Roper Photometric User manual

-
User Manual for Evolve 512
© Copyright 2009
Photometrics
3440 East Britannia Drive
Tucson, Arizona 85706
Tel: 800.874.9789/520.889.9933
Fax: 520.295.0299
All rights reserved. No part of this publication may be reproduced by any means without the written permission of
Photometrics, a division of Roper Scientific, Inc.
Printed in the United States of America.
Acrobat and Reader are registered trademarks of Adobe Systems Incorporated in the United States and/or other countries.
Pentium is a registered trademark of Intel Corporation.
PVCAM, Saguaro, and Photometrics are registered trademarks of Roper Scientific, Inc.
Mac, Macintosh and FireWire are trademarks of Apple Computer, Inc., registered in the U.S. and other countries.
Windows and Windows XP are registered trademarks of Microsoft Corporation in the United States and/or other
countries.
Other brand and product names are the trademarks or registered trademarks of their respective owners and manufacturers.
The information in this publication is believed to be accurate as of the publication release date. However, Photometrics
does not assume any responsibility for any consequences including any damages resulting from the use thereof. The
information contained herein is subject to change without notice. Revision of this publication may be issued to
incorporate such change.
Evolve 512 Users Manual
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Customer
Service
If you have any questions about your camera system, please contact Photometrics
Customer Service. When you call, please have your Photometrics part number or
equipment serial numbers available.
USA
Photometrics
3440 East Britannia Drive
Tucson, Arizona 85706
tel: 800.874.9789 or
520.889.9933
fax: 520.295.0299
email: cservice@photomet.com
GERMANY
Roper Scientific, GmbH
Rosenheimer Landstr. 87
D-85521 Ottobrunn, Germany
tel: 49.89.660.779.3
fax: 49.89.660.779.50
email: support@roperscientific.de
ii
BENELUX
Roper Scientific, BV
Ir. D.S. Tuijnmanweg 10
4131 PN VIANEN, Netherlands
tel: 31.347.324989
fax: 31.347.324979
email: mailto@roperscientific.com
JAPAN
Nipon Roper, K.K.
D-10E 1-3 Nakase,
Mihama-ku, Chiba-shi
Japan 261-8501
tel: 81.43.274.8022
fax: 81.43.274.8023
email: sales@roper.co.jp
FRANCE
Roper Scientific, SARL
Z.I. Petite Montagne Sud
4, rue de l'Oisans - C.E. 1702
91017 Evry Cedex, France
tel: 33.160.86.03.65
fax: 33.160.86.07.09
email: princeton.instruments@wanadoo.fr
UK
Photometrics UK Ltd.
Beech House
27 Little Marlow Road
Marlow
Buckinghamshire
SL7 1HA
Tel: 44 1628 890858
Fax: 44 1628 898381
email: UKsupport@photomet.co.uk
Evolve 512 Users Manual
LIMITED WARRANTY
Photometrics, a division of Roper Scientific, Inc., (“Photometrics,” us,” “we,” “our”) makes the following limited
warranties. These limited warranties extend to the original purchaser (“You”, “you”) only and no other purchaser or
transferee. We have complete control over all warranties and may alter or terminate any or all warranties at any time we
deem necessary.
Basic Limited One (1) Year Warranty
Photometrics warrants this product against substantial defects in materials and / or workmanship for a period of up to one
(1) year after shipment. During this period, Photometrics will repair the product or, at its sole option, repair or replace any
defective part without charge to you. You must deliver the entire product to the Photometrics factory or, at our option, to
a factory-authorized service center. You are responsible for the shipping costs to return the product. International
customers should contact their local Photometrics authorized representative/distributor for repair information and
assistance, or visit our technical support page at www.photomet.com.
Limited One (1) Year Warranty on Refurbished or Discontinued Products
Photometrics warrants, with the exception of the CCD imaging device (which carries NO WARRANTIES EXPRESS OR
IMPLIED), this product against defects in materials or workmanship for a period of up to one (1) year after shipment.
During this period, Photometrics will repair or replace, at its sole option, any defective parts, without charge to you. You
must deliver the entire product to the Photometrics factory or, at our option, a factory-authorized service center. You are
responsible for the shipping costs to return the product to Photometrics. International customers should contact their local
Photometrics representative/distributor for repair information and assistance or visit our technical support page at
www.photomet.com.
Normal Wear Item Disclaimer
Photometrics does not warrant certain items against defect due to normal wear and tear. These items include internal and
external shutters, cables, and connectors. These items carry no warranty, expressed or implied.
Sealed Chamber Integrity Limited 24 Month Warranty
Photometrics warrants the sealed chamber integrity of all our products for a period of twenty-four (24) months after
shipment. If, at anytime within twenty-four (24) months from the date of delivery, the detector should experience a sealed
chamber failure, all parts and labor needed to restore the chamber seal will be covered by us. Open chamber products
carry NO WARRANTY TO THE CCD IMAGING DEVICE, EXPRESSED OR IMPLIED. Responsibility for shipping
charges is as described above under our Basic Limited One (1) Year Warranty.
Vacuum Integrity Limited 24 Month Warranty
Photometrics warrants the vacuum integrity of all our products for a period of up to twenty-four (24) months from the
date of shipment. We warrant that the detector head will maintain the factory-set operating temperature without the
requirement for customer pumping. Should the detector experience a Vacuum Integrity failure at anytime within twentyfour (24) months from the date of delivery all parts and labor needed to restore the vacuum integrity will be covered by
us. Responsibility for shipping charges is as described above under our Basic Limited One (1) Year Warranty.
Software Limited Warranty
Photometrics warrants all of our manufactured software discs to be free from substantial defects in materials and / or
workmanship under normal use for a period of one (1) year from shipment. Photometrics does not warrant that the
function of the software will meet your requirements or that operation will be uninterrupted or error free. You assume
responsibility for selecting the software to achieve your intended results and for the use and results obtained from the
software. In addition, during the one (1) year limited warranty. The original purchaser is entitled to receive free version
upgrades. Version upgrades supplied free of charge will be in the form of a download from the Internet. Those customers
who do not have access to the Internet may obtain the version upgrades on a CD-ROM from our factory for an incidental
shipping and handling charge. See Item 12 in the following section of this warranty ("Your Responsibility") for more
information.
Owner's Manual and Troubleshooting
You should read the owner’s manual thoroughly before operating this product. In the unlikely event that you should
encounter difficulty operating this product, the owner’s manual should be consulted before contacting the Photometrics
technical support staff or authorized service representative for assistance. If you have consulted the owner's manual and
the problem still persists, please contact the Photometrics technical support staff or our authorized service representative.
See Item 12 in the following section of this warranty ("Your Responsibility") for more information.
Evolve 512 Users Manual
iii
Your Responsibility
The above Limited Warranties are subject to the following terms and conditions:
1. You must retain your bill of sale (invoice) and present it upon request for service and repairs or provide other
proof of purchase satisfactory to Photometrics.
2.
You must notify the Photometrics factory service center within (30) days after you have taken delivery of a
product or part that you believe to be defective. With the exception of customers who claim a “technical issue”
with the operation of the product or part, all invoices must be paid in full in accordance with the terms of sale.
Failure to pay invoices when due may result in the interruption and/or cancellation of your one (1) year limited
warranty and/or any other warranty, expressed or implied.
3.
All warranty service must be made by the Photometrics factory or, at our option, an authorized service center.
4.
Before products or parts can be returned for service you must contact the Photometrics factory and receive a
return authorization number (RMA). Products or parts returned for service without a return authorization
evidenced by an RMA will be sent back freight collect.
5.
These warranties are effective only if purchased from the Photometrics factory or one of our authorized
manufacturer's representatives or distributors.
6.
Unless specified in the original purchase agreement, Photometrics is not responsible for installation, setup, or
disassembly at the customer’s location.
7.
Warranties extend only to defects in materials or workmanship as limited above and do not extend to any
product or part which has:
been lost or discarded by you;
been damaged as a result of misuse, improper installation, faulty or inadequate maintenance or failure
to follow instructions furnished by us;
had serial numbers removed, altered, defaced, or rendered illegible;
been subjected to improper or unauthorized repair; or
been damaged due to fire, flood, radiation, or other “acts of God” or other contingencies beyond the
control of Photometrics.
8.
After the warranty period has expired, you may contact the Photometrics factory or a Photometricsauthorized representative for repair information and/or extended warranty plans.
9.
Physically damaged units or units that have been modified are not acceptable for repair in or out of
warranty and will be returned as received.
10. All warranties implied by state law or non-U.S. laws, including the implied warranties of merchantability
and fitness for a particular purpose, are expressly limited to the duration of the limited warranties set forth
above. With the exception of any warranties implied by state law or non-U.S. laws, as hereby limited, the
forgoing warranty is exclusive and in lieu of all other warranties, guarantees, agreements, and similar
obligations of manufacturer or seller with respect to the repair or replacement of any parts. In no event
shall Photometrics’ liability exceed the cost of the repair or replacement of the defective product or part.
11. This limited warranty gives you specific legal rights and you may also have other rights that may vary
from state to state and from country to country. Some states and countries do not allow limitations on
how long an implied warranty lasts, when an action may be brought, or the exclusion or limitation of
incidental or consequential damages, so the above provisions may not apply to you.
12. When contacting us for technical support or service assistance, please refer to the Photometrics factory
of purchase, contact your authorized Photometrics representative or reseller, or visit our technical
support page at www.photomet.com.
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Evolve 512 Users Manual
Evolve 512 Users Manual
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Evolve 512 Users Manual
Table of Contents
Chapter 1 Overview Introduction ...............................................................................................................................1 Technology Introduction .........................................................................................................1 System Components ................................................................................................................2 About This Manual ..................................................................................................................3 Precautions.................................................................................................................................3 Environmental Requirements .................................................................................................4 Storage Requirements...............................................................................................................4 Microscopes, Lenses, and Tripods .........................................................................................4 Repairs ........................................................................................................................................4 Cleaning .....................................................................................................................................4 Chapter 2 System Installation Introduction ...............................................................................................................................5 Software Compatibility Requirements ...................................................................................5 Host Computer Requirements ................................................................................................6 Multiple Cameras......................................................................................................................6 Software Installation .................................................................................................................6 Installing the FireWire Interface Card..................................................................................7 Connecting Your Evolve 512 Camera ....................................................................................7 To connect your Evolve 512 camera ....................................................................................7 Chapter 3 Operating Features Introduction ...............................................................................................................................9 On-Chip Multiplication Gain ................................................................................................9 Offset (bias) ..........................................................................................................................9 Exposure- Readout Modes ....................................................................................................9 Non-Overlap Mode .............................................................................................................10 Example: Non-Overlap Mode ............................................................................................11 Overlap Mode (Simultaneous Exposure-Readout) ............................................................12 Triggered Operation............................................................................................................14 Trigger-first Mode ..............................................................................................................14 Strobe Mode .......................................................................................................................15 Bulb Mode ..........................................................................................................................16 Conversion Gain .................................................................................................................16 Readout Speed (Typical) ....................................................................................................17 Binning ...............................................................................................................................17 Evolve 512 Features ................................................................................................................18 Vari-Bit ...............................................................................................................................18 Quant-View ........................................................................................................................19 Rapid-Cal ............................................................................................................................20 Background Event Reduction Technology .........................................................................21 Black-Lock / Top-Lock ......................................................................................................22 Dual-Readout Port Operation .............................................................................................23 Conversion Gain ................................................................................................................25 Readout Speed (typical) ......................................................................................................25 Binning ...............................................................................................................................25 Evolve 512 Application Examples......................................................................................26 Evolve 512 Users Manual
vii
Application Settings Summary ...........................................................................................26 Chapter 4 Troubleshooting System Does Not Boot Normally .......................................................................................27 New Hardware Found Dialog Box Does Not Appear (Windows 2000/XP) .......................27 Images Not Displayed........................................................................................................28 Camera Running Too Warm ...............................................................................................28 PVCAM Error Message Appears........................................................................................28 Lengthy Pauses During Imaging.........................................................................................28 Evolve 512 Will Not Image When Attached to Certain Dual-processor or Hyperthreadingcapable PCs.........................................................................................................................28 Chapter 5 Basic Specifications Evolve 512 Front/Side Views ...............................................................................................29 Camera Weight ...................................................................................................................30 CCD Specifications .................................................................................................................30 Connectors ...............................................................................................................................30 Power Connector Pinout: ....................................................................................................31 Power Supply Specifications ................................................................................................33 Appendix
Liquid Cooled Evolve Setup Instructions ...........................................................................................34
Index................................................................................................................................. 35 Figures
Figure 1. Comparison of Traditional CCD and Evolve CCD Array Structures ..............2 Figure 2. FireWire interface Card ...........................................................................................2 Figure 3. DATA Cable ..............................................................................................................7 Figure 4. Evolve 512 Side Panel .............................................................................................8 Figure 5. Non-overlap Mode Operational Sequence.........................................................10 Figure 6. Required Settings for Non-Overlap Mode Operation ......................................11 Figure 7. Timing Diagram for Non-Overlap Mode ...........................................................11 Figure 8. Required Settings for Overlap Mode Operation ...............................................12 Figure 9. Timing Diagram for Overlap Mode when Exposure Time < Readout Time 13 Figure 10. Timing Diagram for Overlap Mode when Exposure Time > Readout Time14 Figure 11. Trigger-First Mode Timing Diagram: Overlap Mode.....................................14 Figure 12. Trigger-First Mode Timing Diagram: Non-Overlap Mode ...........................15 Figure 13. Strobe Mode Timing Diagram ...........................................................................15 Figure 14. Bulb Mode Timing Diagram ..............................................................................16 Figure 15. Benefit of Varibit ...................................................................................................18 Figure 16. DAC Setting vs. Charge Multiplication Gain for Evolve 512 ........................20 Figure 17. RapidCal Status.....................................................................................................21 Figure 18. BERT Off vs BERT On ..........................................................................................22 Figure 19. Evolve 512 CCD Array Structure .....................................................................23 Figure 20. Software Selection of Readout Port ...................................................................24 Figure 21. Evolve 512 Front View .......................................................................................29 Figure 22. Evolve 512 Side View .........................................................................................29 Figure 23. Evolve 512 Side Panel..........................................................................................30 Figure 24. Power Connector Pinout.....................................................................................31 Figure 25. Input/Output Connector....................................................................................32 Figure 26. Power Supply Front ............................................................................................33 Figure 27. Liquid Cooled Evolve Setup Diagram ...............................................................34 viii
Evolve 512 Users Manual
Tables
Table 1.
Table 2.
Table 3.
Table 4.
Evolve 512 Users Manual
Evolve 512 Readout Speeds and Typical Conversion Gains .............................25 Application Type and Evolve 512 Setup Parameters .........................................26 CCD Specifications ..................................................................................................30 Input/Output Definitions ......................................................................................32 ix
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Evolve 512 Users Manual
Chapter 1
Overview
Introduction
The Photometrics® Evolve camera employs an advanced CCD technology called "impact
ionization" to achieve charge multiplication gain above the read noise. As a result, the
cameras offer unparalleled sensitivity even at high frame rates.
Evolve cameras are most suited for high speed and/or low-light level imaging applications
such as single molecule fluorescence, ion imaging, and adaptive optics. With the traditional
readout amplifier, the Evolve can also be used for standard fluorescence and imaging
applications requiring higher dynamic range.
Currently the Evolve line consists of the following camera model:

Evolve 512
o
Uses a 512x512 Back-illuminated CCD with 16x16 µm pixels.
o
Has two readout amplifiers – one that offers charge multiplication gain
and another for traditional readout.
Technology
Introduction
The principal difference between a charge-multiplying CCD and a traditional CCD is the
presence of an extended serial register in the new device (see Figure 1). Electrons are
accelerated from pixel to pixel in the extended portion of the serial register (also referred to
as a multiplication register) by applying higher-than-typical CCD clock voltages. This
causes secondary electrons to be generated in the silicon by impact ionization. The degree of
multiplication gain is controlled by increasing or decreasing clock voltages for this register
(gain is exponentially proportional to the voltage). Although the probability of generating
secondary electrons is fairly low (typically 0.01 per stage), over the large number of stages of
a typical multiplication register, the total gain can be quite high.
This technology combines the ease of use and robustness of a traditional CCD with the gain
capabilities of an intensified CCD in a single device. The combination of this technology
with frame-transfer readout makes the Evolve cameras excellent choices for experiments
where fast framing and low light sensitivity are required.
Evolve 512 Users Manual
1
Figure 1. Comparison of Traditional CCD and Evolve CCD Array Structures
The Evolve 512 also offers a non-multiplication readout similar to the one available in
traditional CCD cameras. This makes these more versatile cameras that can be used for
standard imaging applications requiring higher dynamic range.
System
Components
The Evolve 512 package includes PVCAM® drivers and Micro-Manager imaging software
along with a Firewire cable, a Firewire Interface PCI card, and a test lens.
Figure 2. FireWire interface Card
2
Evolve 512 Users Manual
About This
Manual
The Evolve 512 User Manual is divided into five chapters. It is suggested that you read the
entire manual before operating the camera in order to ensure proper use. The chapter
contents are briefly described below.
Note: The information in these chapters applies only to the Evolve 512 and is currently not applicable
to any other Photometric camera.

Overview — This is a basic overview of the Evolve 512 that includes precautions
and general maintenance/equipment information.

System Installation — Instructions for connecting your Evolve 512 camera to
your computer via the Evolve FireWire interface card.

Operating Features — Discusses Evolve 512 features such as on-chip
multiplication gain, readout, and trigger modes. Provides additional information
on dual-readout mode feature available.

Troubleshooting — Provides answers to camera system problems.

Basic Specifications — Provides specifications for Evolve 512 system
components.
Precautions
The CCD and other system electronics are extremely sensitive to electrostatic discharge
(ESD). To avoid permanently damaging the system, please observe the following
precautions:
Evolve 512 Users Manual

If you are using high-voltage equipment (such as an arc lamp) with your camera
system, be sure to turn the camera power on last and power the camera off first.

Never connect or disconnect any cable while the system is powered on.

Although you should switch off the camera’s power before disconnecting any
camera system cable, you do not need to power off your computer to detach the
cables.

Use caution when triggering high-current switching devices (such as an arc
lamp) near your system. The CCD can be permanently damaged by transient
voltage spikes. If electrically noisy devices are present, an isolated, conditioned
power line or dedicated isolation transformer is highly recommended.

Always leave one inch of space around the camera’s external cooling fins for
airflow.

Never open the camera. There are no user-serviceable parts inside the Evolve 512
camera. Opening the camera voids the warranty.

Use only the FireWire interface card, cables, and power supply designated for
this camera system. Using non-Evolve cables, FireWire interface cards, or power
supplies may result in permanent damage to your system.

Do not use a C-mount lens with optics that extend behind the lens flange.
3
Environmental
Requirements
The Evolve 512 camera system should be operated in a clean, dry environment. The camera
system’s ambient operating temperature is 0°C to 30°C with 80% relative humidity, noncondensing.
Storage
Requirements
Store the Evolve 512 camera system in its original containers. To protect the system from
excessive heat, cold, and moisture, store at an ambient temperature between -20°C and 60°C
with a relative humidity of 0% to 90%, noncondensing.
Microscopes,
Lenses, and
Tripods
The camera has a standard threaded video mount and can be mounted to any microscope
that accepts a standard C-mount adapter. The camera also allows you to install any lens that
is compatible with a standard threaded video mount as long as its optics do not extend
behind the flange of the lens. The Evolve 512 camera can be mounted to a tripod using the
tripod mounting attachment located on the sides of the camera.
The Evolve 512 C-mount adapter is designed so it can be screwed in or out to change the
focal depth. Once the focal depth as been changed, the adapter is secured by two setscrews.
See Adjusting the C Mount Adapter (Chapter 2) for more information.
Repairs
The Evolve 512 camera system contains no user-serviceable parts. Repairs must be done by
Photometrics. Should your camera system need repair, contact Photometrics Customer
Service. Please save the original packing materials so you can safely ship the camera system
to another location or return it for repairs if necessary.
Note: Do not open the camera. Opening the Evolve 512 camera voids the warranty.
Cleaning
Clean exterior surfaces of the camera with a dry, lint-free cloth. To remove stains, contact
Photometrics Customer Service. To clean the camera’s imaging window, use only a filtered
compressed-air source. Hand-held cans are not recommended, as they may spray propellant
onto the window. Do not touch the window.
4
Evolve 512 Users Manual
Chapter 2
System Installation
Carefully review the Precautions section in the previous chapter before performing any of the
procedures outlined here. Again, use only an Evolve data cable and an Evolve FireWire interface card
with your Evolve 512 camera. Using a different cable or interface card may result in permanent
damage to your system.
Introduction
Your Evolve 512 camera system has the following hardware components:

Camera head

FireWire interface card

Data cable

Power supply with power cord

Red test lens

CD-ROM

Quick installation Guide
Evolve 512 system components are linked by the data cable and controlled by your host
computer system. All of these hardware components should be included with your
shipment. Refer to the information and figures in the System Components section (Chapter
1).
Keep all the original packing materials so you can safely ship the Evolve 512 system to
another location or return it for service if necessary.
If you have any difficulty with any step of the instructions, call Photometrics Customer
Service.
Software
Compatibility
Requirements
The Evolve 512 package includes the PVCAM drivers and the Micro-Manager software
program designed for use with this camera.
All other imaging software must also be PVCAM-compatible. For full access to Evolve 512
hardware functions, the current version of PVCAM must be used.
Evolve 512 Users Manual
5
Host Computer
Requirements
The host computer (PC) for your Evolve 512 camera must have the following:

Windows® XP (SP2) operating system

1 GHz Pentium® 4 (or greater)

256 MB RAM (or greater)

CD-ROM drive

At least one unused traditional PCI or PCI-X card slot

16-bit color display (or greater)
Note: The above requirements are the minimum for operating an Evolve camera. A faster computer
with larger memory (RAM) will greatly enhance the software performance during live mode
operations.
If you are a Mac® user, the host computer for your Evolve 512 camera must have the
following:

Macintosh® OS X operating system (Power PC based)

G4/G5 computer

256 MB RAM (or greater)

CD-ROM drive

At least one unused traditional PCI or PCI-X card slot

Video adapter that supports 24-bit color (millions of colors)
Multiple
Cameras
PVCAM supports multiple open cameras. In order to use this function, it must also be
supported by your imaging software.
If your imaging software supports multiple cameras, there must be a separate interface card
for each camera.
Software
Installation
An Installation Guide appropriate to your system is included as an insert with the camera
system. This guide provides step-by-step instructions for installing the camera interface
software and the application software for Windows-based computers. Additional
instructions are included for installing a FireWire interface card in your computer and
capturing images.
The Photometrics CD-ROM contains the following files.
6

MacOS directory — contains required files for installing on a Macintosh
computer.

Manuals directory — contains user manuals in PDF format.

WinOS directory — contains the files for installing on a Windows PC.

Linux directory — (not applicable to Evolve 512).
Evolve 512 Users Manual
Installing the
FireWire
Interface Card
You will be using an Evolve FireWire interface card to allow the camera to communicate
with your computer.
Before installing the interface card, refer to the ‘Release Notes’ files on the CD-ROM and to
the Software Installation insert. The insert instructions will step you through the FireWire
interface card installation.
After installing the interface card, continue to Connecting Your Evolve 512 Camera.
Connecting Your
Evolve 512
Camera
The following connectors are located on the back of the Evolve 512 camera:

DATA connector: Type 1, 6-pin IEEE-1394a (FireWire) connector for data transfer

POWER connector: 25-pin, Dsub connector for camera power (see POWER
Connector Pinout section in Chapter 5 for details)

Power switch: Turns the camera on and off.
The following connector is located on the power supply of the Evolve 512 camera:

I/O connector: Hirose HR10A-10R-10S, 10-pin connector for input/output
control signals (see I/O Connector Pinout later in this manual)
To connect your
Evolve 512
camera
1.
Connect the straight end of the DATA cable to the Evolve FireWire interface card
that you have installed in the host computer. The DATA cable (shown below)
connects your Evolve 512 camera to the Evolve FireWire interface card. This
interface cable is identifiable by its 6-pin connectors. It is designed to serve as a
conduit for data.
Figure 3. DATA Cable
Evolve 512 Users Manual
7
2.
Connect the right-angled end of the DATA cable to the FireWire port (indicated
with a red arrow in next graphic) located on the side of the camera.
Figure 4. Evolve 512 Side Panel
8
3.
Verify that the power switch on the side of the camera is in the off position.
4.
Connect the power supply to the Power connector on the side of the camera.
5.
Plug the power cord into the power supply and then into a suitable wall outlet.
6.
Switch on the camera (power switch on the side of camera).
Evolve 512 Users Manual
Chapter 3
Operating Features
Introduction
The features described in this section are identical in all Evolve 512 cameras unless
otherwise stated.
On-Chip
Multiplication
Gain
As explained previously, the Evolve 512 uses a unique CCD capable of multiplying the
charge (electrons) generated in the pixels. When the multiplication is sufficiently high, it is
possible to see extremely low-light events. The amount of multiplication is controlled by the
voltage applied to multiplication register clocks. These are certified by a quantitative
multiplication gain slider which has 0-1000 steps. The calibration feature of the camera
maps the gain slider to a voltage which will provide the electron multiplication level
selected.
Offset (bias)
CCD cameras are typically designed to produce a certain level of offset (also known as bias)
when no light is present and the exposure time is set to zero (0). Typically, the user
subtracts an offset (bias) from the sample image for quantitative measurement. Since the
offset can change based on several factors such as multiplication gain, speed, etc., it is
recommended that a fresh offset (bias) image be taken with the same settings as the sample
image and then be subtracted from the sample image.
The Evolve 512 has an automatically adjusting offset switch. When a speed/gain setting is
altered, the bias valve is kept as close to a pre-selected offset as possible. This enhances the
quantitative stability of the camera.
ExposureReadout Modes
The Evolve 512 camera uses a frame transfer CCD and supports Non-Overlap and Overlap
exposure and readout modes. These modes are further described in the sections that follow.
Note: A frame transfer CCD has both a light-sensitive area (sensor area) and a storage area (frame
transfer area). Since the image can be quickly transferred from the sensor area to the frame transfer
area, there is no need for a mechanical shutter.
Evolve 512 Users Manual
9
Non-Overlap
Mode
Non-Overlap mode allows you to expose the array for the exposure time specified in the
software and is similar in performance to a normal, full-frame device. The operational
sequence for this mode is:
1. Clearing the CCD,
2.
Exposing for the specified exposure time,
3.
Shifting the image from the sensor area to the frame-transfer area, and
4.
Reading out the CCD.
Steps 1-4 are repeated for each frame in a sequence. Steps 1 and 3, clearing the CCD and
shifting the image, are usually very short and typically have minimum impact on the frame
rate.
Figure 5. Non-overlap Mode Operational Sequence
In PVCAM compatible software, this sequence is programmed by simply setting:

clocking mode to “Frame transfer” and

clearing mode to “Pre-exposure” with clear count of “1”(or more).
Figure. 6 shows these settings on a software dialogue box.
Note: Since the software you are using may show the settings differently, you should refer to the
software documentation for accurate information.
10
Evolve 512 Users Manual
Select these parameters to
operate your Evolve 512
camera in Non-Overlap
mode.
Figure 6. Required Settings for Non-Overlap Mode Operation
Example:
Non-Overlap
Mode
Operation in Non-Overlap mode is illustrated in the timing diagram below. In this example,
the exposure time is 10 ms and the readout time is 29.5 ms. The total time to take 3 frames is
118.5 ms (3  10 ms + 3 29.5 ms), equivalent to a frame rate of 25.3 fps (3 frames  0.1185
seconds).
Note: The exposure and readout times listed are for illustration purpose only. Actual values may vary.
Refer to the product data sheet for the actual readout times.
Figure 7. Timing Diagram for Non-Overlap Mode
Referring to figure 7, it can be seen that exposure and readout are carried out in nonoverlapped fashion. As a result, each frame in the sequence is precisely exposed for the time
specified (i.e. 10 ms).
Evolve 512 Users Manual
11
Overlap Mode
(Simultaneous
Exposure-Readout)
Overlap mode is extremely useful in applications requiring continuous imaging (100% duty
cycle). Once a frame is exposed and transferred into the frame transfer area, the next
exposure immediately starts and continues until the previous frame is read out or until the
exposure time is finished, whichever is longer (so the minimum effective exposure time in
this mode is the readout time). This mode of operation allows you to continuously image a
specimen to obtain better kinetic information about a process. Figure 8 shows the required
Evolve 512 settings for overlap mode. Note that the clearing mode is set to "Pre-sequence"
(as opposed to "Pre-exposure" in Non-Overlap mode).
Figure 8. Required Settings for Overlap Mode Operation
Note: In Overlap mode, the minimum effective exposure time is the readout time.
The simultaneous exposure-readout mechanism is illustrated with two examples.
Example 1: Overlap Mode when Exposure Time < Readout Time
Consider a situation where full frame readout is 29.5 ms, the exposure time is 10 ms, and
three frames are taken in overlap mode. The first frame is exposed precisely for the length of
time entered into the software (10 ms) and all subsequent frames are exposed for the readout
time. The total time to acquire 3 frames is then 98.5 ms (3  29.5 ms + 10 ms), equivalent to a
frame rate of 30.45 fps (3 frames  0.0985 seconds).
Note: Because the first frame is exposed for 10 ms and the others for 29.5 ms, the first frame may look
less bright compared to all other frames.
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Evolve 512 Users Manual
In Overlap mode when exposure time < readout time, the total time (TN) taken to capture N
frames is given by:
TN = (tR  N) + texp
Where
TN = Total time taken to capture a sequence of N frames
tR = readout time for one frame
N = total number of frames in a sequence
texp = exposure time
Figure 9. Timing Diagram for Overlap Mode when Exposure Time < Readout Time
Example 2: Overlap Mode when Exposure Time > Readout Time
If the exposure time is set to 50 ms with the readout time remaining at 30 ms, the time taken
to acquire 3 frames will be 179.5 ms (3  50 ms + 29.5 ms), which is equivalent to a frame rate
of 16.7 fps.
In Overlap mode when exposure time > readout time, the total time (TN) taken to capture N
frames is expressed as:
TN = (texp  N) + tR
Where
TN = Total time taken to capture a sequence of N frames
texp = exposure time
N = total number of frames in a sequence
tR = readout time for one frame
From the timing diagram, you can see that because the exposure time is greater than the
readout time, all frames are precisely exposed for the duration entered into the software and
have similar intensities.
Evolve 512 Users Manual
13
Figure 10. Timing Diagram for Overlap Mode when Exposure Time > Readout Time
Triggered Operation
The Evolve 512 camera offers several methods of integration with external trigger sources,
such as delay generators or laser pre-triggers. Each camera has a 10-pin, Hirose HR10A I/O
connector (pinout functions are described in Chapter 5) on the power supply for trigger
input/out and various TTL input and output operations. A special cable is available from
Photometrics to access primary signals such as "Trigger input," "Expose out," "Frame
readout," and "Shutter out." In the default mode, the camera triggers on the rising edge of a
TTL signal. Evolve 512 cameras support the trigger modes described in the next sections.
Trigger-first Mode
In this mode, the camera requires only one trigger to acquire a sequence of frames. The
camera can operate in overlap or non-overlap mode depending on the settings explained in
the previous section. Once the initial trigger is received, the camera ignores any further
triggers until the entire exposure/readout sequence is completed (see Figures 11 and 12,
which show a three-frame sequence).
Ignored
Trigger
Clears
#1
#2
#3
#1
#2
Exposure
#3
Readout
Figure 11. Trigger-First Mode Timing Diagram: Overlap Mode
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Evolve 512 Users Manual
Ignored
Trigger
Clears
#1
#2
#3
Exposure
#1
#2
#3
Readout
Figure 12. Trigger-First Mode Timing Diagram: Non-Overlap Mode
Strobe Mode
In Strobe mode, each frame in a sequence requires a trigger. Each frame is exposed for the
length of time entered into the software and is then read out. If a trigger arrives during the
exposure-readout of the previous frame, it is ignored (see Figure 13). For a sequence of one
frame, strobe mode and trigger-first mode are the same. The Trigger Ready signal goes high
when Read Out is completed. It goes low when the Trigger signal is accepted and exposure
is started. The shaded areas
denote the idle time between exposures. Strobe mode only
operates in Non-Overlap mode.
Figure 13. Strobe Mode Timing Diagram
Evolve 512 Users Manual
15
Bulb Mode
In Bulb mode, exposure time for each frame is determined by the trigger pulse width.
Exposure time entered into the software is ignored in this mode (see Figure 14). If a trigger
arrives during the readout of the previous frame, it is ignored. The Trigger Ready signal
goes high when Read-out has been completed. It then goes low when the Trigger signal is
accepted and exposure is started. The shaded areas
denote the idle time between
exposures. Bulb mode only operates in Non-Overlap mode.
Figure 14. Bulb Mode Timing Diagram
Notes:
1. When the clocking mode is "Alternate Frame Transfer", the camera can be operated in "Overlap" or
"Non-Overlap" mode as explained previously.
2. Trigger-first mode allows overlap or non-overlap modes. In strobe and bulb triggering modes, the
camera can be operated in non-overlap mode only.
Conversion Gain
Apart from multiplication gain, which actually multiplies the number of electrons generated
in each pixel, the Evolve 512 also has three (3) conversion gains. Conversion gain simply
quantifies how the resulting electrons are converted into ADU (Analog-to-Digital units) and
displayed on the display screen.
Typical conversion gains for the readout port are listed later in this chapter. Your camera is
individually calibrated for these gains. So, please refer to the certificate of performance that
accompanied your camera.
Note: As the on-chip multiplication introduces additional noise, it is recommended that the
multiplication be used only as required. For more information, refer to the "On-Chip Multiplication
Gain" technical note. This technical note can be accessed by going to the Photometrics web site at
www.photomet.com, clicking on "Library", clicking on "Technical Notes", and selecting the title from
the list of notes.
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Evolve 512 Users Manual
Readout Speed
(Typical)
Evolve 512 cameras are implemented with 10 MHz, 5MHz and 1.25 MHz digitization speeds
(10 million, 5 million and 1.25 million pixels per second, respectively). At 10 MHz it gives
approximately 31 frames per second (fps), whereas 1.25 MHz operation offers better image
quality. With either digitizer, increased frame rate can be achieved by choosing a sub-region.
For more information, please refer to the product data sheets.
The camera uses a frame-transfer CCD with a built-in storage (frame transfer) area, which
makes simultaneous exposure and readout possible (Overlap mode) and which allows
precise exposures to be set (Non-Overlap mode). These two readout modes are further
explained in the "Exposure - Readout Modes" section in this chapter.
Binning
Binning (combining pixels into one super pixel) allows you to increase the sensitivity and
frame rate. On the other hand, binning reduces spatial resolution. The Evolve 512 allows
binning of 1, 2, 4, and 8 in the serial (horizontal) direction and 1 to 256 binning in the parallel
direction.
Evolve 512 Users Manual
17
Evolve 512 Features
Photometrics has developed several new industry-leading camera capabilities to streamline
user workflow and enable researchers to concentrate directly on the image data that are
important to their studies. These advanced functions enhance the quantitative nature of the
camera while simultaneously allowing researchers to home in on the data they are looking
for.
IMPORTANT NOTE: All of these features can be turned off in order to return to a
standard-type setup for anyone who would rather use the camera in a more traditional
way.





Advanced Evolve™ features include:
Quant-View™
Rapid-Cal™
Background Event Reduction Technology™
Black-Lock™ / Top-Lock™
Vari-Bit™
This document contains a description of each of these features -- what they actually do and
how they can be utilized by the research community to make their data much more
quantitative.
Vari-Bit
This feature allows camera users to decide at which bit depth the digitization of the image
should occur. Often with EMCCD imaging in biology, the intrascene dynamic range does
not justify the use of high-bit-depth digitization (e.g., 16 bits) and the resultant image
contains gray levels that represent fractions of photoelectrons. With this advanced feature,
users can determine and select a more relevant digitization bit depth for their experiments.
How will it help?
By matching the digitization bit depth more closely to the actual intrascene dynamic range,
the image quality will improve. Actual transitions between light levels from pixel to pixel
will appear sharper to the eye; subsequently, users will be more certain of the results they
are seeing. Using higher-bit-depth digitization does not make measurements less
quantitative when measuring a scene with a low dynamic range; however, it does introduce
many extra (or unnecessary) gray levels that do not represent actual physical photons.
Figure 15. Benefit of Varibit
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Evolve 512 Users Manual
Quant-View
A CCD counts photons by generating electrons when incident photons hit the device’s
array. These electrons are counted and converted into an analog-to-digital unit (ADU) value.
High-performance camera companies such as Photometrics provide users the system gain
number (expressed as electrons per ADU) for each of their cameras, enabling them to
convert the ADU value back into actual electrons.
The new Quant-View feature performs this conversion for users in real-time. The camera
reads out the pixel values in electrons measured. This number, along with the exposure
time, can then be used to calculate the number of electrons generated at any pixel per unit
measure of time – effectively providing an electron flux measurement.
Electrons-per-gray-level selector
This allows users to select how many electrons will cause a single gray-level increase in the
image data. If there are enough photons to saturate the 16-bit data bus but there is still no
saturation of the physical pixel of the EMCCD, then users can raise the number of electrons
needed to increase the gray level to the next value.
For example, if you have 1000 electrons, then you are able to decide if this should be
represented by 1000 gray levels (1 electron per gray level selected), by 500 gray levels (2
electrons per gray level selected), or by 250 gray levels (4 electrons per gray level selected).
This enables users to utilize Quant-View while maintaining the entire dynamic range of the
EMCCD. The camera’s highest bit depth in this quantitative mode is 16 bits. This means that
as many as 65,535 gray levels per pixel can be sent over the bus. However, for the CCD97
used in the Evolve 512 camera, the total number of electrons possible in a single pixel can
easily reach 200,000. Thus, if the camera was set at 1 electron per gray level, then the pixel
would come across as saturated (it would max out at the 65,535 value). So, by selecting each
gray level to be equivalent to 4 electrons, the camera will send over a value of 50,000 gray
levels, which is equivalent to 200,000 electrons. This feature is part of the Quant-View
feature, ensuring that users are able to utilize the full capability of the EMCCD while
remaining in Quant-View mode.
How will Quant-View help?
By actually measuring the electrons generated, users quantitatively know what the image is
measuring in actual electrons. There are many examples where such data will be important
for bio-research. One example would be when normalizing for transfection levels. Often,
fluorescent proteins are transfected at different levels and results are taken and measured.
These results are often just reported relative to “control cells”. Utilizing Quant-View
measurements will enable researchers to discern if they are looking at cells that are
transfected to similar levels, day to day and even year to year. Over-expression artifacts and
results could be identified in this manner, leading to higher-quality research.
Quant-View in action
For further explanation, including a real-time demonstration of Quant-View, please visit
www.evolve-emccd.com
Evolve 512 Users Manual
19
Rapid-Cal
Calibration
The Evolve 512 has incorporated a calibration routine which allows the camera to adjust
voltages of the ADC offsets such that the electron multiplication gain input given to the
camera reflects the actual gain provided by the device. Settings 1 to 1000 on the electron
multiplication gain slider are mapped linearly to provide the actual multiplication gain
requested by the user. Due to the impact-ionization method used with the electronmultiplication gain register the actual gain realized by the detector with time will slowly be
reduced. By using the calibration routine the camera is able to re-establish the electron
multiplication gain slider such that it will provide the gain which is input on the gain slider.
This ensures the quantitative nature of the camera over time. Even though the camera is
capable of delivering large multiplication gain factors, multiplication gain should only be
used as needed to preserve as much dynamic range as necessary and to prolong the device’s
lifetime.
Figure 16. DAC Setting vs. Charge Multiplication Gain for Evolve 512
EMCCD cameras are subject to aging of the EMCCD register as a result of its usage. The
Evolve 512 has a simple calibration feature that performs the industry’s most accurate EM
calibration within 3 minutes. A simple turn of the cameras nose-piece closes a shutter and
activates a light source which the detector uses to calibrate its EM gain. This ensures that
users will receive the most accurate electron multiplication and the EM gain applied
matches what the user requests. Simple software control will allow the user to use this
feature as a manual shutter in order to block all light from the sensor in order to take dark
reference frames if necessary.
How will Rapid-Cal help?
It is a necessary requirement that your electron multiplication be accurate if you wish to be
quantify the images taken by an EMCCD camera. The evolve cameras Rapid-Cal allows
extremely accurate and rapid calibration of the EM-Gain ensuring your data can be
quantitative all of the time. It is recommended that calibration be performed once a week
although the requirements of calibration will vary depending on the usage of the camera.
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Evolve 512 Users Manual
Rapid-Cal in action
Figure 17. RapidCal Status
For further explanation, including a real-time demonstration of Rapid-Cal, please visit
www.evolve-emccd.com
Background Event Reduction Technology
EMCCD cameras are actually capable of detecting single photons. However, the real
detection limit of these cameras is set by the number of background events. These can arise
from two things, Dark-current (which is thermal generation of an electron and is a
temperature-dependent phenomenon) and also clock induced charge (CIC) electrons (also
called spurious charge). Each can lead to the generation of non-photon derived electrons
which are multiplied through the electron-multiplication register generating random high
value pixels which are above the read noise. These are often referred to as “speckles” in the
image. Photometrics has developed this real-time feature to help users identify such events
and correct for them, in their images in real-time.
Users can set a threshold parameter that will measure the variation of a pixel value from all
of its neighboring pixels and if the pixel value is unusually large such that it cannot of come
from real light through the microscope then the evolve camera will realign the pixel value to
a best approximation of where it should be – all in real-time. This serves to absolutely
minimize the influence of clock induced charge and thermally induced amplified events on
acquired data.
How will background event reduction help?
This technology will help reduce the EMCCD induced artifact of spurious noise from
entering your acquired data. Some may see this as a processing feature and claim that data
is being altered. It is important to note that this feature (like all others) is completely
controlled by the user and can be switched on or off. It should also be noted that the
camera’s major task is to most accurately represent the sample it is imaging. If the
technology components used in the camera can cause the introduction of artifacts in the
image then utilizing technology to minimize the influence of such artifacts is also just an
extension of good technology design. The BERT function is an example of additional
technology being used to enable the camera to more accurately represent the data being
acquired.
Evolve 512 Users Manual
21
1000X EM Gain (no BERT)
1000X EM Gain (BERT=1.2)
Figure 18. BERT Off vs BERT On
Background event reduction in action
For further explanation, including a demonstration of background event reduction
technology please visit www.evolve-emccd.com
Black-Lock / Top-Lock
Black-Lock
This advanced feature effectively functions as an intensity filter for the camera. It allows
users to select the background level at which the EMCCD will make all pixels at or below
the selected value equal to the value selected.
How will it help?
This advanced feature allows users to define an effective floor for an image at a level which
they select. In the case of the Evolve 512, many users operate the camera in EM gain mode
and apply EM gain at high levels. In such scenarios, EMCCD cameras can end up having
multiple ADUs per electron. Traditional CCDs are set up to have electrons per ADU, which
makes sense, as you can take the ADU measurement and convert it back to electrons.
However, in the case of an EMCCD when EM gain is applied, the actual gain can increase
such that you will end up with multiple ADUs being equivalent to a single electron.
For example, a traditional EMCCD camera with 3 electrons per ADU and 300x EM gain
applied actually has 0.01 electrons per ADU. Thus, 100 ADUs is equivalent to a single
electron of signal. For most bio-researchers, any signal less than 1 electron (equivalent to a
single photon in a 100% QE system) does not represent meaningful data.
With the Black-Lock feature from Photometrics, users can select at which point this value
should be set. For instance, in the above example, one could set the Black Lock at 75 ADU
above the image bias. As a result, the camera will read all pixels at or below 75 ADU as 75
ADU. Effectively, users are able to set the floor of the image and determine at which
intensity levels data will be collected.
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Evolve 512 Users Manual
Top-Lock
This advanced feature effectively functions as an intensity filter for the camera. It allows
users to select the background level at which the EMCCD will make all pixels at or above
the selected value equal to the value selected.
How will it help?
This advanced feature allows users to define an effective roof for an image at a level which
they select. In many experimental conditions, there may be intense objects in the field of
view that are actually of little or no interest to the experimenter. A simple example may be a
piece of debris in the field of view that happens to be highly fluorescently labeled. If the
debris is not of interest, a top intensity value of the image can be selected so that every
single pixel in the image at or above the selected top intensity value will be displayed as the
selected top value.
Another example where Top-Lock may be important is when researchers are looking for
small, low-intensity-labeled features that are actually budding off or being released from a
large feature. A specific example would be something like small vesicles being released from
the Golgi apparatus in a cell. If the researcher is only interested in observing and tracking
these features but the highly labeled Golgi is swamping the image display, then the camera
can be set to lock all intensities above a certain level to that level. This will make the features
the researcher is interested in much more visible, as well as scaled nicely within the display
image for ready visualization.
Combining Black-Lock and Top-Lock
Utilizing Top-Lock and Black-Lock together allows users to narrow visualization rapidly
and effectively into the intensity range of the image features they are most interested in.
Black-lock and Top-lock in action
For further explanation, including a real-time demonstration of black-lock and top-lock,
please visit www.evolve-emccd.com
Dual-Readout
Port Operation
The Evolve 512 is configured with software-selectable dual-readout amplifiers, also referred
to as "ports", shown in the next figure. The two amplifiers are listed below.
Port #1: On-chip Multiplication amplifier
Port #2: Traditional or non-multiplication amplifier
Figure 19. Evolve 512 CCD Array Structure
Evolve 512 Users Manual
23
The software allows port selection. An example of a software dialogue to select the ports is
shown in the next figure.
Readout Port
Selection.
Figure 20. Software Selection of Readout Port
Note: As the user interface may be different, please refer to your software manual for specific
information on readout amplifier/port selection.
On-Chip Multiplication Amplifier (Port #1): Operation of multiplication gain in the Evolve
512 is different from the previous generation of these cameras. The new “Evolve” generation
of cameras have new functionality that was not available on previous models. A setting of 0
(zero) results in unity gain and the subsequent settings are linearly related to the actual
multiplication gain. Since the multiplication gain can be used to overcome the read noise of
the fast amplifier, this mode is most useful in applications requiring low-light sensitivity at
high frame rates (e.g., Single molecule fluorescence, ion imaging, etc.).
Note: When used with a standard lens, Port #1 (multiplication port) produces an image in the correct
orientation. The first pixel is read out at the bottom left of the array.
Traditional or Non-Multiplication amplifier (Port #2): When the camera is using this
amplifier, electrons (signal) generated in pixels are clocked through the standard serial
register. The amplifier is designed to take advantage of the dynamic range of the CCD and is
most useful when the frame rate is not critical, (e.g., bright field, fixed cell fluorescence, etc.).
Note: Since the first pixel to be read out from Port #2 is at the bottom right (closest to the Port #2), the
resulting image is a mirror image of the same image if it were read out of Port #1. Some software
packages do a "horizontal-flip" in the software when Port #2 is selected to ensure the correct
orientation for all ports. Please check your software documentation.
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Evolve 512 Users Manual
Conversion
Gain
Apart from multiplication gain, which actually multiplies the number of electrons generated
in each pixel, the Evolve 512 also has three (3) conversion gains. Conversion gain simply
quantifies how the resulting electrons are converted into ADU (Analog-to-Digital units) and
displayed on the display screen.
Typical conversion gains for different ports are listed in Table 1. Your camera is individually
calibrated for these gains. So, please refer to the certificate of performance that accompanied
your camera.
Readout Speed
(typical)
The Evolve 512 has two readout speeds available on each of the two ports as listed in the
table below:
Readout Amplifier (Port)
Port #1 (Multiplication)
Readout Speeds
5 MHz
10 MHz
Port #2 (Non-multiplication)
1.25 MHz
5 MHz
Conversion Gain
(e- /ADU)
#1: 12e- /ADU
#2: 6e- /ADU
#3: 3e- /ADU
#1: 4e- /ADU
#2: 2e- /ADU
#3: 1e- /ADU
Table 1. Evolve 512 Readout Speeds and Typical Conversion Gains
A slower readout speed can be used when better noise performance is needed at the expense
of frame rate. On the other hand, increased frame readout rate can be achieved by one or
more of the following:

higher readout speed,

sub-region selection,

binning.
For more information on frame rate, please refer to the product data sheet.
Binning
Binning (combining pixels into one super pixel) allows you to increase the sensitivity and
frame rate. On the other hand, binning reduces spatial resolution. The Evolve 512 allows
binning of 1, 2, 4 and 8 in the serial (horizontal) direction and binning in the parallel
(vertical) direction, up to 256.
Note: Due to the small array size in the Evolve 512 edge artifacts may be present at higher binning.
Evolve 512 Users Manual
25
Evolve 512
Application
Examples
Example 1:
“I would like to obtain kinetic information (time-correlated) information of the single molecule
fluorescence. My light level is fairly low and I want to optimize the frame rate of the camera.”
For this application, the "Multiplication" port (Port #1) should be selected and camera speed
should be set to 10 MHz. In addition, the camera should be operated in “Overlap mode”.
This can be achieved by setting the camera to “frame transfer mode” with clearing mode
“Pre-sequence” and number of clears “1” (refer to Figure 8). If possible, choose a sub-region
(ROI) and/or binning to further increase the frame rate. Finally, on-chip multiplication gain
can be used to boost the signal level and achieve a high signal-to-noise ratio.
Example 2:
“My application requires precise control of the exposure time (less than the readout time) and I want
to operate the camera in the most sensitive mode possible.”
For better imaging characteristics, the "Multiplication" port (Port #1) is selected and camera
speed must be set to 5 MHz. Use on-chip multiplication gain for increased sensitivity. To
achieve an exposure time less than readout time, use Non-Overlap mode (shown in Figure
6) and set the clearing mode to “Pre-exposure” and clear count to “2”.
Example 3:
“I would like to image a bright target/sample (e.g., fluorescence or bright field) that is fixed. High
resolution and high dynamic range are important."
For imaging bright, non-moving samples, the "Non-multiplication" port (Port #2) offers the
best performance in terms of dynamic range. Set the speed to 1.25 MHz (or 5 MHz, if higher
speed is required) and conversion gain to #2 (this allows maximum full well of the CCD to
be used). Reading out the full CCD (no-ROI or binning) gives best resolution possible. In
addition, camera can be operated in overlap or non-overlap mode as explained before.
Application Settings
Summary
The table below summarizes typical Evolve 512 settings for two types of applications and is provided
as a setup aid. Actual settings will vary based on the exact nature of your experiment design.
Application
Readout
Amplifier
(Port)
Readout
Speed
Multiplication
Gain
Conversion
Gain
(e-/ADU)
Readout Mode
Kinetic samples
(single molecule
fluorescence or
tracking)
Multiplication 10 MHz or Use as required to #2: 6e- /ADU
(Port #1)
5 MHz
obtain best signalto-noise ratio
(SNR). Typical
EM gain values
are around 200.
Overlap mode
(Frame Transfer,
Pre-Sequence)
Fixed samples
(bright field)
Non1.25 MHz
multiplication
(Port #2)
#1: 4e- /ADU
Overlap or Nonoverlap
Not Applicable
Table 2. Application Type and Evolve 512 Setup Parameters
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Evolve 512 Users Manual
Chapter 4
Troubleshooting
If you have any difficulty while troubleshooting, or do not see your camera system’s symptoms listed
here, contact Photometrics Customer Service.
System Does Not
Boot Normally
If your operating system does not boot normally after you have installed an interface card,
try installing the new card in another open PCI slot. If this does not work:
1. Turn off your computer and remove the newly installed interface card.
2.
Turn your computer back on. If your system boots normally, there is probably an
interrupt conflict between a previously installed expansion card and the interface
card that you are installing.
3.
If you need assistance resolving the interrupt conflict, contact Photometrics
Customer Service.
New Hardware Found Dialog
Box Does Not Appear
(Windows 2000/XP)
If the New Hardware Found dialog box does not appear after installing a new interface card
to your computer and booting Windows 2000/XP:
Evolve 512 Users Manual

Check to make sure that the new interface card is inserted in an expansion slot
according to your computer manufacturer’s instructions and that the Evolve 512
system’s CD-ROM disc is in the host computer’s CD drive.

It is possible that there is a conflict between the new interface card and a
previously installed expansion card. With the computer’s power turned off,
remove any previously installed expansion cards that your system does not need
to function. (If you are unsure which cards can be safely removed, call
Photometrics Customer Service.) Then turn your computer back on and boot
Windows 2000/XP again.

If the New Hardware Found dialog box still does not appear, contact
Photometrics Customer Service.
27
Images Not
Displayed
If no images appear:

Confirm that the camera switch is set to on.

Confirm that the Evolve 512 camera is selected in your imaging software
application.

Power off the camera and the host computer and check all system connections
(particularly the DATA and power cables). Restart.

Confirm that operating system is set for at least 64k colors (16 bits).

Confirm that the camera is operational by taking an image with a standard
C-mount lens attached to your Evolve 512. Using normal room lighting, place the
camera on a table about 3 meters away from an object and acquire an image.
If the problem persists, contact Photometrics Customer Service.
Camera Running
Too Warm
It is normal for the camera to be slightly warm to the touch while in operation. However, if
the camera is more than slightly warm to the touch (and at least one inch of space has been
left around the external cooling fins for airflow), switch off the camera immediately and
contact Photometrics Customer Service.
PVCAM Error
Message Appears
If a PVCAM error message appears, note the message’s number code and contact
Photometrics Customer Service.
Lengthy Pauses
During Imaging
If you notice lengthy pauses marked by a lot of disk activity while imaging:

Close any other programs that may be running.

Install more physical memory (RAM) in your computer system.
Evolve 512 Will Not
Image When Attached
to Certain Dual-processor
or Hyperthreading-capable PCs
Some high-end Windows 2000/XP-enabled computers with Intel Pentium or Xeon™
processors include a memory enhancement called Physical Address Extension (PAE). PAE
allows these computers to address memory above 4GB; however, when PAE is enabled the
IEEE-1394 Evolve 512 camera will not transmit any pixel data.
To determine whether your computer is running with PAE enabled, go to the Control Panel
and open the "System" icon. Under the "General" tab, you will find a reference to Physical
Address Extension in the "Computer" section.
To disable PAE, you must edit your C:\boot.ini file and remove the /PAE flag.
28
Evolve 512 Users Manual
Chapter 5
Basic Specifications
Evolve 512
Front/Side Views
Figure 21. Evolve 512 Front View
Figure 22. Evolve 512 Side View
Evolve 512 Users Manual
29
Camera Weight
Weight: 6.5 lbs. (2.8 kg)
CCD Specifications
Window
UV grade fused-silica
Broadband MgF2 anti-reflective
coating on both surfaces
CCD Array
CCD
e2v CCD97-00
CCD Process
Back Illuminated
Resolution
512 x 512
Pixel Size
16 µm x 16 µm
Readout Amplifiers (Ports)
2
Digitalization (Readout)
Rate
10 MHz, 5 MHz, 1.25 MHz
Table 3. CCD Specifications
Connectors
Figure 23. Evolve 512 Side Panel
The following connectors are located on the side of the camera.


30
connector: Type 1, 6 pin IEEE-1394a (FireWire).
POWER connector: 25-pin, Dsub connector for camera power and I/O
functions.
Evolve 512 Users Manual
Power Connector
Pinout:
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.
RX-IO
SHUTTER OUT
FPGA+5V
SPARE IO-1
/TRIG
/A PWR CONTROL
TEC-PWR
TEC-PWR
-15V
+37V
+3.3V
+3.3V
+16V
FPGA-GND
SPARE IO-2
EXPOSE
TX-IO
EOF
GND
TEC GND
TEC CONTROL
TEC GND
+ 37V
GND
GND
Evolve 512 Users Manual
Figure 24. Power Connector Pinout
31
I/O Connector
Pinout
The I/O (Input/Output Status) connector located on the power supply provides TTL level
trigger and status functions. Inputs must be at least 3.15 V for a high and less than 0.9 V for a
low.
The numbers on the I/O connector diagram correspond to the numbers given to the
definition of each of the pins. The I/O connector is a female, Hirose HR10A connector. An
I/O cable (Part # 37-513-001) to access Trigger Input (Pin 1), Frame Readout (Pin 3), Camera
exposing output (Pin 2), and Shutter Output (Pin 4) is available.
Figure 25. Input/Output Connector
INPUT/OUTPUT (I/O) TABLE
#
I/O INPUT
DESCRIPTION
1
Trigger Input
This input is internally tied high through a 4.7k resistor. A
rising edge of the Trigger Input signal initiates the trigger. The
trigger source would normally hold this input low and then drive
it high to initiate the trigger.
2
Camera Exposing
Output
Active high. A high level on this output indicates that the
camera is exposing (integrating).
3
Frame Readout
Active high. A high level on this output indicates that data is
being transferred.
4
Shutter Output
TTL output for timing of external shutter driver. Signal is high
during Shutter Open Delay and exposure time. The pin does
not provide power to drive the shutter directly, so an external
shutter drive controller is required.
5
Spare Input
Reserved
6
Trigger Ready
This output notifies accessory devices that the camera is ready
to receive a trigger.
7
TX Out
Accessory Serial Transmit
8
RX IN
Accessory Serial Receive
9
Power Status
A high level on this output indicates that the camera power is
switched on (+5 V = on, 0 V = off).
10
GND
System digital ground. Any external circuitry intended to
interface with the trigger control signals must reference this
ground connection.
Table 4. Input/Output Definitions
32
Evolve 512 Users Manual
Power Supply
Specifications
Voltage Input:
100-240 V~ @ 50-60 Hz
Current Input:
2.0 A
Voltage Output:
+3.6 V @ 1 A
+17 V @ 0.5 A
+37 V @ 0.2 A
-15 V @ 0.3 A
6-15 V @ 3.8 A
Maximum Power Output: 150 W
Power Supply Weight: 5 lb (2.267 kg)
Supply Cable Length: 6 ft. / 1.828 m
Note: CE certification applies to the Evolve 512 only when the camera system is operated with a CEapproved power supply
Figure 26. Power Supply Front
Evolve 512 Users Manual
33
Appendix
Liquid Cooled Evolve:
Setup Instructions
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
Unpack circulating chiller and insulated tubing assembly.
Confirm chiller and tubing are pre-filled with yellow-colored coolant. (Chiller is pre-set for 0C liquid temperature;
consult chiller user manual to change setting.)
Align both disconnects on tubing into female disconnects on chiller. NOTE: The tubing assembly has two different
sized boots on the ends. The end with the smaller diameter boot connects to the circulating chiller.
Press one connector into its mate on the chiller and twist 1/8 turn to lock.
Repeat with the second connector.
Slide the boot down over the collar surrounding the connectors on the chiller as far as it will go.
Secure around collar with zip-tie provided and repeat for other side of boot around hose insulation.
Unpack camera head, power supply, PCI card and interface cable.
Inspect components and set-up according to quick-start guide for Evolve.
DO NOT power on camera.
Slide large boot back along tubing to access connectors.
Press each on to its mating connector on the camera; listen for the “click”.
Pull each connector to ensure they are locked.
Return the boot to its original position and insert into the back of the camera as far as it will go.
Inspect the set-up to insure hoses are not pinched and boots are secure at chiller and camera.
Plug-in chiller and turn on.
Open cap on top of chiller when running to observe liquid level and confirm circulation. (Liquid surface will appear
agitated with normal circulation.)
Turn on camera power and continue setup per quick-start guide. NOTE: DO NOT turn-on camera power without
liquid circulating!
Open imaging software to set target temperature (-100C.)
Allow approximately 30 minutes to stabilize at target temperature.
Figure 27. Liquid Cooled Evolve Setup Diagram
34
Evolve 512 Users Manual
Index
B O Bias (Offset), 9
Binning, 17, 25
Bulb mode, 16
On-chip multiplication gain, 9
Overlap readout mode, 12
C Cleaning
exterior camera surfaces, 4
imaging window, 4
C-mount, 8
adjusting, 8
Computer (host), 6
Conversion gain, 24
Customer Service, ii
D Data connector, 7, 30
Digitization speeds, 17
F P Power connector, 7, 8, 30
Power Supply, 3, 5, 8, 33
Power switch, 7
Precautions, 3
PVCAM, 5, 6
R Readout modes
non-overlap, 10
overlap, 12
Readout speed, 17
Repairs, 4
S I/O connector, 7
Saguaro software, 2, 5
Software, iii, 5
Specifications, 29
Strobe mode, 15
System components, 2
L T Lenses, 4
Liquid Cooled, 34
Trigger modes
bulb, 16
strobe, 15
trigger-first, 14
Trigger-first mode, 14
Tripods, 4
FireWire, 3, 5, 7
I M Microscopes, 4
Multiplication
gain, 1
port, 16
register, 1
N W Warranty, iii
Your responsibility, iv
Non-overlap readout mode, 10
Evolve 512 Users Manual
35
57-519-001 Rev A3