
will not be used for more than 2 weeks.
3-24
1
Start the ALTRA flow cytometer as
directed in the Chapter 3, STARTUP, in
the ALTRA flow cytometer Operator's
Guide.
2
Turn on the cooling water for the laser.
PN 4237470AA
WATER-COOLED LASERS
300 SERIES LASERS
3
4
At the laser power supply, turn the
keyswitch to ON. All LEDs on the front
panel of the power supply light. If they
do not all light, refer to the laser
manufacturer’s manual.
®
COHERENT INNOVA 300
®
OFF
HK POWER
+15V
-15V
+15V CPU
CPU
ON
±15V CPU
At the remote control box, press the
ON/OFF button.
COHERENT
INNOVA 300
LASER
EMISSION
PN 4237470AA
5
After a 30-second start delay, the laser
emits the laser beam.
6
At the remote control box, press the
LIGHT button to put the laser in light
regulation. In light regulation, the laser
system increases or decreases the
current to achieve the power setting.
MEMORY
MENU
SELECT
SET
ON/OFF
TUNE
PWR TRK
LIGHT
3-25
3
WATER-COOLED LASERS
300 SERIES LASERS
7
If PT ON does not display on the
remote control box, press the PWR
TRK button.
IMPORTANT Risk of unpredictable results. Auto
Light Cal uses brief laser output oscillations to
calibrate the PowerTrack feature. These
oscillations can cause unpredictable results
from the ALTRA flow cytometer. Only use Auto
Light Cal when turning the laser on. Be sure
Auto Light Cal is off when running samples.
8
9
Turn Auto Light Cal on:
a.
Press the MENU button.
b.
Press the scroll arrows until Auto
Light Cal appears on the display.
c.
Press the SELECT button until Yes
appears.
d.
Press the SET button.
e.
Press the MENU button.
Let the laser warm up for about
5 minutes.
10 Turn Auto Light Cal off.
3-26
PN 4237470AA
WATER-COOLED LASERS
300 SERIES LASERS
Set Aperture
The 300 Series lasers must operate in TEM00 mode. The TEM mode depends on the specific
plasma tube, laser power, wavelength and aperture setting. Refer to the laser manufacturer’s
manual to determine the TEM mode.
Generally, aperture settings between 4 and 8 give TEM00 mode at about 500 mW at 488 nm.
You can often see if the laser is in some other TEM mode by looking at peak fluorescence
pulses of fluorospheres. If the peak pulse has two peaks or a shoulder, the laser may not be in
TEM00 mode.
Set Laser Power
Press the up or down Scroll Arrows on the remote control box (refer to Figure 3.6) to set the
laser power. At 488 nm, the suggested operating power is between 200 mW and 500 mW.
Tune Laser for Maximum Power
Because of the PowerTrack feature, the 300 Series lasers do not need tuning on a daily basis.
After startup, the PowerTrack feature keeps the laser tuned until the first time you close the
laser shutter or change the power. After that, PowerTrack does not actively tune the laser
until the next startup.
Tune the laser once a week according to the following procedure:
PN 4237470AA
1
Press the TUNE button on the remote
control module.
2
Adjust the horizontal high reflector
control on the laser head for maximum
power as indicated by the laser power
bar graph on the remote control box.
3
Adjust the vertical high reflector
control for maximum power.
4
Press the TUNE button on the remote
control module. The laser returns to its
previous settings.
3-27
3
WATER-COOLED LASERS
SPECTRUM LASER
Shutdown
1
Press the ON/OFF button on the
remote control module.
COHERENT
INNOVA 300
LASER
EMISSION
2
Turn the keyswitch on the power
supply to the OFF position.
MEMORY
MENU
SELECT
SET
ON/OFF
TUNE
PWR TRK
LIGHT
®
COHERENT INNOVA 300
®
OFF
HK POWER
3.4
3
Let cooling water continue to flow until
at least 5 minutes after the laser is
turned off.
4
Follow the directions under Chapter 8,
SHUTDOWN, in the ALTRA flow
cytometer Operator's Guide to shut
down the rest of the system.
+15V
-15V
+15V CPU
CPU
±15V CPU
ON
SPECTRUM LASER
The Spectrum laser has these features:
3-28
r
Mixed gas Argon/Krypton laser
r
Multiwavelength operation including a 647 nm laser line
PN 4237470AA
WATER-COOLED LASERS
SPECTRUM LASER
The Spectrum laser option contains two products:
r
I-70 Spectrum laser (2.5 W multiline, 250 W, 647 nm)
r
Mounting hardware.
The manufacturer’s manual is the Coherent Innova I-70 Spectrum Operator's manual. The
manufacturer’s manual briefly covers installation requirements, operation, service,
maintenance and precautions. Read the manufacturer’s manual before operating the laser.
Floor Loading and Space
The total system weight including this laser head and power supply is:
567 kg (1,260 lb).
The dimensions of the system are:
Component
Width
Height
Length
Laser Head
18.5 cm (7.3 in.)
16.5 cm (6.5 in.)
113.8 cm (44.8 in.)
Laser Power Supply
44.7 cm (17.6 in.) 22.4 cm (8.8 in.)
36.6 cm (14.4 in.)
Electrical Power
The Spectrum laser requires:
r
Input power: 3-phase with ground
r
Voltage: 208 ±10% Vac 50 or 60 Hz
r
40 A per phase @ 208 Vac.
Cooling Water
CAUTION Risk of damage to instrument. If the water temperature falls below the ambient dew point, water
can condense inside the power supply and can cause catastrophic damage. Do not let the water
temperature fall below the ambient dew point.
Table 3.6 Cooling Water Specifications - Spectrum Laser
Item
Specification
Flow rate
8.5 L (2.2 U.S. gal.) per minute
Incoming water pressure
1.41 to 4.23 Kg/cm2 (20 to 60 psi)
Incoming water temperature
10 to 35°C (50 to 95°F)
Controls and Indicators
Power Supply
The power supply can be operated from the power supply (see Figure 3.8) or the remote
control module (see Figure 3.9). The remote control module is connected to the power
supply by a cable and provides you with full control of the laser from a convenient location. It
supports:
PN 4237470AA
3-29
3
WATER-COOLED LASERS
SPECTRUM LASER
r
User selectable light and current regulation through the remote control module (see
Figure 3.9 and Table 3.7).
r
Continuous tube pressure monitoring to alert you to the need for a gas fill (see
Figure 3.9 and Table 3.7).
r
Continuous water flow monitoring - if the water flow drops below the minimum levels,
the power supply shuts off to protect the system (see Figures 3.8 and 3.9).
r
Safety interlocks to prevent laser operation in an unsafe condition (see Figure 3.9 and
Table 3.7).
Figure 3.8 Power Supply - Spectrum Laser
7470017A
Table 3.7 Power Supply and Remote Control Module Controls and Indicators - Spectrum Laser
Control/Indicator
Location
Description
LASER EMISSION indicator
Power supply
Illuminates when the power supply is turned ON.
LASER indicator
Remote control
module
WATER FLOW indicator
Power supply
WATER indicator
Remote control
module
SYSTEM ON/OFF pushbutton Power supply
SYSTEM ON/OFF keyswitch
3-30
Red LED that lights continuously when water flow drops below
2.2 gal./minute and flashes when laser tube pressure is too low.
Energizes power supply.
Remote control
module
PN 4237470AA
WATER-COOLED LASERS
SPECTRUM LASER
Table 3.7 Power Supply and Remote Control Module Controls and Indicators - Spectrum Laser
Control/Indicator
Location
Description
INTERLOCK indicator
Remote control
module
LED on indicates an open interlock. Check the laser head cover, power
supply cover, or fuse cover.
FAULT indicator
Remote control
module
LED on indicates:
r No ac power, or
r Tube under pressure, or
r Tube over current, or
r ± 15 V power supply failure.
Regulation mode switch
Remote control
module
Four-position switch for selecting current control (CUR) or UV or one
of three light regulation ranges (LO, MED, HI).
POWER control
Remote control
module
Adjusts tube current from minimum to maximum when in current
regulation (CUR) mode.
START pushbutton
Remote control
module
Initiates the start sequence
Analog readout
Remote control
module
Analog readout can be set to show either LIGHT POWER, TUBE
CURRENT, or TUBE VOLTAGE.
Remote Control Module
The remote control module (see Figure 3.9 and Table 3.7) houses:
r
The status indicators
r
A main power keyswitch
r
An analog readout showing the tube current, tube voltage and output power.
Figure 3.9 Remote Control Module - Spectrum Laser
7470016A
PN 4237470AA
3-31
3
WATER-COOLED LASERS
SPECTRUM LASER
Laser Head
Figure 3.10 Laser Head - Spectrum Laser
Vertical
scanning
laser
Beam
shutter
7470021A
3-32
PN 4237470AA
WATER-COOLED LASERS
SPECTRUM LASER
Startup
Note: Start the water-cooled Argon laser on for about 1 hour at least once a week if the laser
will not be used for more than 2 weeks.
PN 4237470AA
1
Start the ALTRA flow cytometer as
directed in the Chapter 3, STARTUP, in
the ALTRA flow cytometer Operator's
Guide.
2
Turn on the cooling water for the laser.
3
At the laser power supply, turn the
keyswitch to ON. The WATER FLOW
LED lights momentarily while the
water flow is verified.
4
Press the ON button. The LASER
EMISSION indicator lights.
5
After a 30-second start delay, the laser
emits the laser beam.
3-33
3
WATER-COOLED LASERS
SPECTRUM LASER
6
If the beam shutter is not already open,
set the beam shutter control knob so
that the white line is parallel to the
tube bore to open the shutter.
7
Set the regulation mode to CUR and
turn the POWER knob to full power.
Set Laser Power
1
3-34
Adjust the POWER knob on the remote
control module to the desired power
level.
PN 4237470AA
WATER-COOLED LASERS
SPECTRUM LASER
Tune Laser for Maximum Power
1
Adjust the vertical and horizontal
adjustment knobs to tune the laser
power to Maximum on the remote
control module.
Shutdown
PN 4237470AA
1
Turn the interlock key on the remote
control module counterclockwise to
turn off the laser.
2
Wait 5 minutes after turning off the
laser before turning off the water.
3-35
3
WATER-COOLED LASERS
SPECTRUM LASER
3-36
PN 4237470AA
4STORAGE OPTIONS 4
4.1
GENERAL INFORMATION
The following storage media are available as options for the ALTRA flow cytometer:
r
Iomega® Zip™ Drive (hereafter referred to as the Zip drive).
r
Yamaha Recordable CD-ROM (hereafter referred to as the CD-ROM).
r
Iomega jaz® internal drive (hereafter referred to as the jaz drive).
The information under this section (Heading 4.1, GENERAL INFORMATION) is generic and
applies to all the storage media listed above. Information specific to each storage media listed
above is included in a separate section in this chapter. Refer to the manufacturer’s manual for
each option for operating instructions.
Magnetic Storage Media Care
Despite quality-control standards, some magnetic storage media become defective. To help
prevent data loss, adhere to the following guidelines:
4.2
r
Always keep backup copies of your data on separate diskettes or other magnetic storage
media.
r
Do not touch recording surfaces.
r
Keep media away from magnetic fields, such as those generated around radios and
phones.
r
Do not bend diskettes, CD-ROMs or other storage media.
r
Always store magnetic storage media at room temperature.
Zip™ DRIVE
The Zip drive is an internal IDE storage device. A starter set of storage media is included in
the kit and additional disk packs are available through Beckman Coulter, or at most computer
supply stores.
The Iomega Zip Drive kit includes:
4.3
r
Iomega Zip Drive
r
1 pack of media (250 MB each)
RECORDABLE CD-ROM DRIVE
The Yamaha Recordable CD-ROM is an internal “one-time recordable,” many-times readable,
media allowing you to archive and retrieve Histogram and Listmode files to CD-ROM media,
for long-term storage. It is intended for use with the FlowCentre multimedia workstation.
The CD-ROM kit contains:
r
The Yamaha internal CD-ROM Drive
r
10 Pak of CD-ROM media (650 MB each)
We recommend that you purchase an Uninterruptable Power Supply locally for the
Workstation and Recordable CD-ROM drive. Any power interruptions while you are
recording to a CD renders that CD-ROM disk completely useless.
PN 4237470AA
4-1
STORAGE OPTIONS
jaz® DRIVE
The Recordable CD-ROM Drive includes two different software packages to create CD-ROMs.
The software packages are Adaptec Easy CD-Creator Deluxe Edition and Adaptec Direc-CD.
Easy CD Creator
Easy CD-Creator creates CD-ROMs that can be read in Windows 98, Windows 95,
Windows 3.1, Windows NT™, and MS-DOS® operating system, thus allowing for full
cross-platform compatibility. Files archived to CD-ROM using Easy CD-Creator however can
be read in XL™ System II™ or Elite™ Software. Easy CD-Creator also allows for recording of
audio, video and graphics presentations to CD-ROM as well as Multi-Session recording.
CD-Creator also features a CD Copier program, that duplicates CD-ROMs.
Note: It is a violation of copyright laws to duplicate any software from CD-ROMs which is
copyright protected. The customer assumes full responsibility for any such action. Beckman
Coulter assumes no liability whatsoever.
Easy CD-Creator Installation Procedures
1.
Locate the Easy CD-Creator software program kit, and insert the CD-ROM into the drive.
2.
The installation screen appears. Use the mouse cursor and click on EASY CD-Creator to
install the software, following instructions on the screen.
3.
After installation, from the Windows 98 desktop, click on START, PROGRAMS, EASY
CD-CREATOR and select EASY CD-CREATOR DELUXE, to start the recording program.
4.
Place a blank CD-ROM media disk into the caddy supplied with the recordable CD-ROM
drive.
5.
It is recommended that you utilize the EASY CD-CREATOR WIZARD to duplicate a
CD-ROM diskette.
6.
Follow the on-screen instructions to record a test file (any file) onto the blank CD-ROM
media.
r
Files recorded in ISO-9660 format can be read by any operating system, for
example: Windows 98, Windows 95, Windows 3.1, Windows NT, Macintosh®, and
MS-DOS.
r
Files recorded in Joliet format, allow for long file names in Windows 98 and
Windows 95, and are readable in Windows 3.1 and MS-DOS.
7.
Once the test file has been recorded, remove the CD-ROM from the caddy and insert into
the CD-ROM drive present in the workstation.
8.
Open up Windows Explorer and locate the CD-ROM drive in the workstation, and click
on it twice. Verify that the test file recorded is present on the CD-ROM disk.
For additional information on the features of Adaptec Easy CD-Creator, please refer to the
Adaptec’s User Guide.
4.4
jaz® DRIVE
The jaz drive is an internal storage drive. It provides 2-GB of storage per disk.
4-2
PN 4237470AA
5Autoclone™ SORTING OPTION 5
5.1
INTRODUCTION
The EPICS ALTRA Autoclone sorting option is a single-cell deposition system only. It
performs two functions.
1. It is a programmable single-cell deposition system.
r
It sorts a user-defined number of cells per well into a multiwell microculture plate.
r
Automates hybridoma screening for research and development of monoclonal
antibodies, sterile sorting of bacterial cultures, and screening for high-density
monoclonal markers.
r
The system accepts standard 6, 12, 24, 48, 60, or 96 well microculture plates for
automated cell deposition.
2. The second function, Sort Matrix, automates the process of determining the optimum
Drops Delay setting.
r
A holder for microscope slides is used when performing a Sort Matrix.
It depends upon the ALTRA host system for programming instructions, data accumulation,
and electrical power.
When the Autoclone sorting option is not in use, the rear cover and disable knob must be in
place.
5.2
INSTALLATION REQUIREMENTS
r
The installation requirements for the ALTRA system remain unchanged when the
Autoclone system is installed. Refer to the Installation chapter in the Reference manual
for ALTRA system installation requirements.
r
Do not unpack or install the Autoclone system yourself. The system must be unpacked
and installed by your Beckman Coulter Representative.
Installation Calibration Procedure
After initial installation or reinstallation of the ALTRA software, perform this abbreviated
calibration procedure:
PN 4237470AA
5-1
Autoclone™ SORTING OPTION
INSTALLATION REQUIREMENTS
IMPORTANT Risk of electric shock. The
voltage on the deflection plates can shock you.
DO NOT touch the deflection plates if the H.V.
ON button is lit. If the button is lit, press the
button to turn the deflection plates off.
5-2
1
If the H.V. ON button is lit, press the
button to turn off the deflection plates.
2
Remove the rear plate covering the
Autoclone arm by turning the disable
knob counterclockwise.
PN 4237470AA
Autoclone™ SORTING OPTION
INSTALLATION REQUIREMENTS
3
At the cytometer MAIN touch screen,
select in order:
Options
Autoclone
Control
4
At the Autoclone screen, select:
Adjust
Screen
5
At the Autoclone Adjustment screen,
select:
Calibrate
6
7
PN 4237470AA
After calibration, retract the Autoclone
arm. Select:
At the Autoclone Control screen,
select:
Autoclone
Control
Retract
Screen
5-3
5
Autoclone™ SORTING OPTION
STARTUP
8
Follow the instructions on the Retract
screen and press:
OK
9
5.3
Replace the Autoclone cover.
STARTUP
1
If the HPSS option is NOT installed,
check the pressure gauges on the
compressor. If the system vacuum or
pressure is outside limits, adjust them
as described in the Special Procedures
and Troubleshooting manual.
External
compressor
-15
-10
-20
-5
-25
-30
SYSTEM VACUUM
30
20
10
40
50
60
SYSTEM PRESSURE
5-4
PN 4237470AA
Autoclone™ SORTING OPTION
STARTUP
IMPORTANT Risk of electric shock. The voltage
on the deflection plates can shock you. DO NOT
touch the deflection plates if the H.V. ON button is
lit. If the H.V. ON button is lit, press the H.V. ON
button to turn the deflection plates off.
PN 4237470AA
2
If the H.V. ON button is lit, press the
H.V. ON button to turn off the
deflection plates.
3
Remove the rear plate covering the
Autoclone sorting option arm by
turning the disable knob
counterclockwise.
5-5
5
Autoclone™ SORTING OPTION
STARTUP
4
At the cytometer MAIN touch screen,
select in order:
Options
Autoclone
Control
Load
.
The Autoclone sorting option arm
extends out to accept the plate holder
and perform autocalibration.
5
Install the multiwell plate holder with
cutout area facing front.
For sort matrix, install the slide adapter
in the plate holder with the microscope
slide to the left.
Slide/plate
holder
For sorting, slide a multiwell plate onto
plate holder so that well A1 is to the
left.
6
5-6
Bend the guides on the plate holder so
that the multiwell plate fits snug.
PN 4237470AA
Autoclone™ SORTING OPTION
STARTUP
WARNING Risk of electric shock. The voltage
on the deflection plates can shock you. DO NOT
touch the deflection plates if the H.V. ON button
is lit. If the H.V. ON button is lit, press the H.V.
ON button to turn the deflection plates off.
7
Enable sheath and setup sort streams as
shown in Chapter 7, SORTING, in the
Operator’s Guide.
8
Turn the STREAM knob until the left
sort stream is flowing straight down.
9
Toggle deflection high voltage off and
on to check the stream position.
Sort
stream
Slide
adapter
Slide/
plate
holder
With high voltage on, the left sort
stream should look the same as the
sheath stream does when high voltage
is off.
PN 4237470AA
5-7
5
Autoclone™ SORTING OPTION
SORT MATRIX
5.4
SORT MATRIX
The Autoclone system performs an
automated Sort Matrix to help determine
correct Drops Delay.
5-8
1
Use the camera cursors to find the
approximate Drops Delay. Refer to
Chapter 7, SORTING, in the Operator’s
Guide.
2
Ensure the Sort Control screen and Sort
Settings box contain correct values.
3
Run a sample of Flow-Check
fluorospheres, or equivalent, and place
a sort region around the fluorospheres.
PN 4237470AA
Autoclone™ SORTING OPTION
SORT MATRIX
4
Set up the sort equation.
Screen tt Sort.
Region name from the region list.
5
Id label in the Autoclone Equation
box
6
PN 4237470AA
Type a name for the sort equation and
press Û.
5-9
5
Autoclone™ SORTING OPTION
SORT MATRIX
7
Select Sort Matrix.
Screen tt Autoclone.
Select Plate.
Up or down arrow until Sort
Matrix is highlighted.
Okay.
8
Enter Slide Characteristics.
Orifice Diameter.
Type 76 or 100 and press Û.
9
Enter Sort Settings
The Sort Settings are received from the
Sort screen, however a Drops Delay
Step value needs to be entered.
Drop Dely.
To accept the current value press Û.
Type a Drops Delay Step value and
press Û.
5-10
PN 4237470AA
Autoclone™ SORTING OPTION
SORT MATRIX
10 Examples of Drops Delay Step values:
A Drops Delay Step of 1 programs the
system to sort a range of 4 drop delays
higher and lower than the Drop Dely in
the sort settings box.
11 Program the slide.
Id label from equation list.
PN 4237470AA
5-11
5
Autoclone™ SORTING OPTION
SORT MATRIX
12 The Id label automatically displays
above the slide next to Eq:.
13 Type the number to be sorted and press
Û.
14 Type a description and press Û.
5-12
PN 4237470AA
Autoclone™ SORTING OPTION
SORT MATRIX
15 Insert a microscope slide in the slide
adapter. Install the slide adapter in the
plate holder with the microscope slide
to the left.
Slide
Slide
adapter
Plate
holder
IMPORTANT Risk of electric shock. The voltage
on the deflection plates can shock you. DO NOT
touch the deflection plates if the H.V. ON button is
lit. If the H.V. ON button is lit, press the H.V. ON
button to turn the deflection plates off.
16 If the H.V. ON button is lit, press the
H.V. ON button to turn off the
deflection plates.
17
PN 4237470AA
Activate.
5-13
5
Autoclone™ SORTING OPTION
SORT MATRIX
18 Examine the slide with a fluorescence
microscope to determine the correct
drop delay value.
Sort Settings to toggle to Send.
Drop Dely.
Type the correct drop delay and press
Û.
19 Save the Autoclone and sort settings
with the protocol.
Screen tt Protocol.
File tt Save.
To use the same name for the protocol
press Û,
OR
Type a name for the protocol and press
Û.
5-14
PN 4237470AA
Autoclone™ SORTING OPTION
SORTING
5.5
SORTING
1
Perform Startup, Heading 5.3, to
prepare the Autoclone system for
sorting.
2
Run a sort matrix, refer to Heading 5.4,
SORT MATRIX, to determine the
correct drop delay for sorting.
3
Run a sample and create or edit the
Gate and Sort regions. Refer to Chapter
5, RUN SAMPLES in the Operator’s
Guide, for instructions on creating and
editing regions.
4
Set up the Autoclone equation.
Screen tt Sort.
Region name from the region list.
PN 4237470AA
5-15
5
Autoclone™ SORTING OPTION
SORTING
5
Id label in the Autoclone Equation
box.
6
Type a name for the equation and press
Û.
7
Select a multiwell plate.
Screen tt Autoclone.
Select Plate.
Up or down arrow until Sort
Matrix is highlighted.
Okay.
5-16
PN 4237470AA
Autoclone™ SORTING OPTION
SORTING
8
Enter Plate Characteristics.
Orifice Diameter.
9
Type 76 or 100 and press Û.
10 Well Timeout
Type the number of seconds to wait for
sorted particles before moving to the
next well position and press Û.
11 Liquid in well
Type in the amount of media in the
wells and press Û.
PN 4237470AA
5-17
5
Autoclone™ SORTING OPTION
SORTING
12 Sort Settings
The sort settings are the same values as
in the sort screen.
The sort settings are saved with the
protocol after performing a Sort Matrix.
See Heading 5.4, SORT MATRIX.
For the Autoclone sorting option, use
the AccuSort™ Mode (Drops 7,
Coincidence Abort = ON).
13 Program the plate with Autoclone
equations.
Id number from the program
equation box.
5-18
PN 4237470AA
Autoclone™ SORTING OPTION
SORTING
14
Well that you want to sort into.
Type the number of cells to be sorted
and press Û.
Type a description and press Û.
The Autoclone equation ID number
displays in the well.
15 To program an entire row at a time.
Id number from the program
equation box.
Number next to the row of wells
you want to sort into.
PN 4237470AA
5-19
5
Autoclone™ SORTING OPTION
SORTING
16 To program an entire column at a time.
Id number from the program
equation box.
Letter under the column of wells
you want to sort into.
17 To program an entire plate at a time.
Id number from the program
equation box.
Fill Plate button.
WARNING Risk of biohazardous
contamination. Keep sample stage area and
sort collection area doors closed during sample
acquisition. Aerosols may be created which
may contain biohazardous materials.
18 Ensure H.V. ON button is lit and
Activate.
5-20
PN 4237470AA
Autoclone™ SORTING OPTION
CALIBRATION
5.6
CALIBRATION
When Calibrate is selected in the
Autoclone Control menu, the system
performs automatic calibration of the
Autoclone sorting option arm movement.
AUTOCLONE SCREEN
96 WELLS
r
Calibrate the Autoclone sorting
option whenever the cytometer
software is reloaded or replaced, or
the CYTO.HEX file is transferred.
r
Lx
Cw
Fine tune the calibration whenever
the stream is not centered in the plate
wells.
Before you Fine Tune the calibration you
must:
ROW
Markers
Hg
r
12 ROW
Retract
Screen
8 COL
COL
CURRENT
Autoclone
Waste
On
SELECTED
Autoclone Cover Off
Inc ROW
Inc COL
Home
Dec ROW
Dec COL
Load
Sort
Screen
Options
Screen
MAIN
Screen
MOVE 13 SELECTED LOCATION
low cell
orizontal
Adjust
Screen
Flow cell
vertical
Beam
vertica
Make sure the system is optically
aligned. Refer to the Special
Procedures and Troubleshooting
manual.
H.V.
ON
Beam
horizontal
PN 4237470AA
5-21
5
Autoclone™ SORTING OPTION
CALIBRATION
r
Use Sort Test to get tight sort streams.
Refer to Chapter 7, SORTING, in the
Operator’s Guide.
r
Toggle Right Sort Enable OFF.
r
Turn the STREAM knob until the left
sort stream is flowing straight down.
Toggle Deflection High Voltage off
and on to check the stream position.
With high voltage on, the left sort
stream should look the same as the
sheath stream does when high voltage
is off.
r
Sort
stream
Slide
adapter
Slide/
plate
holder
Leave Sort Test on.
Fine Tune Calibration
1
5-22
Slide a 96-well plate onto the plate
holder.
PN 4237470AA
Autoclone™ SORTING OPTION
CALIBRATION
2
At the cytometer MAIN menu select
Options
PN 4237470AA
and
Autoclone
Control .
3
Select Inc ROW and Inc COL until 12-D is
displayed to the right of SELECTED.
4
Select MOVE TO SELECTED LOCATION.
5
Toggle Autoclone Waste Off.
6
Select Inc ROW and Inc COL until 1-D is
displayed to the right of SELECTED.
7
Select MOVE TO SELECTED LOCATION.
5-23
5
Autoclone™ SORTING OPTION
CALIBRATION
Adjust
Screen
8
Select
9
Look at the line of sheath fluid on the
plate and use CW and CCW to align the
plate so that the line of sheath fluid is
parallel to the multiwell columns.
.
10 Repeat steps 2 to 9 until the plate is
correctly aligned.
11 Use FWD and BCK buttons to center the
stream in well 1-D.
12 Select
Autoclone
Control and toggle
Autoclone Waste On.
13 Select
5-24
Adjust
Screen
.
PN 4237470AA
Autoclone™ SORTING OPTION
CALIBRATION
14 Select Inc ROW and Inc COL until 12-H is
displayed to the right of SELECTED.
15 Select MOVE TO SELECTED LOCATION.
16 Select
Autoclone
Control and toggle
Autoclone Waste Off.
17 Select
Adjust
Screen
.
18 Use LFT and RGT buttons to center the
stream in well 12-H.
Note: If you cannot get the stream
centered in both wells (1-D and 12-H)
at the same time, repeat steps 11 to 18
to optimize the position of the stream
in both wells.
PN 4237470AA
5-25
5
Autoclone™ SORTING OPTION
CALIBRATION
Adjust Autoclone Sorting Option Speed
You should adjust the speed of the
Autoclone movement if:
r
The Autoclone sorting option is noisy
when it moves.
r
The Autoclone sorting option is
missing wells when it sorts.
The nominal speed values are shown in
the illustration to the right
Noisy Movement
At the cytometer Main touch screen select:
1
2
Autoclone
Control and
Select
X,Y
▲
Adjust
Screen
.
to increase the
speed of left, right, forward, and back
movement.
3
Select
ROT
▲
to increase the
speed of rotation.
5-26
PN 4237470AA
Autoclone™ SORTING OPTION
CALIBRATION
Missing Wells
At the cytometer Main touch screen select:
1
2
Autoclone
Control and
Select
X,Y
▼
Adjust
Screen
.
to decrease the
speed of left, right, forward, and back
movement.
3
Select
ROT
▼
to decrease the
speed of rotation.
PN 4237470AA
5-27
5
Autoclone™ SORTING OPTION
CALIBRATION
5-28
PN 4237470AA
6GATED AMP OPTION 6
6.1
WHAT THIS CHAPTER EXPLAINS
This chapter explains the features of the GATED AMP option for dual laser applications and
imaging optics alignment on your EPICS ALTRA flow cytometer.
6.2
FEATURES OF THE GATED AMP
Time-Domain Gating
With time-domain gating, the ALTRA system is set up with spatially-separated laser beams.
Analyzed particles pass through two interrogation points generating two sets of signals, a few
microseconds apart. Any detector receives signals as the particle passes through both laser
beams.
Time-domain gating allows you to select either signal while blocking other signals received by
the detector. Or you can select both signals from the same detector while still blocking noise
generated in the interval between the two signals.
Signals from the upper laser beam are delayed electronically to synchronize them with the
undelayed signals from the lower laser beam. This delay allows subtraction to be applied to
signals from the upper and lower laser without affecting the ADC functions.
Lower Beam Lockout
Lower beam lockout is a software controlled function that prevents the system from creating a
second gate in acquisition before the first gate is closed. Do not use this feature when sorting
because it may reduce your purity.
Dual Laser Applications
Example: DAPI/FITC for Viable DNA and Immunofluorescence
WARNING Risk of personal injury. This laser operates in the UV (ultraviolet) wavelengths which can injure
you when exposed and which are invisible to the human eye. It can cause eye damage if viewed either
directly or indirectly from reflective surfaces (such as a mirror or a shiny metal surface, including tools). To
prevent eye damage, avoid direct exposure to the beam.
The UV excited DAPI emission spectrum overlaps the 488 nm excited FITC emission
spectrum. By using spatially separated laser beams and the Gated Amp you can separate these
signals. The same detector can be used to collect a delayed FITC and two undelayed DAPI
signals. These signals are amplified separately allowing doublet exclusion by peak-width
comparison.
Example: Calcium Flux with Indo-1 and Two-Color Cell-Surface Markers
Using the 488 nm laser and a HeCd air-cooled UV laser you can measure both FITC and
INDO LONG on the same detector. All the signals from the 488 nm laser are delayed to
synchronize with the signals from the UV laser.
PN 4237470AA
6-1
GATED AMP OPTION
IMAGING OPTICS ALIGNMENT
Example: Hoechst/Chromomycin for Dual-Color Chromosome Sorting
While Hoechst is excited by UV, chromomycin is optimally excited by the 457 nm Argon ion
line. Since both dyes have emission peaks in the blue-green region of the spectrum it is
necessary to use the Gated Amp to resolve the overlap.
Because of energy transfer complications, it is better to take the delayed upper laser signal
from chromomycin-457 nm, and the undelayed lower laser signal from the Hoechst-UV
combination.
Example: Simultaneous Four-Color Clone Studies
When used for four color cell-surface clone markers, the emission spectra of ECD and APC
overlap in the red region. Using spatially separated 633 nm HeNe and 488 nm Argon lasers,
the Gated Amp separates these two signals.
6.3
IMAGING OPTICS ALIGNMENT
The imaging optics module is used to reduce the number of coincidence events detected by a
PMT when a particle is in the first laser at the same time as a triggering event is in the second
laser and the gated amp is open.
6-2
1
Ensure both lasers have been properly
aligned.
2
Open the sensing compartment cover.
PN 4237470AA
GATED AMP OPTION
IMAGING OPTICS ALIGNMENT
3
Lift the curved cover up and out from
between the sample station control
panel and the cytometer touch screen.
Note: This cover contains the HPSS
control panel on units with HPSS
option.
7468188C
4
Unscrew the lock screws and slide off
the PMT and its holder.
PMT
PN 4237470AA
PMT
Lock screw
PMT
holder
PMT holder
Lock screw
6-3
6
GATED AMP OPTION
IMAGING OPTICS ALIGNMENT
5
Slide on the borescope and holder and
tighten with the lock screws.
Borescope
holder
lock screw
Borescope
lock screw
6
Place the focusing screen into the
imaging optics module with the slotted
locknut up. Tighten with a locknut.
Borescope
holder
Borescope
Focusing
screen
Locknut
6-4
7
Place the fiber optic light in the
bandpass slot. Focus the borescope
until there is a sharp view of the
granularity of the focusing template.
8
Remove the focusing screen.
PN 4237470AA
GATED AMP OPTION
IMAGING OPTICS ALIGNMENT
9
Run Flow-Check™ fluorospheres and
adjust the lever on the imaging optics
module to bring the bead flashes into
sharp focus through the borescope.
Note: You might need to adjust the
fluorescence pickup lens lever.
10 Install the pinhole carrier and tighten
with a locknut. Using the knobs, adjust
the pinhole to allow only the desired
laser fluorescence through the pinhole.
Locknut
11 Remove the borescope and holder.
Replace the PMT. Peak up the PMT
signals by pinhole adjustment knobs.
12 Replace and close the covers.
PN 4237470AA
6-5
6
GATED AMP OPTION
SET UP GATED AMP SCREENS
6.4
SET UP GATED AMP SCREENS
The controls and adjustments for the Gated Amp are in two screens on the cytometer, Gated
Amp Assignment and Gated Amp Control. To access these screens, select Options at the
MAIN screen.
Gated Amp Assignment Screen
Figure 6.1 Gated Amp Assignment Screen
c
b
2-6-99
GATED AMP ASSIGNMENT
AUX ASSIGN
AUX-1
AUX-2
SOURCE
PMT-3
SOURCE
PMT-3
PREAMP
GAIN
1.0
PREAMP
GAIN
1.0
SIGNAL
ASSIGN
DELAY ENABLE
FALS
UPP.BM
PMT1
UPP.BM
PMT2
UPP.BM
PMT3
UPP.BM
PMT4
LOW.BM
PMT5
UPP.BM
AUX-1
LOW.BM
AUX-2
LOW.BM
FLUORESC.
COMP.
PRAM A
AUX-1
INT
PRAM B
AUX-2
PEAK
GATED
AMP
ASSIGN
d
SCOPE
SCREEN
OPTION
MAIN
e
7470066C
6-6
PN 4237470AA
GATED AMP OPTION
SET UP GATED AMP SCREENS
Number
Field Name
B
AUX-1 and AUX-2
ASSIGN
Description
r This function allows PMT 2 to 5 to be assigned as an AUX channel.
r To select an AUX source, touch the AUX box affected, then choose from displayed
PMT sources.
r If “None” is selected for AUX-1, PMT4 peak replaces PARAM A everywhere.
r If “None” is selected as AUX-2, there is no PMT5 peak, so nothing will be present.
Example: For FITC and DAPI collection from the same PMT and DAPI collected in
integral and peak for ratio int/peak ratio:
Split beams (Argon 488 upper and UV lower)
Assign PMT3 as source for AUX-1 and AUX-2.
Choose PMT3 to be delayed (UPP.BM)-FITC.
Choose AUX-1 and AUX-2 to be undelayed (LOW.BM) (DAPI).
Assign AUX-1 as integral.
Assign AUX-2 as peak.
c
SIGNAL DELAY
ENABLE/DISABLE
r This function allows the choice of upper (UPP.BM) or lower (LOW.BM) laser/cell
resultant signal to be gated through to their amplifiers for processing.
r Each box displays the current sensor delay status. To change, touch the desired
sensor box.
Example: For FITC and DAPI collection from the same PMT:
Split beams (Argon 488 upper and UV lower)
Assign PMT3 (for DAPI collection) as AUX-1.
Choose PMT3 to be delayed (UPP.BM)-FITC.
Choose AUX-1 to be undelayed (LOW.BM) (DAPI).
d
AUX-1 and AUX-2
PROCESSED
SIGNAL
ASSIGNMENTS
r Choices for AUX-1 are: integral or log. Processed signal choice for AUX-1 is called
PARAM A.
r Choices for AUX-2 are: integral, log, or peak. Processed signal choice for AUX-2 is
called PARAM B.
r To choose the desired processed signal for AUX’s, choose PARAM A or B box, then
choose from the processed signals displayed.
e
AUX-1 and AUX-2
PREAMP GAIN
ADJUST
r Preamp gain can be adjusted from 1.0 to 10.
r This function allows the Aux channel up to a 10X greater signal than the PMT
sourcing it. This is extremely helpful when dealing with a bright and dim marker on
the same cell. By keeping the bright marker’s resultant signal in a nonsaturated mode
and boosting the dim marker’s signal by up to 10X, it is possible to gain a much more
useful signal strength combination.
r This preamp gain selection is previous to the normal AUX-1 int, and the AUX-2 int
and peak gain controls located on the MAIN screen.
r To change, touch PREAMP GAIN affected, then INC or DEC as desired.
PN 4237470AA
6-7
6
GATED AMP OPTION
SET UP GATED AMP SCREENS
Gated Amp Control Screen
Figure 6.2 Gated Amp Control Screen
c
b
1!
2-6-99
GATED AMP CONTROL
1)
WINDOW
CLOCK
5 µSEC
TRIG.
SOURCE
j
FALS
PEAK
SIGNAL
TRIG
DISCRIM.
100
SIGNAL
DELAY
60 µSEC
TRIG.
GAIN
10
LOWER
BM. GATE
LOCKOUT
ENABLE
WIDTH
25 µSEC
i
ENABLE
h
DELAY
55 µSEC
SCOPE
TRACE
SAVE
SETTINGS
RESTORE
SETTINGS
DEFAULT
SETTINGS
FLUORESC.
COMP.
GATED
AMP
ASSIGN
d
SCOPE
SCREEN
OPTION
g
e
MAIN
f
7470065C
Number
Field Name
B
TRIGGER
DISCRIMINATOR
Description
r Discriminates for the trigger source chosen.
r Begins gating process when trigger source is greater in amplitude than discrim.
value.
r (0 - 1000 displayed)
r 0 to 10V
r Touch the box to change the discriminator value.
c
SIGNAL DELAY
r Signal delay chosen should reflect the time between the upper beam and lower beam
peak pulses.
r Selections are: 7 µsec, 20 µsec, 40 µsec, or 60 µsec.
r Current value is shown.
r Touch the box to change signal delay setting.
6-8
PN 4237470AA
GATED AMP OPTION
SET UP GATED AMP SCREENS
Number
Field Name
d
SAVE, RESTORE,
DEFAULT
SETTINGS
Description
r These three functions allow you to: save up to 16 different gated amp setups, recall
up to 16 different gated amp setups, or recall the default settings.
r The functions saved are: trig. source select, clock value, window width and delay, trig.
discrim., trig. gain, signal delay scope trace, selection AUX-1 and AUX-2 sources,
AUX-1 and AUX-2 preamp gains, AUX-1 and AUX-2 int. and peak gains, all sensors
delayed or not status, and all fluorescence settings.
r Touch SAVE SETTINGS or RESTORE SETTINGS, and then choose a letter from the 16
letters that appear.
Note: Existing settings are overwritten if the same letter is selected twice.
r Touch DEFAULT SETTINGS to recall the default settings.
e
LOWER BEAM
TRIGGER-CAUSED
GATE LOCKOUT
ENABLE/DISABLE
r When enabled, this provides the ability to use a sensor excited by both beams as a
trigger source, but only a gate caused by the upper beam.
Note: When enabled, this function decreases sorting purity.
r Current status is shown.
r Touch the box to change the setting.
F
TRACE 1 SCOPE
SELECT
r Selections are: Trig source, Gate window, or Trace 1 preselected on scope screen.
r Current status is shown.
r Touch the box to change the setting.
g
h
DELAY OF GATING
WINDOW
OPENING
r Depends on the clock value.
ENABLE/DISABLE
r Enables or disables the gated amp.
r Maximum = 19X clock, Minimum = 4X clock, Default = 50 µsec.
r Touch the box to change the setting.
r Current status is shown.
i
j
TRIGGER SOURCE
GAIN
r Control is from 0 to 100% of full scale.
TRIGGER SOURCE
r Select a gated amp trigger. The trigger must be caused by the upper beam.
r Touch the box to change the setting.
r Selections are: FALS PEAK, PMT1 PEAK, PMT2 PEAK, PMT3 PEAK, PMT4 PEAK,
PMT5 PEAK, AUX1 PEAK, AUX2 PEAK, FALS LOG, PMT1 LOG, PMT2 LOG, PMT3
LOG, PMT4 LOG, PMT5 LOG, AUX1 LOG, AUX2 LOG.
1)
WIDTH OF GATING
WINDOW
r Depends on the clock value.
r Maximum = 16X clock, Minimum = 1X clock, Default = 25 µsec.
r Touch the box to change the setting.
1!
CLOCK
r Selections are: 0.5 µsec, 1 µsec, 5 µsec (default), 10 µsec.
r Current value is shown.
r Touch the box to change clock setting.
PN 4237470AA
6-9
6
GATED AMP OPTION
SET UP GATED AMP SCREENS
Gated Amp Control Screen Settings
If using
Then set
76 µm, 100 µm or 250 µm quartz SortSense flow cell tip
Clock = 5 µsec
76 µm jet in air flow cell tip
Clock = 0.5 µsec or 1 µsec
70 µm HyPerSortSense quartz flow cell tip
Clock = 5 µsec
76 µm high velocity or HyPerSortSense quartz flow cell tip Clock = 0.5 µsec or 1 µsec
76 µm Quartz flow cell tip
Width between 20 and 30 µsec
100 µm Quartz flow cell tip
Width between 10 and 25 µsec
250 µm Quartz (Profile) flow cell tip
Width between 5 and 10 µsec
76 µm jet in air flow cell tip
Width between 2 and 5 µsec
70 µm HyPerSortSense quartz flow cell tip
Width between 10 and 25 µsec
76 µm high velocity or HyPerSortSense quartz flow cell tip Width between 2 and 5 µsec
76 µm Quartz flow cell tip
Delay between 50 and 60 µsec
100 µm Quartz flow cell tip
Delay between 30 and 40 µsec
250 µm Quartz (Profile) flow cell tip
Delay between 20 and 30 µsec
76 µm jet in air flow cell tip
Delay between 4 and 6 µsec
70 µm HyPerSortSense quartz flow cell tip
Delay between 30 and 40 µsec
76 µm high velocity or HyPerSortSense quartz flow cell tip Delay between 4 and 6 µsec
76vµm Quartz flow cell tip
Signal Delay = 60 µsec
100 µm Quartz flow cell tip
Signal Delay = 40 µsec
250 µm Quartz (Profile) flow cell tip
Signal Delay = 20 µsec
76 µm jet in air flow cell tip
Signal Delay = 7 µsec
70 µm HyPerSortSense quartz flow cell tip
Signal Delay = 7 µsec
76 µm high velocity or HyPerSortSense quartz flow cell tip Signal Delay = 7 µsec
Trigger Discriminator ≈ 100
Any flow cell tip
Trigger Gain ≈10
Lower Bm. Lockout Enabled
(unless sorting, then Disabled)
Fluorescence Compensation Combinations
Subtract
6-10
From
AUX1
PMT1
AUX2
PMT1
PMT1
AUX1
AUX2
AUX1
PN 4237470AA
GATED AMP OPTION
CHECK ALIGNMENT
Subtract
From
PMT1
AUX2
AUX1
AUX2
Software Selectable Amplifier Speed/Sensitivity
Flow Cell Tip
6.5
Amplifier Speed
76 µm or 100 µm SortSense (quartz)
Low
Jet-in-air, BioSense, or HyPerSortSense
High
CHECK ALIGNMENT
1
Turn on and warm up both lasers.
2
Run Flow-Check fluorospheres to
verify correct separation between the
upper and lower beams.
3
Remove the ND1 filter from the
forward scatter detector and cover the
filter slot to keep out room light.
Note: This illustration shows 60 µsec
separation between forward scatter
peak pulses when using 12 psi sheath
pressure and a 76 µm SortSense quartz
flow cell tip.
Scope Display
60 µsec
between
pulses
10 µsec/div
PN 4237470AA
6-11
6
GATED AMP OPTION
CHECK ALIGNMENT
WARNING Risk of personal injury. Reflections
of the laser beam from a shiny object, such as a
screwdriver, or direct viewing of the laser beam
can severely damage your eyes. Pay attention
to the warning labels in the sampling
compartment. Do not wear any jewelry that
might reflect the laser beam. Do not look at the
intersection of the laser beam and the flow
chamber. Use the camera view to observe this
point.
4
Use the 488 nm dichroic mirror vertical
and horizontal adjustments to adjust
laser beam separation.
Vertical
rotation
knob
Horizontal
rotation
knob
Fine
adjustment
knob
Height
setscrew
Fine
adjustment
setscrew
Rail
setscrew
5
Using the dichroic only works when
the laser to be offset is close to the
sampling compartment. The longer the
distance to the compartment, the more
difficult it is to aim the beam properly
into the sampling compartment beam
shaper and achieve proper separation at
the flow cell tip.
If this is the case, it is recommended
that you install beam translators to
offset the beams.
Fluorescence Pickup Lens
1st beam
2nd beam
1st beam aligned
2nd beam not aligned
1st beam
6
6-12
While viewing signals from both
beams, adjust the beam shaper vertical
to the optimal compromise position
indicated by maximizing the second
laser beam signal without much loss on
the first laser beam signal.
2nd beam
Optimal compromise of
dual laser positioning
PN 4237470AA
GATED AMP OPTION
ADJUST WINDOW WIDTH
6.6
ADJUST WINDOW WIDTH
After all the delays have been set up:
Scope Display
1
Trailing edge
of pulse
2
3
PN 4237470AA
Look at the peak pulses for each signal
being collected.
Leading edge
of pulse
Adjust the gate window width so that
the leading or trailing edge of the pulse
is not being clipped.
To display the gate window on the
scope screen, toggle the Trace 1 Scope
Select button on the Gated Amp
Control screen until it displays Gate
Window.
Gate
window
10 µsec/div
6-13
6
GATED AMP OPTION
ADJUST WINDOW WIDTH
6-14
PN 4237470AA
7LARGE PARTICLE SORTING OPTION 7
7.1
DESCRIPTION
The Large Particle Sorting (LPS) option, when installed on the ALTRA Flow Cytometer,
analyzes and sorts large particles (≤200 µm) suspended in media. This option consists of a
large particle sort flow cell, sample tubing assembly, and pressure filter assembly.
The Large Particle Sorting option’s flow cell is sense-in-air (jet-in-air). It has three available
flow cell tip orifice sizes: 200 µm, 280 µm, and 400 µm.
Generally, particles up to one half of the orifice diameter can be sorted with good results.
Larger particles requiring lower pressures and frequencies, consistent with the larger orifice
diameters needed, are best sorted with the Large Particle Sorting Option.
7.2
COMPONENTS
The Large Particle Sorting option consists of four components:
r
Large Particle Sorting Flow Cell and Tips
r
Large Particle Sorting Sample Tubing Assembly
r
Pressure Filter Assembly
r
Beam Shaper, 40 x 80 mm, defocused.
Install all four components before running Large Particle Sorting option samples.
Remove all four components when you are going to run standard or HPSS samples.
Note: Due to the wide range of operating pressures and availability of different sorting
options, be sure to use the appropriate pressure for the corresponding instrument
configuration. Applying high pressure to low-pressure options causes the release of sample
pressure. The Large Particle Sorting Option is not intended for use with high pressure/HPSS
samples. Remove the Large Particle Sorting Option and install the standard flow cell before
running high pressure/HPSS samples.
7.3
INSTALL THE LARGE PARTICLE SORTING OPTION
In normal operation you need to switch between the standard flow cell and the Large Particle
Sorting option flow cell depending upon the type of samples that need to be run in your
laboratory.
Procedure
Follow this procedure to take out the standard flow cell and sample tubing assembly and
install the Large Particle Sorting flow cell, sample tubing assembly, and pressure filter
assembly.
Reverse this procedure when you want to remove the Large Particle Sorting flow cell and
install the standard flow cell.
PN 4237470AA
7-1
LARGE PARTICLE SORTING OPTION
INSTALL THE LARGE PARTICLE SORTING OPTION
1
Press
2
At the ALTRA CYTOMETER touch
screen, press Control tt Shutdown Valves.
3
After the message Valves actions have
completed is displayed, wait 5 minutes
then turn the instrument off as shown
at the right. If the HPSS option is
installed, also turn off the pneumatic
supply.
4
.
LASER ON/OFF
INTERLOCK OPEN
INTERLOCK OVERRIDE
Open the instrument:
a.
Lift the front cover.
b.
Pull up the interlock rod.
c.
Squeeze the interlock cover release
button.
RESET
MAIN POWER
b
a
DA
NG
ER
d.
7-2
d
c
Remove the interlock cover.
PN 4237470AA
LARGE PARTICLE SORTING OPTION
INSTALL THE LARGE PARTICLE SORTING OPTION
WARNING Risk of electrical shock. The high
voltage carried in the deflection plates can
shock. Do NOT touch the deflection plates if the
H.V. ON button is lit. If the H.V. ON button is lit,
press the button to turn the high voltage off.
5
Check the H.V. ON button. If it is lit,
press the button to turn off the high
voltage to the deflection plates.
WARNING If biohazardous materials are
analyzed, decontaminate the system as directed
in the SHUTDOWN chapter in the Operator's
Guide. Wear gloves when handling these parts.
Be especially careful to avoid sticking yourself
with the sharp ends of the sample insertion or
sample pickup rods.
PN 4237470AA
6
Pull the deflection plates straight out.
7
Disconnect the sort pulse alligator clip
from the flow cell body vacuum line.
Thumbscrew
H.V.
ON
Beam-shaping
lens assembly
Deflection
plates
7-3
7
LARGE PARTICLE SORTING OPTION
INSTALL THE LARGE PARTICLE SORTING OPTION
8
Remove the sample tubing assembly
connected to the standard flow cell:
Tubing Brown peek
tubing
connector
a.
Unscrew the tubing connectors at
the flow cell and sample pickup
cap.
b.
Pull the tubing out of the sample
pinch valve.
Pinch
valve
bracket
c.
Save the sample tubing assembly
and the tubing connectors.
Pinch
valve
d.
See the Special Procedures and
Troubleshooting manual for more
detailed instructions.
Orange
peek
tubing
Flow
cell
5
Tubing
connector
9
Remove the standard flow cell from the
instrument:
a.
Unscrew the two screws from the
bimorph.
b.
Pull off the bimorph cable.
Pickup
cap
Bimorph
Knurled
nut
Bimorph
cable
Flow cell
body
Sheath and
vacuum tubing
Screws to
Bimorph
Left side view
7-4
PN 4237470AA
7
LARGE PARTICLE SORTING OPTION
INSTALL THE LARGE PARTICLE SORTING OPTION
10 Disconnect the standard flow cell’s
sheath and vacuum tubing connector:
a.
Press in on the two quick
disconnect fittings.
b.
Pull the tubing connector up and
out.
Vacuum
tubing
Sheath
tubing
Quick
disconnects
11 Install the Large Particle Sorting flow
Alligator
clip
cell body:
a.
Select the size flow cell tip needed
and screw it onto the flow cell
body.
b.
Connect the sort pulse alligator
clip to the flow cell body vacuum
line.
c.
Align the flow cell bracket
connected to the Large Particle
Sorting flow cell body into
position.
d.
Tightly screw in the two saved
screws into the flow cell bracket.
e.
Plug in the bimorph cable.
7
Bracket
12 Open the right side door to the
instrument.
PN 4237470AA
7-5
LARGE PARTICLE SORTING OPTION
INSTALL THE LARGE PARTICLE SORTING OPTION
13 Connect the sheath and vacuum tubing
connector from the Large Particle
Sorting flow cell body to the
instrument:
a.
Align the tubing connector as
shown.
b.
Push down until the connector
snaps into place.
Note: Do not use the tubing connector
if the closed loop tubing is not present
or has an opening. This closed loop
tubing helps to monitor the sheath
pressure.
Closed
loop
tubing
Vacuum
tubing
Sheath
tubing
Quick
disconnects
7470051A
14 Close the right side door to the
instrument.
15 Place the peek tubing end of the Large
Connector
and ferrule
Particle Sorting sample tubing
assembly into the sample pickup cap
and tube:
a.
b.
Pickup
cap
Place a test tube (12 x 75 mm or
12 x 100 mm) on the sample
stage.
Sample
pickup tube
Insert the peek tubing into the
sample pickup cap. Push it in until
the peek tubing touches the
bottom of the sample pickup tube.
Note: Lifting up on the sample
pickup cap makes it easier to
tighten the tubing connector.
c.
Tighten the tubing connector.
Peek tubing
7-6
PN 4237470AA
LARGE PARTICLE SORTING OPTION
INSTALL THE LARGE PARTICLE SORTING OPTION
16 Remove the sample pickup tube.
17 Thread the silicone tubing portion of
the sample tubing assembly through
the sample pinch valve.
Pinch Silicone
valve tubing
Flow
cell
18 Push the end of the sample tubing
assembly all the way onto the sample
insertion rod at the top of the Large
Particle Sorting flow cell body.
Tubing
connector
PN 4237470AA
Sample
pickup
cap
Peek
tubing
7-7
7
LARGE PARTICLE SORTING OPTION
INSTALL THE LARGE PARTICLE SORTING OPTION
19 Install the jet-in-air ring on the pickup
lens, the standard cross forward scatter
obscuration template in front of the
forward scatter detector, and install the
40 x 80 mm defocused beam shaper.
Jet-in-air
ring
STANDARD CROSS
Standard cross
FS obscuration
template
7-8
PN 4237470AA
LARGE PARTICLE SORTING OPTION
INSTALL THE LARGE PARTICLE SORTING OPTION
20 Take off the right side panel of the
instrument.
PN 4237470AA
7-9
7
LARGE PARTICLE SORTING OPTION
INSTALL THE LARGE PARTICLE SORTING OPTION
21 Press the tab on the right
quick-disconnect fitting of the bottom
(sheath air) filter and pull it off.
Note: Some older units without the
HPSS option are configured differently
than pictured and you do not need to
perform steps 20 to 24.
6
9
12
3
15
22 Connect the pressure filter assembly
into the sheath air filter.
23 Position the pressure filter assembly
below the sheath air filter.
Sheath
air filter
Pressure
filter
assembly
Quick
disconnects
24 Close the right side panel of the
instrument.
25 Power up the instrument.
26 At the ALTRA CYTOMETER touch
screen, press Control tt Power Up Valves.
27 Press
and check for leaks at
the flow cell body.
7-10
PN 4237470AA
LARGE PARTICLE SORTING OPTION
INSTALL THE LARGE PARTICLE SORTING OPTION
28 Press
for a few seconds to
flush the tubing.
29 Adjust the Large Particle Sorting flow
cell position by turning the flow cell
stage Z-axis knob until the jetting
stream is centered in the laser beam.
Flow cell
Z-axis
30 Align the optics. See the Special
Procedures and Troubleshooting
manual for instructions.
31 Push the deflection plates back onto
the instrument.
32 Adjust for no side scatter disturbance.
PN 4237470AA
Flow cell
vertical
7-11
7
LARGE PARTICLE SORTING OPTION
REPLACE THE SAMPLE TUBING ASSEMBLY
33 Close the instrument:
7.4
a.
Replace the interlock cover.
b.
Close the front cover.
REPLACE THE SAMPLE TUBING ASSEMBLY
You need to install the Large Particle Sorting option sample tubing assembly whenever you
install the Large Particle Sorting flow cell. Refer to Heading 7.3, INSTALL THE LARGE
PARTICLE SORTING OPTION for instructions.
Replace a defective Large Particle Sorting sample tubing assembly if the instrument
experiences the following only while the Large Particle Sorting option is installed:
r
High background counts.
r
Excessive carryover.
r
Persistent clogging.
Procedure
WARNING Risk of electrical shock. With power running through the instrument, electrical shock can occur.
To protect yourself against electric shock, perform this procedure with the instrument turned off and
disconnected from the power outlets.
7-12
1
Press
2
At the ALTRA CYTOMETER touch
screen, press Control tt Shutdown Valves.
.
PN 4237470AA
LARGE PARTICLE SORTING OPTION
REPLACE THE SAMPLE TUBING ASSEMBLY
3
4
PN 4237470AA
After the message Valves actions have
completed is displayed, wait 5 minutes
then turn the instrument off as shown
at the right. If the HPSS option is
installed, also turn off the pneumatic
supply.
LASER ON/OFF
INTERLOCK OVERRIDE
INTERLOCK OPEN
RESET
MAIN POWER
Open the instrument:
a.
Lift the front cover.
b.
Pull up the interlock rod.
c.
Squeeze the interlock cover release
button.
d.
Remove the interlock cover.
7-13
7
LARGE PARTICLE SORTING OPTION
REPLACE THE SAMPLE TUBING ASSEMBLY
WARNING Risk of biohazardous contamination.
Biohazardous contamination could occur from
contact with biohazardous waste fluid. Avoid
skin contact and dispose of biohazardous waste
in accordance with your local regulations and
acceptable laboratory practices.
5
Remove the sample tubing from the
insertion rod at the top of the Large
Particle Sorting flow cell.
6
Pull the tubing out of the sample pinch
valve.
7
8
Pinch Silicone
valve tubing
Tubing
connector
Sample
pickup
cap
Flow
cell
Peek
tubing
Remove the peek tubing:
a.
Unscrew the tubing connector
from the sample pickup cap.
b.
Pull the peek tubing up and out of
the sample pickup cap.
Perform this step only if you are
replacing defective sample tubing
assembly. Not if you are simply
switching flow cells.
Remove the ferrule and compression
ring from the peek tubing:
9
7-14
a.
Use a screwdriver to twist the
compression ring off the ferrule.
b.
Pull off the ferrule and
compression ring and save them.
Pull off and save the tubing connector.
PN 4237470AA
LARGE PARTICLE SORTING OPTION
REPLACE THE SAMPLE TUBING ASSEMBLY
10 Discard the old sample tubing
assembly.
11 Prepare the new peek tubing:
a.
Slide the saved tubing connector
onto the peek tubing
b.
Slide on the compression ring
(flattened end facing the tubing
connector) and then the ferrule.
Connector
Orange peek
tubing
Note: Be sure to use the sample tubing
assembly for the Large Particle Sorting
option.
Ring
Ferrule
To sample
pickup cap
12 Place the new peek tubing into the
Connector
and ferrule
sample pickup tube:
a.
b.
Place a test tube (12 x 75 mm or
12 x 100 mm) on the sample
stage.
Pickup
cap
Sample
pickup tube
Insert the peek tubing into the
sample pickup cap. Push it in until
the peek tubing touches the
bottom of the sample pickup tube.
Note: Lifting up on the sample
pickup cap makes it easier to
tighten the tubing connector.
Peek tubing
PN 4237470AA
7-15
7
LARGE PARTICLE SORTING OPTION
REPLACE THE SAMPLE TUBING ASSEMBLY
13 Remove the sample pickup tube.
14 Pull the silicone tubing through the
sample pinch valve.
Pinch Silicone
valve tubing
Flow
cell
15 Push the silicone tubing onto the
insertion rod at the top of the Large
Particle Sorting flow cell.
Tubing
connector
Sample
pickup
cap
Peek
tubing
16 Power up the instrument.
17 At the ALTRA CYTOMETER touch
screen, press Control tt Power Up Valves.
18 Press
and check for leaks at
the flow cell body.
7-16
PN 4237470AA
LARGE PARTICLE SORTING OPTION
RUN SAMPLES
19 Press
for a few seconds to
flush the tubing.
20 Close the instrument.
7.5
a.
Replace the interlock cover.
b.
Close the front cover.
RUN SAMPLES
Install Large Particle Sorting option
Install all of the components of the Large Particle Sorting option before running samples.
Refer to Heading 7.3, INSTALL THE LARGE PARTICLE SORTING OPTION.
Select the Flow Cell Tip
There are three sizes of jet-in-air flow cell tips available for the Large Particle Sorting option:
200 µm, 280 µm, and 400 µm.
For best results, select a flow cell tip with an orifice about 10x the size of the cell to be
measured.
Select Settings
Use this chart which contains typical settings for large particle sorting. These values may vary
depending on your configuration.
Table 7.1 Typical Settings for Large Particle Sorting
PN 4237470AA
Flow Cell Tip (Orifice Size)
Frequency (kHz)
Sheath Pressure (psi)
200 µm
8 to 10
5 to 10
280 µm
7.0 to 7.5
6.0 to 7.0
400 µm
2.0 to 2.6
2.0 to 3.0
7-17
7
LARGE PARTICLE SORTING OPTION
RUN SAMPLES
Run Sample
IMPORTANT Incorrect sorting data might be reported if you sort using the Large Particle
Sorting flow cell with the mixer motor on. When sorting with the Large Particle Sorting flow
cell, stop the sample flow before mixing with the mixer motor. After mixing, you can begin
the sample flow again.
Follow the instructions in the Operator’s Guide for running samples and sorting.
7-18
PN 4237470AA
8FORWARD FLUORESCENCE DETECTOR OPTION 8
8.1
DESCRIPTION
The Forward Fluorescence Detector (FFD) option replaces the standard FALS detector when
installed on the instrument. This option detects the range of fluorescence signals that the
standard PMTs 1 through 5 detect while allowing FALS to be concurrently detected.
The FFD also allows finer resolution of submicron particles, through the use of a PMT to
detect the small scatter signals they produce. This option is also useful in the analysis and
sorting of nonspherical cells.
8.2
INSTALL THE FORWARD FLUORESCENCE DETECTOR OPTION
PN 4237470AA
1
Power down the instrument.
2
Open the instrument:
a.
Lift the front cover.
b.
Pull up the interlock rod.
c.
Squeeze the interlock cover release
button.
d.
Remove the interlock cover.
8-1
FORWARD FLUORESCENCE DETECTOR OPTION
INSTALL THE FORWARD FLUORESCENCE DETECTOR OPTION
3
Remove the forward scatter sensor
shield:
a.
Remove the two screws from the
front of the forward scatter sensor
bracket to remove the bottom
shield.
b.
Remove the two screws from the
left of the forward scatter sensor
bracket.
c.
Lift the forward scatter shield off.
Remove
screws
on left
4
Forward
scatter
shield
Remove
screws
in front
Lift the curved cover up and out from
between the sample station control
panel and the cytometer touch screen.
7468188C
8-2
PN 4237470AA
FORWARD FLUORESCENCE DETECTOR OPTION
INSTALL THE FORWARD FLUORESCENCE DETECTOR OPTION
5
Remove the forward scatter sensor.
a.
Use an Allen wrench to remove the
screw from the right of the
bracket.
b.
Slide the forward scatter sensor off
the instrument.
c.
Unplug the bottom left cable
assembly by slowly rocking the
connector back and forth and
pulling down.
d.
Unplug the H.V. ON button wire
connector from the wire harness to
the left of the forward scatter
sensor.
Note: Use the open area from step
4 to access the wire connector.
e.
PN 4237470AA
Set the forward scatter sensor
aside.
Remove
screw
Disconnect
cable
assembly
H.V. ON button
wire connector
8-3
8
FORWARD FLUORESCENCE DETECTOR OPTION
INSTALL THE FORWARD FLUORESCENCE DETECTOR OPTION
6
Plug the bottom left cable assembly
that you disconnected from the forward
scatter sensor into the bottom of the
FFD.
7
Insert the dowel pin on the back of the
FFD into the forward scatter sensor
bracket.
FFD
Screw
8
Plug the H.V. ON button wire
connector from the FFD into the wire
harness.
9
Screw in the screw that connects the
Forward Fluorescence Detector to the
instrument.
Dowel pin
Cable assembly
from forward
scatter sensor
H.V. ON button
10 Press in the two latches and lift up the
top cover of the instrument.
8-4
PN 4237470AA
FORWARD FLUORESCENCE DETECTOR OPTION
INSTALL THE FORWARD FLUORESCENCE DETECTOR OPTION
11 Remove PMT4:
a.
Remove the thumbscrew in front
of PMT4.
b.
Disconnect the signal cable from
PMT4.
c.
Partially pull out PMT4.
d.
Disconnect the high voltage cable
from the bottom of PMT4.
e.
Unplug the low voltage cable from
the PMT power distribution board.
Note: PMT4 must be used because its
outer casing was modified for use with
the FFD option when your FFD option
was installed.
High voltage cable
(On bottom)
Low
voltage
cable
Signal
cable
PMT4
Low
voltage
cable
Power
distribution
board
Thumbscrew
Top view
12 Plug the corresponding three extension
cables from the FFD option kit into
PMT4:
PN 4237470AA
r
Signal cable
r
High voltage cable
r
Low voltage cable.
8-5
8
FORWARD FLUORESCENCE DETECTOR OPTION
INSTALL THE FORWARD FLUORESCENCE DETECTOR OPTION
13 Install PMT4 into the FFD:
a.
Loosen the thumbscrew on the top
of the FFD.
b.
Insert PMT4, window side down,
into the FFD.
c.
Thumbscrew
Low
voltage
cable
PMT4
Tighten the screw on the top of the
FFD.
Window
14 Plug the signal and high voltage
extension cables, from PMT4 in the
FFD, into the signal and high voltage
cables that you disconnected from
PMT4 in step 11.
Low
voltage
cable
15 Plug the extension cord for the low
voltage cable from PMT4 in the FFD
into the PMT power distribution board.
Window
Signal
cable
High
voltage
cable
Front view
16 Insert up to three filters into the FFD.
8-6
PN 4237470AA
FORWARD FLUORESCENCE DETECTOR OPTION
INSTALL THE FORWARD FLUORESCENCE DETECTOR OPTION
17 Install the two FFD sensing area
covers:
a.
Position the small cover on the
FFD and tighten its thumbscrew.
b.
Position the large cover over the
FFD.
c.
Push the large cover into the laser
cover on the right to fit them
together.
d.
Push down on the interlock rod.
e.
Tighten the thumbscrew on the
large cover.
Note: It may be easier to perform this
step with the signal cable removed.
Interlock rod
AV
DA
NG
ER
Thumbscrew
LA
ON AM
BE
IATI
RAD RE TO
R
SU
SE
PO
EX
OID
Sample
line
PN 4237470AA
Signal
Cable
8-7
8
FORWARD FLUORESCENCE DETECTOR OPTION
INSTALL THE FORWARD FLUORESCENCE DETECTOR OPTION
18 Replace the curved cover between the
sample station control panel and the
cytometer touch screen.
7470069A
19 Close the instrument:
a.
Close the top cover
b.
Replace the interlock cover.
c.
Close the front cover.
20 Power up the instrument.
21 Align the instrument if needed.
22 Look at the forward scatter mask and
ensure that it is blocking the bright
laser beam light. You should not see the
laser beam reflecting off of the forward
scatter pickup lens.
8-8
PN 4237470AA
INDEX
Numerics
300 series lasers
cooling water specifications, 3-21
dimensions and weight, 3-20
floor loading and space, 3-20
general, 3-20
laser head, description, 3-23
laser head, illustration, 3-24
power requirements, 3-21
power supply, description, 3-21
power supply, illustration, 3-22
PowerTrack feature, 3-27
remote control module, controls, 3-23
remote control module, illustration, 3-22
set laser power, 3-27
setting the aperture, 3-27
shutdown procedure, 3-28
startup procedure, 3-24
tune laser for maximum power, 3-27
70 Spectrum laser
See Spectrum laser
A
Adaptec Direc-CD
software supplied with recordable
CD-ROM, 4-2
adjust
window width for gated amp option, 6-13
air-cooled lasers
available options, 2-1
cleaning and maintenance, 2-16
dimensions and weight, 2-1
floor loading and space, 2-1
heat and ventilation, 2-1, 3-1
installation requirements, 2-1
mirror alignment, 2-1
See also argon, red HeNe, green HeNe, and
HeCd lasers
alignment
check, gated amp option, 6-11
imaging optics, 6-2
procedure for air-cooled lasers, 2-2
alignment mirror
in mirror alignment, 2-7, 3-7
amplifier speed
gated amp option, 6-11
aperture, set
300 series lasers, 3-27
PN 4237470AA
argon lasers
air-cooled, general, 2-17
air-cooled, power requirements, 2-17
air-cooled, shutdown procedure, 2-18
air-cooled, startup procedures, 2-17
assignment screen description
gated amp option, 6-6
Autoclone sorting option
calibration, 5-21
calibration after installation, 5-1
installation requirements, 5-1
microscope slide, 5-13
plate holder, 5-13
slide adapter, 5-13
sorting, 5-15
startup procedure, 5-4
B
beam path
in mirror alignment, air-cooled lasers, 2-6
in mirror alignment, water-cooled lasers,
3-6
beam reducer
in mirror alignment, 2-8, 3-9
bimorph cable
connected to standard flow cell, 7-4
connecting to large particle sorting option
flow cell, 7-5
borescope
use in imaging optics alignment, 6-4
C
calcium flux with Indo-1 and two color
markers
dual laser application, example, 6-1
calibration
after installation, Autoclone sorting
option, 5-1
Autoclone sorting option, 5-21
before you fine tune, Autoclone sorting
option, 5-21
fine tune procedure, Autoclone sorting
option, 5-22
CD-ROM
recordable, 4-1
CD-ROM, recordable
Adaptec Direc-CD software, 4-2
components included in kit, 4-1
-1
INDEX
Easy CD-Creator software, 4-2
general, 4-1
hardware recommendations, 4-1
software included, 4-2
certification label, 1-10
changing laser wavelength
water-cooled lasers, 3-1
clock
settings for gated amp option, 6-10
Coherent lasers
See Enterprise II, 300 series, or Spectrum
lasers
compression ring
how to remove, 7-14
sample tubing assembly, location, 7-14
control screen settings
gated amp option, 6-8, 6-10
controls and indicators
green HeNe lasers, 2-22
HeCd lasers, 2-26
red HeNe lasers, 2-19
conventions
used in the manual, xii
cooling water
300 series lasers, specifications, 3-21
Enterprise II series lasers, specifications,
3-15
Spectrum laser, specifications, 3-29
Cyonics argon laser
See argon lasers
D
DAPI/FITC
dual laser application, example, 6-1
data storage
backups, recommended, 4-1
delay
settings for gated amp option, 6-10
dimensions and weight
300 series lasers, 3-20
air-cooled lasers, 2-1
Enterprise II series lasers, 3-14
Spectrum laser, 3-29
water-cooled lasers, 3-14, 3-20, 3-29
documentation for your instrument
about your options manual, xi
conventions used, xii
introduction to your manuals, xi
using the manuals, xi
-2
drops delay
Autoclone sorting option, 5-8
dual laser applications
examples, 6-1
feature of the gated amp option, 6-1
E
Easy CD-Creator software
general, 4-2
installation procedures, 4-2
electrical power
300 series lasers specifications, 3-21
air-cooled argon laser specifications, 2-17
Enterprise II series lasers specifications,
3-14
green HeNe laser specifications, 2-22
HeCd laser specifications, 2-26
red HeNe laser specifications, 2-19
Spectrum laser specifications, 3-29
Enterprise II series lasers
cooling water specifications, 3-15
dimensions and weight, 3-14
floor loading and space, 3-14
general, 3-14
laser head, description, 3-17
laser head, illustration, 3-17
power requirements, 3-14
power supply, controls and indicators,
3-15
power supply, description, 3-15
power supply, illustration, 3-16
remote control module, description, 3-16
remote control module, illustration, 3-16
shutdown procedure, 3-19
startup procedure, 3-18
F
ferrule
how to remove, 7-14
sample tubing assembly, location, 7-14
FFD
See forward fluorescence detector option
filters
in mirror alignment, 2-11, 3-10
pressure, large particle sorting option,
7-10
PN 4237470AA
INDEX
fine tune
Autoclone sorting option calibration
procedure, 5-22
Autoclone sorting option calibration,
what to do before, 5-21
floor loading and space
300 series lasers, 3-20
air-cooled lasers, 2-1
Enterprise II series lasers, 3-14
Spectrum laser, 3-29
water-cooled lasers, 3-14, 3-20, 3-29
Flow-Check fluorospheres
See fluorospheres
fluorescence
compensation, combinations for gated
amp option, 6-10
fluorospheres
use for Autoclone option sort matrix, 5-8
use in imaging optics alignment, 6-5
focusing screen
illustration, 6-4
use in imaging optics alignment, 6-4
forward fluorescence detector option
description, 8-1
high voltage cable, 8-5
installation procedure, 8-1
low voltage cable, 8-5
signal cable, 8-5
use of extension cables, 8-5
use of PMT4, 8-5
forward scatter sensor
removal, 8-3
forward scatter sensor shield
removal, 8-2
full optical alignment
when to perform, 7-11
G
gate window width
adjust, gated amp option, 6-13
gated amp option
amplifier speed, 6-11
assignment screen description, 6-6
check alignment, 6-11
control screen description, 6-8
control screen settings, 6-10
description, 6-1
dual laser applications, 6-1
PN 4237470AA
fluorescence compensation combinations,
6-10
imaging optics alignment, 6-2
set up screens, 6-6
time-domain gating, 6-1
gating
time-domain, for gated amp option, 6-1
green HeNe lasers
controls and indicators, 2-22
general, 2-22
laser head, illustration, 2-23
power requirements, 2-22
power supply, illustration, 2-23
shutdown procedure, 2-25
startup procedure, 2-24
H
hazards
biological, description of, 1-1
biological, list of, 1-1
heat and ventilation
air-cooled lasers, 2-1
water-cooled lasers, 3-1
HeCd lasers
applications, 2-26
components, 2-26
controls and indicators, 2-26
general, 2-26
power requirements, 2-26
shutdown procedure, 2-30
startup procedure, 2-29
helium cadium laser
See HeCd lasers
helium neon laser
See red HeNe or green HeNe lasers
high voltage cable
forward fluorescence detector option, 8-5
PMT4, 8-5
Hoechst/Chromomycin
dual laser application, example, 6-2
HPSS
where to find information about this
option, xi
HyPerSort System
See HPSS
-3
INDEX
I
imaging optics
alignment procedure, 6-2
Innova lasers
See Enterprise II, 300 series, or Spectrum
lasers
installation
Autoclone sorting option, calibration
procedure, 5-1
requirements, air-cooled lasers, 2-1
requirements, Autoclone option, 5-1
requirements, water-cooled lasers, 3-1
Iomega jaz drive
See jaz drive
Iomega Zip drive
See Zip drive
J
jaz drive
general, 4-2
L
label
CDRH-approved, 1-2
certification, 1-10
laser classification, outer cover, 1-9
laser warning, Cyonics argon, 1-3
laser warning, detector and sensing
compartment, 1-9
laser warning, Enterprise II series, 1-5
laser warning, FFD sensing area, 1-8
laser warning, green HeNe, 1-4
laser warning, HeCd, 1-5
laser warning, Innova 300 series, 1-6
laser warning, Innova 70 Spectrum, 1-6
laser warning, list of, 1-2
laser warning, rear cover, 1-10
laser warning, red HeNe, 1-4
laser warning, sensing area, 1-7
large particle sorting option
components, 7-1
install the flow cell, 7-1
pressure filter assembly, 7-10
sample tubing assembly, 7-6
laser
certification label, 1-10
changing wavelength, 3-1
classification label, outer cover, 1-9
-4
dual applications, gated amp option, 6-1
set power for 300 series lasers, 3-27
set power for Spectrum laser, 3-34
tune for maximum power for 300 series
lasers, 3-27
tune for maximum power for Spectrum
laser, 3-35
warning label, Cyonics argon laser, 1-3
warning label, detector and sensing
compartment, 1-9
warning label, Enterprise II series, 1-5
warning label, FFD sensing area, 1-8
warning label, green HeNe, 1-4
warning label, HeCd, 1-5
warning label, Innova 300 series, 1-6
warning label, Innova 70 Spectrum, 1-6
warning label, rear cover, 1-10
warning label, red HeNe, 1-4
warning label, sensing area, 1-7
laser arrangement
air-cooled lasers, example, 2-2
example, water-cooled lasers, 3-2
laser head
300 series lasers, description, 3-23
300 series lasers, illustration, 3-24
Enterprise II series lasers, description,
3-17
Enterprise II series lasers, illustration,
3-17
green HeNe lasers, illustration, 2-23
HeCd lasers, illustration, 2-28
red HeNe lasers, illustration, 2-19
Spectrum laser, illustration, 3-32
low voltage cable
forward fluorescence detector option, 8-5
PMT4, 8-5
lower beam lockout
feature of the gated amp option, 6-1
settings for gated amp option, 6-10
LPS
See large particle sorting option
M
manuals
additional, xi
from manufacturer of an option, xi, 2-26,
3-13, 3-14, 3-20, 3-29
manuals for your instrument
about your options manual, xi
PN 4237470AA
INDEX
conventions used, xii
how to use, xi
introduction to your manuals, xi
using the manuals, xi
Melles Griot laser
See green HeNe or red HeNe lasers
microscope slide
use with Autoclone sorting option, 5-13
mirror alignment
air-cooled lasers, description, 2-1
air-cooled lasers, procedure, 2-2
water-cooled lasers, description, 3-2
water-cooled lasers, procedure, 3-2
O
Omnichrome laser
See HeCd lasers
options manual
conventions used, xii
introduction to your manuals, xi
using the manuals, xi
P
pinhole carrier
illustration, 6-5
use in imaging optics alignment, 6-5
plate characteristics
for Autoclone sorting option, 5-17
plate holder
use with Autoclone sorting option, 5-13
PMT4
high voltage cable, 8-5
install into the forward fluorescence
detector option, 8-6
low voltage cable, 8-5
removal, 8-5
signal cable, 8-5
used with forward fluorescence detector
option, 8-5
PMTs
power distribution board, 8-5
power distribution board
for PMTs, location, 8-5
power requirements
300 series lasers, 3-21
air-cooled argon lasers, 2-17
Enterprise II series lasers, 3-14
green HeNe lasers, 2-22
PN 4237470AA
HeCd lasers, 2-26
red HeNe lasers, 2-19
Spectrum laser, 3-29
power supply
300 series lasers, description, 3-21
300 series lasers, illustration, 3-22
Enterprise II series lasers, controls and
indicators, 3-15
Enterprise II series lasers, description,
3-15
Enterprise II series lasers, illustration,
3-16
green HeNe lasers, illustration, 2-22
HeCd lasers, control settings, 2-27
HeCd lasers, description, 2-26
HeCd lasers, illustration, 2-27
HeCd lasers, indicators, 2-28
Spectrum laser, controls and indicators,
3-30
Spectrum laser, description, 3-29
Spectrum laser, illustration, 3-30
PowerTrack feature
300 series lasers, 3-27
pressure filter assembly
install for large particle sorting option,
7-10
location, 7-10
R
red HeNe lasers
controls and indicators, 2-19
general, 2-19
illustration, laser head, 2-19
power requirements, 2-19
shutdown procedure, 2-21
startup procedure, 2-20
remote control module
300 series lasers, controls, 3-23
300 series lasers, description, 3-22
300 series lasers, illustration, 3-22
Enterprise II series lasers, controls, 3-17
Enterprise II series lasers, description,
3-16
Enterprise II series lasers, illustration,
3-16
Spectrum laser, controls and indicators,
3-30
Spectrum laser, description, 3-31
Spectrum laser, illustration, 3-31
-5
INDEX
replace
pressure filter assembly, 7-10
sample tubing assembly, large particle
sorting option, 7-12
S
safety precautions
biohazard safety, 7-3
biological hazards, 1-1
electrical, 7-12
laser safety, 1-1
radiation hazards, 1-2
sample
run with Autoclone sorting option, 5-15
sample tubing assembly
compression ring, 7-14
ferrule, 7-14
large particle sorting option, 7-6, 7-12
tubing connector to sample pickup cap,
7-14
when to replace, 7-12
sensing area
forward fluorescence detector option
cover, 8-7
gaining access to, 7-2, 7-13, 8-1
location, 7-2, 7-13, 8-1
sheath air filter
connected to pressure filter assembly, 7-10
shutdown
procedure for 300 series lasers, 3-28
procedure for air-cooled argon lasers,
2-18
procedure for Enterprise II series lasers,
3-19
procedure for green HeNe lasers, 2-25
procedure for HeCd lasers, 2-30
procedure for red HeNe lasers, 2-21
procedure for Spectrum laser, 3-35
signal cable
forward fluorescence detector option, 8-5
PMT4, 8-5
signal delay
settings for gated amp option, 6-10
silicone tubing
large particle sorting option, 7-7
simultaneous four-color clone study
dual laser application, example, 6-2
slide adapter
use with Autoclone sorting option, 5-13
-6
sort
matrix, Autoclone option, 5-8
settings, for Autoclone sorting option,
5-18
sorting
Autoclone sorting option, 5-15
Spectrum laser
cooling water specifications, 3-29
dimensions and weight, 3-29
floor loading and space, 3-29
general, 3-28
laser head, illustration, 3-32
power requirements, 3-29
power supply, controls and indicators,
3-30
power supply, description, 3-29
power supply, illustration, 3-30
remote control module, controls and
indicators, 3-30
remote control module, description, 3-31
remote control module, illustration, 3-31
set laser power, 3-34
shutdown procedure, 3-35
startup procedure, 3-33
tune laser for maximum power, 3-35
startup
procedure for 300 series lasers, 3-24
procedure for air-cooled argon lasers,
2-17
procedure for Autoclone sorting option,
5-4
procedure for Enterprise II series lasers,
3-18
procedure for green HeNe lasers, 2-24
procedure for HeCd lasers, 2-29
procedure for red HeNe lasers, 2-20
procedure for Spectrum laser, 3-33
storage media
magnetic, care of, 4-1
options, 4-1
storage options
descriptions, 4-1
T
time-domain gating
feature of the gated amp option, 6-1
trigger discriminator
settings for gated amp option, 6-10
PN 4237470AA
INDEX
trigger gain
settings for gated amp option, 6-10
tubing connector
sheath and vacuum, large particle sorting
option, 7-6
sheath and vacuum, standard flow cell,
7-5
to sample pickup cap, location, 7-14
to standard flow cell body, 7-4
U
Uniphase laser
See red HeNe laser
W
water, cooling
See cooling water
water-cooled lasers
changing laser wavelength, 3-1
cleaning and maintenance, 3-13
dimensions and weight, 3-29
floor loading and space, 3-14, 3-20, 3-29
general, 3-1
heat and ventilation, 3-1
installation requirements, 3-1
mirror alignment, 3-2
operation, 3-1
See also Enterprise II, 300 series, and
Spectrum lasers
well timeout
for Autoclone sorting option, 5-17
width
settings for gated amp option, 6-10
Y
Yamaha CD-ROM
See CD-ROM, recordable
Z
Zip drive
components included in kit, 4-1
description, 4-1
general, 4-1
PN 4237470AA
-7
INDEX
-8
PN 4237470AA
TRADEMARKS
The BECKMAN COULTER logo, AccuSort, ALTRA, ALTRA HyPerSort, Autoclone,
COULTER, Elite, EPICS, EXPO, FlowCentre, FlowCheck, SYSTEM II, and XL are trademarks
of Beckman Coulter, Inc.
All other trademarks, service marks, products, or services are trademarks or registered
trademarks of their respective holders.
PN 4237470AA
COULTER EPICS ALTRA Flow Cytometry Documentation
s
Reference
PN 4237469
Use and Function • Installation • Operation Principles •
Specifications/Characteristics • Precautions/Hazards • Optical Filters • Glossary •
Index
s
Operator’s Guide
PN 4237468
Controls and Indicators • Software Overview • Startup • Quality Control • Run
Samples • Data Analysis • Sorting • Shutdown • Define Protocol • Data
Management and DOS • Prism, Ratio and Time • Metafiles • Color Adjusts •
Panelyzer • Time of Flight • Pulse Pile Up • Index
s
Special Procedures and
Troubleshooting
PN 4237467
Calibration • Cleaning Procedures • Replacement/Adjustment Procedures •
Troubleshooting • Index
s
Options Manual
PN 4237470
Safety Precautions • Air-Cooled Lasers • Water-Cooled Lasers • Storage Options •
Autoclone Sorting Option • Gated Amp Option • Large Particle Sorting Option •
Forward Fluorescence Detector Option • Index
s
FlowCentre Multimedia
Workstation - Getting Started
PN 4237415
Overview • Starting ALTRA or Elite Software • Starting XL/XL-MCL SYSTEM II
Software from the Startup Menu • Starting XL/XL-MCL SYSTEM II Software from
an Icon • System Components • Maintaining System Performance • Software
Support
Come visit us at www.beckmancoulter.com
Printed on Recycled Paper
Copyright © Beckman Coulter, Inc. 2000, 2010
All Rights Reserved
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