Andor Technology Dragonfly 500 Hardware Manual

Dragonfly 500

Version 1.5 revised 20 December 2018

andor.com

Hardware Guide

© Andor Technology Ltd. 2018.

Dragonfly 500

TABLE OF CONTENTS

SECTION 1: INTRODUCTION ............................................................................................................. 16

1.1 TECHNICAL SUPPORT .......................................................................................................... 16

1.2 DISCLAIMER .......................................................................................................................... 17

1.3 COPYRIGHT AND PROTECTIVE NOTICES ........................................................................... 17

1.4 TRADEMARKS AND PATENT INFORMATION ....................................................................... 17

1.5 SUPPLIED COMPONENTS .................................................................................................... 18

1.6 OPTIONAL COMPONENTS .................................................................................................... 19

1.7 RECOMMENDED EMISSION FILTERS .................................................................................. 19

SECTION 2: PRODUCT OVERVIEW ................................................................................................... 20

2.1 AN INTRODUCTION TO THE DRAGONFLY .......................................................................... 20

2.1.1 System Integration .................................................................................................... 20

2.1.2 Outline of the Functionality of Imaging Modes* ...................................................... 21

2.2 SYSTEM CONFIGURATIONS ................................................................................................. 23

2.3 MICROSCOPE REQUIREMENTS .......................................................................................... 23

2.4 CAMERA ORIENTATION REQUIREMENTS ........................................................................... 23

2.5 EXTERNAL FEATURES OF THE DRAGONFLY 500 ............................................................... 24

2.5.1 Connections Panel and Status Display .................................................................... 25

2.5.2 Power Supply ............................................................................................................ 26

2.5.3 Laser Safety Interlock System ................................................................................. 27

2.5.3.1 Interlock Summary Diagram .................................................................... 28

SECTION 3: INSTALLATION ............................................................................................................... 29

3.1 LOCATION AND MOUNTING ................................................................................................. 29

3.2 VENTILATION ......................................................................................................................... 29

3.3 ASSEMBLY ............................................................................................................................. 29

3.4 TRANSPORT AND STORAGE ................................................................................................ 30

2 Version 1.5 rev 20 Dec 2018

Dragonfly 500

3.5 LASER CLASSIFICATION OF AN INSTALLED SYSTEM ....................................................... 30

SECTION 4: OPERATION ................................................................................................................... 32

4.1 EMERGENCY MAINS DISCONNECTION .............................................................................. 32

4.2 POWER-UP SEQUENCE ........................................................................................................ 32

4.3 POWER-DOWN SEQUENCE .................................................................................................. 33

4.4 RISK MITIGATION .................................................................................................................. 33

4.4.1 Mechanical Housings ............................................................................................... 33

4.4.2 Hazards Due to Moisture or Liquids ........................................................................ 33

4.5 USING THE DRAGONFLY 500 ............................................................................................... 34

4.5.1 Stage Cover............................................................................................................... 34

4.5.2 Controlling the Dragonfly 500 through the Software ............................................... 34

4.6 ADDITIONAL PROCEDURES ................................................................................................. 35

4.6.1 Removal of the Emission Filter Wheel ..................................................................... 35

4.6.2 Insertion of the Emission Filter Wheel ..................................................................... 38

4.6.3 Replacement of Emission Filters in the Filter Wheel ............................................... 39

4.6.4 Exchanging Emission Splitting Dichroics .............................................................. 40

4.6.5 Camera XY Adjustment ............................................................................................ 42

SECTION 5: MAINTENANCE .............................................................................................................. 46

5.1 CLEANING AND DECONTAMINATION .................................................................................. 46

5.2 REGULAR CHECKS ............................................................................................................... 46

5.3 ANNUAL ELECTRICAL SAFETY CHECKS ............................................................................ 46

5.4 EXTENDED WARRANTY ........................................................................................................ 46

SECTION 6: TROUBLESHOOTING .................................................................................................... 47

6.1 TROUBLESHOOTING EXAMPLES ........................................................................................ 47

6.1.1 Problem Reporting Form .......................................................................................... 52

SECTION 7: TECHNICAL SPECIFICATIONS ..................................................................................... 53

7.1 GENERAL SPECIFICATIONS ................................................................................................. 53

7.2 ENVIRONMENTAL SPECIFICATIONS .................................................................................... 53

7.3 POWER SUPPLY REQUIREMENTS ....................................................................................... 53

Version 1.5 rev 20 Dec 2018 3

Dragonfly 500

APPENDIX A: MECHANICAL DRAWINGS ......................................................................................... 54

APPENDIX B: GLOSSARY .................................................................................................................. 55

APPENDIX C: OTHER INFORMATION ............................................................................................... 56


Dragonfly 500

Revision History

Version

1.0

1.1

1.2

1.3

1.4

1.5

Released

29 Jul 2016

21 Jul 2017

08 Mar 2018

04 Apr 2018

10 May 2018

20 Dec 2018

Description

Initial release - intended for internal use only.

Updated product labels.

Updated cover page design.

Added labelling locations.

Added Camera XY adjustment procedure (Section 4.6.5)

Added references to check MyAndor for further information.

Updated references to Dragonfly as Dragonfly 500. (For Dragonfly 200 refer to the

Dragonfly 200 manual).

Further detail added to Camera XY adjustment procedure (Section 4.6.5)

Information added regarding interlock system (Section 2.5.3)

Updated Camera Zoom image (page 22)

Clarified Cylindrical lens and TIRF modules are not field upgradeable options.

(Section 1.6)

Emission filters replacement procedure is for service personnel only (Section 4.6.3)

Updated laser safety notes for stage cover (Guidelines for Safe Operation of Laser

Products).

U

pdates to the

M

anUal

Changes are periodically made to the product and these will be incorporated into new editions of the manual. Please check for new releases of the manual in MyAndor: http://my.andor.com/login.aspx

. If you find an issue in this manual please contact your customer support representative (Section 1.1) with a description of the issue.


Dragonfly 500

Safety and Warning Information

PLEASE READ THIS INFORMATION FIRST

WARNINGS

• USE OF CONTROLS OR ADJUSTMENTS OR PERFORMANCE OF PROCEDURES OTHER THAN THOSE

SPECIFIED HEREIN MAY RESULT IN HAZARDOUS RADIATION EXPOSURE.

• NOTE THE HAND CRUSH HAZARDS IN SECTION 4.6

1. If the equipment is used in a manner not specified by Andor, the protection provided by the equipment may be impaired.

2. Do not position this product so that it is difficult to operate the mains disconnecting device. See SECTION 4.1,

“Emergency Mains Disconnection” .

3. Before using the system, please follow and adhere to all warnings, safety, manual handling and operating instructions located either on the product, or this manual.

4. Keep this manual in a safe place for future reference.

5. Users must be authorised and trained personnel only; otherwise this may result in personal injury, and/ or equipment damage and impaired system performance.

6. There are no user-serviceable parts inside the product and the enclosure must not be opened. Only authorised service personnel may service this equipment.

7. This product will be used with lasers.

8. IEC Technical Document IEC TR 60825-14 recommends the presence of a Laser Safety Officer (LSO); however, national guidelines should be referred to.

9. This equipment has not been designed and manufactured for the medical diagnosis of patients.

10. Do not attempt to bypass any safety interlocks. They are provided to comply with the safety requirements of various regulatory agencies and must be employed to protect the operator.

11. Protective earth is an integral part of the protection against electric shock in this product, and is provided via the earth pin of the external power supply. Ensure that this is plugged into the building earth system via the mains socket. Do not tamper with any of the earthing measures.

12. Any AC/DC Power Supply used with this product must meet the requirements specified in Section 7.3

.

13. Only use a mains cord certified to the safety regulations for your region and suitably rated for the mains supply specified in Section 7.3

.

14. Make sure the power supply cord is located so that it will not be subject to damage. If replacement of the detachable power supply cord is required, ensure replacement is of same type and rating.

15. Locate the laser source, Dragonfly 500 and any other components connected via fibre optic cables within 2 m of each other so that the interconnecting fibres are not stretched, bent, or constrained.

16. While running an experiment, keep room temperature as stable as possible.


Dragonfly 500

17. Performance of the system may be adversely affected by rapidly changing environmental conditions or operation outside of the operating conditions specified in SECTION 7 “TECHNICAL SPECIFICATIONS” .

18. Electromagnetic Compatibility: This is a Class A product. In a domestic environment this product may cause electromagnetic interference, in which case the user may be required to take adequate measures.

19. This product has been designed and tested to perform successfully in a normal (basic) electromagnetic environment, e.g. a typical life science test laboratory, as per the EU EMC Directive. It is not designed to operate in a harsh electromagnetic environment, e.g. close to the following equipment: EMI/RFI generators, electrostatic field generators, electromagnetic or radioactive devices, plasma sources, arc welders, x-ray instruments, intense pulsed sources, or other similar sources of high energy fields whose emissions are not within the normal range expected under the EU EMC Directive.

20. The green/yellow cable (“Earth Strap”) connecting the optical breadboard to earth is necessary for this product to protect against certain EMC disturbances and to comply with the EU EMC Directive and other international

EMC regulations. It is also a good idea to de-risk such a large piece of uninsulated metal from causing electrocution in the event of damage to mains cables. Do not remove it.

21. Please note that this product is not designed to provide protection from ionising radiation. Any customer using this product in such an application should provide their own protection.

22. Service must only be completed by authorised service personnel with the power turned off as internal moving parts are not interlocked and should be considered potential pinch points.

23. This product is a precision scientific instrument containing fragile components. Always handle it with care.

24. Ensure fibre cables are not bent tightly as this may damage the internal optical fibre. For transport or storage, fibre cables should be coiled with a diameter of 300 mm or greater.

25. Do not wet or spill liquids on the product, and do not store or place liquids on the product.

26. If spillage occurs on the product, switch off power immediately, and wipe off with a dry, lint-free cloth.

27. If any ingress of liquids has occurred or is suspected, unplug the mains cables, do not use, and contact Andor customer support.

28. Do not expose the product to extreme hot or cold temperatures.

29. Do not expose the product to open flames.

30. Do not allow objects to fall on the product.

31. See SECTION 5.1, “Cleaning and Decontamination” .


Dragonfly 500 l

aser

s

afety

WARNING – USE OF CONTROLS OR ADJUSTMENTS OR PERFORMANCE OF PROCEDURES OTHER THAN

THOSE SPECIFIED HEREIN MAY RESULT IN HAZARDOUS RADIATION EXPOSURE

This product is designed to be integrated with lasers that cover the entire visible spectrum and extend into the invisible infra-red spectrum. This section of the manual is designed to make the end-user aware of the hazards of the product due to these lasers. Laser safety hazards differ from those of normal light sources and users must be familiar with the hazardous properties of lasers as these are high intensity, highly collimated beams of electromagnetic radiation.

Andor recommend that all facilities have an established system for the safe use of lasers as per their national frameworks and Occupational Health and Safety legislation. “IEC 60285-1” and “American National Standard Z136.1-

2007- Safe use of Lasers” may be useful references for good practice.

l

aser

p

rodUct

c

lassification

The Dragonfly 500 does not contain any embedded laser sources but it is intended to be integrated with a compatible laser source and has therefore been designed to safely accept, and handle laser radiation. The Dragonfly 500 unit and all ancillary components of the Dragonfly 500 system have been designed and labelled to be compliant with IEC 60825-1 and CDRH Title 21 CFR 1040.10.

When assembled and integrated in the intended manner, Dragonfly 500 will form part of a unique laser product formed by the entire system. The classification of this product will depend entirely on the laser source connected and the associated research-grade microscope. Therefore the final system must be assessed, classified and labelled in accordance with the requirements of IEC 60825-1 and CDRH Title 21 CFR 1040.10.

This must be performed by a competent, trained individual e.g. the site’s Laser Safety Officer. In such cases where the system is installed by an Andor representative, it is Andor’s responsibility to assess and classify the final system.

The customer support engineer will be competent and trained to perform this and ensure that the system is correctly classified and labelled (this is shown later in this Section “Laser Classification for an installed system”).

The customer support engineer will be competent and trained to perform this and ensure that the system is correctly classified and labelled. Please refer to Section 3.5

for guidance on the assessment and classification procedure.


Dragonfly 500 l

aser

s

afety and

p

rodUct

c

oMpliance

l

abels

All relevant safety compliance information is visibly displayed on the product as per the International Laser Safety

Standard IEC 60825-1 and the U.S. Laser Product Performance Standard 21 CFR 1040.10.

As the Dragonfly 500 system will form part of a unique configuration of components, it cannot be classified in isolation.

Therefore when integrated, it may form part of a system with one of the classifications described below. A label in the format depicted below will be included for application by the system integrator following assessment and classification.

The Dragonfly 500 may be configured as part of a larger system that includes multiple Class 3B and Class 4 lasers. A number of separate emission wavelengths can also be available in any one system. In some very rare occasions another classification label may be used, this will be explained in additional documentation if this is required.

Class 3B

Laser products that are normally hazardous when intrabeam ocular exposure occurs

(i.e. within the Nominal Ocular Hazard Distance, NOHD) including accidental short time exposure. Viewing diffuse reflections is normally safe. Class 3B lasers which approach the

Accessible Emission Limits (AEL) for Class 3B may produce minor skin injuries or even pose a risk of igniting flammable materials. However, this is only likely if the beam is a small focused spot.

Note: There exist some theoretical (but rare) viewing conditions where viewing a diffuse reflection could exceed the Maximum Permitted Exposure (MPE). For example for Class

3B lasers having powers approaching the AEL, lengthy viewing of greater than 10 s of true diffuse reflections of visible radiation and viewing at distances less than 13 cm between the diffusing surface and the cornea can exceed the MPE.

Class 4

Laser products for which intrabeam viewing and skin exposure is hazardous and for which the viewing of diffuse reflections may be hazardous. These lasers also often represent a fire hazard.


Dragonfly 500 l

aser

c

lassification

l

ocations

The access panels are labelled with Laser Classification / Explanatory Labels- as described later in this section. If the

Dragonfly 500 is to be re-classified by a site’s trained and competent individual (e.g. the site’s Laser Safety Officer) then they must ensure that labels of the correct classification and wavelength range are placed in all of the locations.

l

aser

a

pertUre

The System Integrator MUST ensure that the final system’s laser aperture is suitably labelled e.g. the microscope’s

Objective is identified by a label on the microscope’s stage top.

The appropriate aperture label(s) are included with the documentation for application by the system integrator. The location and example labels are shown later in this section.

d

escription of

e

Mitted

r

adiation froM the

l

aser

p

rodUct

Parameter

Wavelengths

Beam Divergence

Maximum Power or

Energy Output

Pulse Duration

Values

400-800 nm

0.3-1.49 NA

<1W

N/A

Notes

Exact outputs will depend on the integrated laser source, please refer to it’s Explanatory Label and User Documentation

Exact divergence will be dependent on the objective in use, please refer to the microscope’s User Documentation

The maximum output power will depend on the integrated laser source and the configured optical elements but will be significantly less than 1W (Class 4)

All recommended laser sources for the Dragonfly 500 are Continuous

Wave output

All recommended laser sources for the Dragonfly 500 are Continuous

Wave output

Pulse Repetition

Rate

N/A

Irregular Pulse

Pattern

N/A All recommended laser sources for the Dragonfly 500 are Continuous

Wave output d

escription of

e

Mitted

r

adiation froM the

i

ntegrated

l

aser

s

oUrce

This is dependent on the laser source integrated with the Dragonfly 500 system. As there are multiple options that are available from Andor, or the Dragonfly 500 may be integrated with a customer’s existing laser source(s), please refer to the laser source’s explanatory label and user documentation for a description of its emitted radiation.

r

ecoMMended

r

esponsibilities of a

l

aser

s

afety

o

fficer

These include, but are not restricted to, the following; however, national guidelines should also be referred to:

1. Ensure that all personnel requiring access to the product are fully trained in both using the product and the general use of Class 3B and Class 4 lasers (see below).

2. Ensure users are familiar with the hazardous properties of lasers; namely that laser safety hazards differ from those of normal light/radiation sources as they are high-intensity, highly collimated beams of electromagnetic radiation.

3. Ensure that equipment is used in a controlled area by trained end users in accordance with national guidelines.

4. Ensure end-users are familiar with the operation of the laser’s key switch control, interlocks, emission LEDs and other safety features.

5. Ensure that all interlocks are connected and functioning correctly (see Sections, 2.5.1 and 2.5.3

).

6. We recommend that a copy of IEC 60285-1 is purchased by the laser safety officer for reference.


Dragonfly 500 g

Uidelines for

s

afe

o

peration of

l

aser

p

rodUcts

1. Read the safety instructions supplied with all equipment in the system.

2. Never look into a laser beam either directly, or indirectly.

3. Do not attempt to disassemble the unit housing the lasers or any part of the system. If there a problem is suspected, please contact Andor directly (see Section1.1, “Help and Technical Support” ).

4. Restrict and control access to the area(s) where laser(s) are in use to those persons who are trained in the dangers of lasers and trained on the safety precautions to be observed when working with lasers.

5. Ensure suitable laser warning signs are prominently displayed in the area the system operates.

6. If the system is not in use turn the laser off using the key switch.

7. On a daily basis, or before every use, verify that the laser interlock circuit ( Section 2.5.3

) is working by confirming that the laser emission indicator on the source turns off when either of the following are done:

• The microscope binocular eyepieces are in the open position.

• The articulated transmitted light arm on inverted microscopes is tilted back from the functional vertical position before using the system.

• The laser safety stage cover or protective shield is opened.

8. If it is necessary to visually locate and indicate the output of invisible wavelengths, laser viewing cards for this purpose e.g. Thorlabs NIR Detector Card VCR5 (for 700- 1400 nm) must be used along with the appropriate laser safety goggles.

9. Additional precautions may need to be implemented as the necessary precautions will be specific to each system installation, configuration and typical mode of use. The responsible Laser Safety Officer must assess and implement the necessary precautions to avoid possible exposure to hazardous radiation during use.

e

ye

p

rotection

Eye protection is not required for the safe use of the device as the only radiation observable (without intentional misuse) is directionally stable, diffuse and highly divergent from the designated aperture which is static and labelled. Eye protection must be used by all Installation and service personnel when accessing any radiation during any installation or service procedure.

If eye protection is deemed desirable by the local Laser Safety Officer, Andor recommends the following products:

• 360 nm – 510 nm – Kentek KXP-4001 Spectacles

• 510 nm – 670 nm – Kentek KRA 6702 Spectacles

• 670 nm – 800 nm – Kentek KMZ-215C Spectacles


Dragonfly 500 i

nforMation on

V

iewing

a

pertUres

In order to view any of the apertures, it is necessary to use eye protection to reduce the intensity of the laser power to acceptable levels for viewing. The table below provides the appropriate OD values required for each wavelength.

WARNING: THE USER IS RESPONSIBLE FOR ACQUIRING THE APPROPRIATE GOGGLES TO MEET THESE SAFETY

REQUIREMENTS IF VIEWING IS REQUIRED.

It should also be noted that since the laser selection covers many wavelengths, it may not be possible to find a single pair of goggles to provide the necessary protection.

Wavelength (nm)

514

515

532

552

561

594

405

445

458

473

488

637

640

685

730

750

785

Max. Laser Power (mW)

36

135

90

90

135

45

180

67.5

67.5

67.5

180

126

126

126

27

360

108

Optical Density for Safe Viewing

3

3

3

3

3

3

4

4

4

4

4

3

4

3

3

3

3 w

orking with

o

ptical

f

ibres

1. Only service personnel authorized by Andor should remove or inspect fibres.

2. The laser radiation passing through fibres is potentially hazardous, so great care should be taken to avoid exposure to this radiation.

3. The fibre can be easily damaged by bending or general mishandling. Ensure that the minimum curvature is never exceeded when handling. Recommended minimum bend radius is 300 mm.

4. Optical fibres are prone to damage by bending local to the connector.

5. The coupler is not designed to withstand pulling of the fibre. If the fibre is pulled the system performance could be compromised or the system may fail.


Dragonfly 500

PRODUCT INFORMATION AND WARNING LABELLING

This section highlights the location and description of the product information and warning labels of the Dragonfly 500.

P roduct

c omPliance and

i nformation

l abels

Connections

Panel Label

(see Section 2.5.1)

Standby Button

Label

(see Section 2.5.2)

Figure 1: Product Compliance and Information Label (Connections Panel and Standby Button labels also shown)

This label indicates the product compliance with the applicable standards. It also provides information on the manufacturer and identifies the model, serial number, and manufacturing date (month, year). A warning also highlights that the Dragonfly 500 requires a multi-person lift for transport and installation (refer to Section 3 ).

i nner

d oor

l abels

Figure 2: Inner Door Labelling

This label located on the Emission Pathway Panel shows the opening instructions e.g. to access the filter wheels.


Dragonfly 500

P ort

s election

m irror

a ccess

l abels

Information for each dichroic installed is listed- as shown in the above example.

Figure 3: The Port Selection Mirror Access Label

This label highlights the potential Dichroic Access Door Crush Hazard and information for the installed Dichroics. Power

Off the Dragonfly 500 before accessing the Camera Splitter dichroic slider. Please see Section 4.6

.

f ilter

W heel

l abels

Each filter wheel is labelled with information of each filter installed (as shown in the above example)

Note the hand crush hazard warning for each filter wheel. Do not place any limb or item into the Filter

Wheel aperture when a filter wheel is removed.

Figure 4: Location of Filter Wheel Labels

Further information on the exchange of filters and dichroics is shown in Section 4.6

.


Dragonfly 500 s ervice

a ccess

P anel

l abels

Note: Units without TIRF module also have warning on the blanking panel

(requires tooled removal)

Figure 5: Panel Access Laser Warning Labels

The above label highlights the accessible laser hazard (in this example Laser Class 3B) if the panels are removed.

The above label located under the Engine

Access Panel highlights the potential hand crush hazard if the top panel is removed.

THIS PANEL MUST NOT BE REMOVED BY THE

USER.

Figure 6: Engine Access Top Panel (Service only) l aser

a

Perture

W arning

l abels

(m icroscoPe

o culars and

s tage

)

Version 1.5 rev 20 Dec 2018

Figure 7: Microscope Laser Hazard warning labels on Oculars (left) and Stage (right)

15

Dragonfly 500

INTRODUCTION

SECTION 1: INTRODUCTION

1.1 t

echnical

s

Upport

If you have any questions regarding the use of this equipment, please contact the representative* from whom your system was purchased, or:

Europe

Andor Technology Ltd.

7 Millennium Way

Springvale Business Park

Belfast

BT12 7AL

Northern Ireland

Tel. +44 (0) 28 9023 7126

Fax. +44 (0) 28 9031 0792

USA

Andor Technology

425 Sullivan Avenue

Suite # 3

South Windsor

CT 06074

USA

Tel. +1 (860) 290-9211

Fax. +1 (860) 290-9566

Asia-Pacific

Andor Technology (Japan)

IS Building

3-32-42 Higashi-Shinagawa

Shinagawa-ku,

Tokyo 140-0002

Japan

Tel: +81-3-6732-8968

Fax: +81-3-6732-8939

China

Andor Technology

Unit 1, Building A,

66 Zhufang Rd,

Haidian Dist.

Beijing 100085

China

Tel: +86 (0)10 8271 9066

Fax. +86(0)10 8271 9055

* The latest contact details for your local representative can be found on the contact and support page of our website.


Dragonfly 500

INTRODUCTION

1.2 d

isclaiMer

THE INFORMATION CONTAINED HEREIN IS PROVIDED “AS IS” WITHOUT WARRANTY, CONDITION OR

REPRESENTATION OF ANY KIND, EITHER EXPRESS, IMPLIED, STATUTORY OR OTHERWISE, INCLUDING BUT NOT

LIMITED TO, ANY WARRANTY OF MERCHANTABILITY, NON-INFRINGEMENT OR FITNESS FOR A PARTICULAR

PURPOSE.

IN NO EVENT SHALL ANDOR BE LIABLE FOR ANY LOSS OR DAMAGE, WHETHER DIRECT, INDIRECT, SPECIAL,

INCIDENTAL, CONSEQUENTIAL OR OTHERWISE HOWSOEVER CAUSED WHETHER ARISING IN CONTRACT, TORT

OR OTHERWISE, ARISING OUT OF OR IN CONNECTION WITH THE USE OF THE INFORMATION PROVIDED HEREIN.

1.3 c

opyright and

p

rotectiVe

n

otices

The copyright in this document and the associated drawings are the property of Andor Technology Ltd. and all rights are reserved. This document and the associated drawings are issued on condition that they are not copied, reprinted or reproduced, nor their contents disclosed.

The publication of information in this documentation does not imply freedom from any patent or proprietary right of

Andor Technology Ltd. or any third party.

1.4 t

radeMarks and

p

atent

i

nforMation

Andor and the Andor logo are trademarks of Andor Technology Ltd. Andor Technology Ltd. is an Oxford Instruments company. All other marks are property of their owners. Borealis includes technology covered by the following patents:

US Patent No. 8,275,226, EP Patent No. 2196839, US Patent No. 8,670,178 and CA Patent No. 2779146.


Dragonfly 500

INTRODUCTION

1.5 s

Upplied

c

oMponents

The Dragonfly 500 system may be supplied in different configurations along with components and accessories required for your specific application. Please refer to your ordering information and check the packing list to confirm that all parts are present.

Description

Quantity

Dragonfly 500 Unit

(note configuration and functionality depends on Dragonfly model ordered)

1

24 V Power Supply

Power cable (country specific)

User manuals in electronic format 1

PC WorkStation (as ordered)

Regional keyboard

Software (as ordered e.g. Fusion) -

Filter Wheel, 8 position, 25 mm (inc. cover plates and 1” filter blanks)

2*

Male-male null-modem DB-9 RS-

232 serial cable (supplied with filter wheel controller)

1

FW-1000 female-female DB-9 drive cables (supplied with filter wheels)

2

Notes : Filters as specified at time of ordering.

1

1

1

1


Emission filter wheel controller

Microscope port adapter and bellows (microscope specific, as ordered)

USB 2.0 male-male A-B cable

(locking type)

Borealis Beam Conditioning Unit

1

1

1

1

Multi-mode optical fibre, 2 m, 50 µm 1

Single-mode optical fibre

(as ordered for TIRF)

1

Interlock kit 1

Metric (M6) and imperial (1/4”-20) feet locking screws

3 of each type

Microscope raising pedestal kit

(microscope specific, as ordered)

1

18

Dragonfly 500

INTRODUCTION

1.6 o

ptional

c

oMponents

There are a range of optional components available for the Dragonfly 500 including:

1. Multi-band excitation dichroic optics for multiplexing excitation laser light (up to 3 optics, motorized selection, installed by service/support engineer only).

2. Camera Port Selection/Image Splitting dichroic Broadband mirror/window for 100% throughput camera port selection or dichroic mirrors for simultaneous emission splitting (up to 4 filters per cassette, motorized selection and user exchangeable).

3. Emission filters (8 per wheel, motorized selection and user exchangeable).

4. Additional motorized emission filter wheels (user exchangeable).

5. Cylindrical lens for 3D super-resolution localization microscopy or particle tracking applications (must be specified at time of order, not field-upgradeable).

6. Imaris Software for advanced image analysis and visualisation capabilities.

7. TIRF Module can be fitted to upgrade models without TIRF (must be specified at time of order, not fieldupgradeable).

Notes:

• Andor provide a range of other microscopy system components that can be configured with or alongside

Dragonfly 500. Please contact your nearest Andor Sales representative for further information on these other system accessories.

• Use only accessories supplied and recommended by Andor (except for the AC/DC Power Supply, which must meet the specification of Section 7.3

, and mains cables, which must be certified for the country of use).

1.7 r

ecoMMended

e

Mission

f

ilters

For the latest information on available emission filter options please visit the Dragonfly page on andor.com

.

Please contact your nearest Andor Sales representative for further information on these and other system components available.


Dragonfly 500

PRODUCT OVERVIEW

SECTION 2: PRODUCT OVERVIEW

This section provides an overview of the main features of the Dragonfly 500.

2.1 a

n

i

ntrodUction to the

d

ragonfly

The Dragonfly 500 is a multi-modal imaging platform enabling laser-based widefield , spinning-disk confocal , and optionally Total Internal Reflectance Fluorescence (TIRF) microscopy when connected to a research-grade microscope. This product is intended for professional scientific research applications, especially live-cell biological imaging and photo-stimulation.

1

2

Figure 8: The Dragonfly 500- shown with Andor iXon Ultra EMCCD and Zyla sCMOS Cameras

2.1.1 s

ysteM

i

ntegration

The Dragonfly 500 is shown as part of an integrated system in the diagram of Figure 9. Laser light from an Integrated

Laser Engine (ILE) is fed as an input into one port of the Dragonfly 500 via a multi-mode optical fibre for widefield and confocal illumination (passing first through a Borealis Beam Conditioning Unit – BCU ) and/or a single-mode optical fibre through a different port for TIRF illumination.

Camera

Single-mode fibre TIRF Input

Filter Wheel Controller

Multi-mode fibre

BCU Output

Widefield/

Confocal

Input

Camera

Microscope

ILE BCU

Dragonfly 500

Figure 9: A Simple Representation of a Dragonfly 500 System (with ILE, BCU and Cameras)


Dragonfly 500

PRODUCT OVERVIEW

The Dragonfly 500 couples to an inverted microscope through its left side port using a microscope-specific connector flange and mating optics. Proper and secure mating of the Dragonfly 500 to a microscope requires that the microscope be raised to match the optical height of the Dragonfly 500 using the supplied pedestal kit and fastened onto an optical table benchtop having regularly spaced metric M6 or imperial ¼”-20 screw holes. The minimum recommended table benchtop size is 122 x 92 cm [48 x 36 inches] approx. For laser safety reasons, the Dragonfly 500 should only be aligned and interlocked to the microscope by a qualified Andor representative.

Illumination laser light transferred through the Dragonfly 500 enters the microscope’s left side port, and subsequent fluorescence emission light generated by the sample specimen and collected by the microscope optics returns to the

Dragonfly 500 through the same port. The emission light is separated from the excitation laser light by a multi-band excitation dichroic optic and forms an image on the camera after passing through an appropriate interference filter loaded in a motorized filter wheel.

2.1.2 o

Utline of the

f

Unctionality of

i

Maging

M

odes

*

Multi-Mode Imaging - Different fluorescence imaging modes (Widefield, Confocal, and TIRF) are selected by changing the ILE (laser engine) output line and the position of the confocal disk in the imaging path. The microscopic specimen is viewed through various imaging “channels” defined through different combinations of laser wavelengths, excitation dichroic optics, and emission filters. Conventional transmitted white light imaging techniques such as brightfield and differential interference contrast (DIC) can be used when the Dragonfly 500 is set to the Widefield imaging mode with none of the ILE lasers active. In this state, the Dragonfly 500 acts as an image relay to the camera. Localisation

Microscopy introduces a cylindrical lens pair into the widefield imaging path to facilitate enhanced resolution 3D localisation methodologies.

Dual Pinhole size* - The confocal disk possesses two distinct pinhole sizes (25 µm and 40 µm) which can be selected to vary the degree of optical sectioning.

Muti-mode, common optical pathway - A unique feature of the Dragonfly 500 is that the widefield/confocal and

TIRF illumination are combined into a common optical pathway permitting images from all three fluorescence imaging modalities to be captured by the same camera in registration with each other. Transmitted light images (brightfield, DIC, etc.) are also captured in registry with these fluorescence images.

Illumination Zoom and Variable Illumination Field Aperture Size - The Dragonfly 500 possesses the ability to change the size of the widefield/confocal illumination field to optimally fill the active area of detector camera sensor or to purposefully illuminate a sub-region of the sensor area for applications that require localized illumination within the field of view at higher laser power densities. Ten different illumination sizes (apertures) are available.

Pos 1: 16.9 x 14.3

Pos 2: 14.9 x 14.3

Pos 10: 3.6 x 3.6

Pos 9: 4.4 x 4.4

Pos 3: 13.0 x 13.6

Pos 8: 4.9 x 3.7

Pos 7: 7.0 x 7.0

Pos 6: 7.4 x 7.4

Pos 5: 8.5 x 8.5

Pos 4: 9.1 x 7.4

The size of TIRF illumination field is fixed and slightly overfills the sensor area of Andor iXon Ultra 888 EMCCD or Zyla 4.2 sCMOS cameras. Depending on the Dragonfly 500 model, four different illumination zooms are available in widefield mode and two in confocal mode. NOTE: Absolute Magnification factors are approximate and will vary depending on microscope model. Calibration via software is required for accurate measurements.

Figure 10: Changing the illumination field aperture size.

* note Imaging modes and functionality will depend on Dragonfly model, see Section 2.2

.


Dragonfly 500

PRODUCT OVERVIEW

Relative illum.

Power Density

1x

1x

1x

2x

2x

2x

2x

2x

2x

2x

2x

2x

2x

Camera

Magnification

(nominal)

1x

1x

1x

1x

1x

2x

1x

2x

2x

2x

1x

2x

2x

Field of view

(mm)

16.64x14.04

14.04x14.04

13.3x13.3

8.8x7.1

8.2x8.2

14.04x14.04

6.7x6.7

13.3x13.3

13.3x13.3

9.1x6.8

4.1x4.1

8.2x8.2

6.5x6.5

Comments

Neo&Zyla 5.5 (2560x2160 sCMOS) - Full frame

Neo&Zyla 5.5 (2160x2160 sCMOS) – Scope Quality Area

Field Number Limit

Zyla 4.2 (2048x2048) and Ixon 888 sCMOS 1392x1080 optically centred window

(200 fps) iXon 897

Neo&Zyla 5.5 (2160x2160 sCMOS) – effective pixel size

= 3.25µm iXon 888:

512x512 ROI Optically Centred Crop iXon 888:

256x256 ROI Optically Centred Crop

Zyla 4.2 (2048x2048) – effective pixel size = 3.25 µm sCMOS 1392x1080 optically centred window (200 fps):

3.25 or 6.5 µm (binned 2x2 with higher frame rate)

Ixon 897:

256x256 ROI Optically Centred Crop

Works for iXon 897 – effective pixel size = pixel 8 µm iXon 888 quadrant crop mode effective pixel size = 6.5 µm

Min Disk Area Illum.

Dimensions (mm)

16.64x14.04

14.04x14.04

13.3x13.3

8.8x7.1

8.2x8.2

7.1x7.1

6.7x6.7

4.6x3.4

4.1x4.1

3.25x3.25

Camera Zoom - The additional magnification provided by the zoom lenses can be used to alter the spatial sampling frequency of the microscope image by the camera, which may be desirable for applications such as super-resolution localization microscopy or particle tracking.


Figure 11: Camera Zoom

22

Dragonfly 500

PRODUCT OVERVIEW

Dual Detector Ports - The Dragonfly 500 can be equipped with one or two camera detectors. Each camera port has its own 8-position filter wheel and three selectable zoom lenses that offer 1x, 1.5x, or 2.0x additional magnification of the microscope image.

Emission Filter and Optical Splitter Flexibility - The emission light from the microscope may be completely directed to one of these camera ports via a 100% broadband transmitting window or a 100% broadband mirror. Alternatively, the emission light can be spectrally divided simultaneously between the two camera ports with an emission splitting dichroic mirror. The Dragonfly 500 can be loaded with up to four distinct image splitting optics.

The emission filter wheels (as well as their filters) and image splitting optics are accessible to the user through the interlocked front panel of the Dragonfly 500. These components can be safely removed and exchanged/swapped with additional filters/filter wheels and/or image splitting optics (see Section 4.6.1) . Each supplied filter wheel has an RFID tag which contains the details of the loaded emission filters so when loaded the control software will automatically pick up the new filter wheel information. The RIFD Tag can be updated by the User via the Dragonfly 500 Control software if the emission filters loaded in a Filter Wheel are changed.

2.2 s

ysteM

c

onfigUrations

The Dragonfly 500 is available in different configurations that feature the following functionality:

Description

Borealis illumination

NIR Extended Range

Laser and Widefield illumination

Laser and Widefield imaging

3-position motorised camera magnification changer (1x, 1.5x, 2x)

Zoom illumination

40 m m Confocal Disk pattern

25 m m Confocal Disk pattern

2 nd camera port with 4 position splitter

TIRF imaging (simultaneous multicolour)

Astigmatic Lens (3D localisation)

• Standard o

Option (Field or factory fitted o Option (Factory Fitted)

301

•

•

•

•

•

• o

302

•

•

•

•

•

• o o

•

•

•

•

•

502

•

•

• o o o

•

•

•

•

503

•

•

• o o

•

•

•

•

•

•

505

•

•

•

•

•

•

•

2.3 M

icroscope

r

eqUireMents

The mating optics and mechanical hardware that couple the Dragonfly 500 to the microscope are specific to the microscope model, objective lens, and any other system component that possibly changes the spacing between the tube lens and the objective lens back-focal plane (number of decks - single vs. double deck, stage model, stage risers, incubation chambers, objective piezo focus drive or autofocus modules). Therefore, it is essential that these components be specified correctly at the time of ordering. The Dragonfly 500 couples to an inverted microscope through its left side port using a microscopespecific connector flange and mating optics. The system microscope must possess a 100% left side imaging port selection option. Partial image port splitting optics (80/20 port selection option for example) are discouraged, and if present must be properly interlocked to the laser source by the system installer (refer also to Section 2.5.3).

2.4 c

aMera

o

rientation

r

eqUireMents

Each camera must be oriented/rotated on each camera port such that the image as viewed in the image acquisition software matches the orientation of the image when viewed through the microscope eyepieces. This is configured during installation and should not normally need adjustment. If this is required, please contact your Andor customer support representative for further information.


Dragonfly 500

PRODUCT OVERVIEW

2.5 e

xternal

f

eatUres of the

d

ragonfly

500

The main external features and modules of the Dragonfly 500 are shown below- with 2 cameras mounted to the side and top camera ports (Installation handles are removed after installation).

Fibre Ports

Camera Port 1

Horizontal

Microscope

Port

Front Access Panel

(pull top left corner forward to open)

Figure 12: Dragonfly 500- Top View

Camera Port 2

Vertical

Front Access Panel

Connections

Panel

Standby button

Lifting Handle

(for installation)

Lifting Handle

(for installation)

Figure 13: Dragonfly 500- Left Side View

WARNING: Access points on the Dragonfly 500 (shown below) are marked for service access only and must not be opened by the user.

Figure 14: Factory Only Access Labels- For Service Access Only.


Dragonfly 500

PRODUCT OVERVIEW

2.5.1 c

onnections

p

anel and

s

tatUs

d

isplay

The main connections panel is located on the left side panel (refer also to figure 13).

On/Off

Function

FW1 In

FW2 In

RS232

(FW out and PC)

External

Control

USB

System Interlock

Sync Out

Sync In

Power 24V

Figure 15: Connection Panel

On/Off

Turn the Dragonfly 500 system ON and OFF.

Power 24V

See Section 7.3

for specification.

USB

The USB port connects the Dragonfly 500 to a PC workstation over a USB link allowing the Dragonfly 500 to be fully controlled through the image acquisition control software on the PC (do not connect RS232 if controlling via USB).

System Interlock

The Interlock System output ( Section 2.5.3

) provides a set of volt free normally closed contacts that are connected to the user’s laser safety interlock. If any of Dragonfly’s access panels are removed, the closed contacts open, breaking the circuit.

Sync Out

Sync out provides a TTL level synchronising clock output signal based on the confocal disk rotation. Every time the disk completes a revolution a 10 m s wide logic high pulse is generated at this output. This pulse can be used to synchronise external equipment to the disk rotation.

Sync In

Sync In is used to synchronise the confocal spinning disk to an external timing signal. The timing signal needs to be TTL level (currently not enabled).

FW1 In

This port is connected to the port marked W1 on the filter wheel controller with the supplied cable. This allows filter wheel 1 to be controlled.


Dragonfly 500

PRODUCT OVERVIEW

FW2 In

This port is connected to the port marked W2 on the filter wheel controller with the supplied cable. This allows filter wheel 2 to be controlled.

RS232 FW out

This port is connected to the port marked RS-232 on the filter wheel controller to allow Dragonfly 500 to effect control of the filter wheels.

RS232 PC

This port connects the Dragonfly 500 to a PC workstation over an RS232 link allowing Dragonfly 500 to be fully controlled from the PC (do not connect USB if controlling via RS232).

External Control

This port gives the option of controlling Dragonfly 500 without using a PC. This option is currently not enabled.

Function

Pressing this button will cause Dragonfly 500 to enter a pre-set configuration.

2.5.2 p

ower

s

Upply

The external power supply requirements are outlined below:

Parameter

External Power Supply

Power Supply output

Power Supply Output Connector

Value

100-240 VAC, 50-60 Hz

24 VDC, 5 A Max*

Fischer S104 A037-230+ (shielded cable connector)

E3 104.3/7.7 + B (clamp set)

Pins 1, 2 +24V,

Pins 3, 4 0V

P oWer

s tatus

i ndication

Figure 16: Power Supply Output Connector (Fischer S 104 A037-230+)

Figure 17: Power button with system status indication

Status

OFF

Orange

Green

Power off

Standby Going into Standby, coming out of Standby (including initialising from power on)

Active i.e. not in Standby


Dragonfly 500

PRODUCT OVERVIEW

2.5.3 l

aser

s

afety

i

nterlock

s

ysteM

The Dragonfly 500 is interlocked as part of a laser safety interlock system with external equipment such as a laser source, microscope or door interlock. The System Interlock connection of the Dragonfly 500 is located on the connection panel (See Section 2.2.1) . The internal interlock system of the Dragonfly 500 consists of switches in the main filter wheel internal door (not the external cosmetic panel) and switches in the Disk Dichroic Access Panel (service access only). When either door or panel is opened, the external laser sources interlock system will be open-circuited, causing the laser to turn off.

Figure 18: Access Warning Label (on main filter wheel internal door).

PLEASE NOTE: It is important that the interlock system assembled by the system installer is not disassembled as this could affect your ability to use the laser or could create a laser safety risk.

For Class 3B : the laser configuration is re-established to what it was, once the interlock is closed again.

For Class 4 : the shutters of the ILE must be re-opened to regain the previous laser configuration. Alternatively, the Key

Switch of the ILE may be cycled OFF and then ON again to restart the ILE. However, in this case, the laser configuration must be re-established entirely.

Figure 19: The status of the ILE Laser combiner can be viewed from the Emission Status indicator lights.

Left: Emission lights off - no laser emission is possible.

Right: Emission lights on - laser emission possible.


Dragonfly 500

INSTALLATION

2.5.3.1 i

nterlock

h

igh

-l

eVel

s

UMMary

d

iagraM

Laser Source:

e.g.

• ILE Laser

Combiner with

Borealis

Notes

The Dragonfly features internal interlocks (Section 2.5.3) and the interlock system is extended to the other connected devices. If any of the interlocks is activated laser emission will cease.

Dragonfly

Internal interlocks activate to cease laser emission if the internal door or panels are removed.

Microscope

Microscope may feature a number of interlocked components including:

• Arm Interlock

• Turret interlock (where applicable)

• Eye interlock

Laser emission will cease if any of the interlocks are activated.


Misc. Components

Additional components may form part of the interlock system:

• Door Interlock

• Other devices e.g. Mosaic or

MicroPoint

28

Dragonfly 500

INSTALLATION

SECTION 3: INSTALLATION

WARNINGS:

• THE DRAGONFLY SYSTEM MUST BE INSTALLED BY AN AUTHORIZED INSTALLATION ENGINEER

ACCORDING TO THE INFORMATION PROVIDED BY ANDOR.

• THE DRAGONFLY WEIGHS OVER 60 KGS (132 LBS). DUE CARE MUST BE TAKEN WHEN LIFTING THE

DRAGONFLY AS OUTLINED IN THE ANDOR INSTALLATION AND SERVICE MANUAL.

• ONCE INSTALLED AND SECURED TO THE OPTICAL BENCH IT SHOULD NOT BE NECESSARY TO MOVE OR

ADJUST THE MOUNTING OF THE DRAGONFLY OR OTHER COMPONENTS. IF THIS IS REQUIRED, PLEASE

CONTACT YOUR ANDOR REPRESENTATIVE FOR FURTHER INFORMATION.

• ANY INSTALLATION STEPS INVOLVING LASERS MUST BE PERFORMED BY QUALIFIED PERSONNEL USING

PERTINENT LASER SAFETY PROTOCOLS

• SETUP AND OPERATION OF OTHER SYSTEM COMPONENTS ARE DESCRIBED IN THEIR RESPECTIVE

GUIDES AND MANUALS.

3.1 l

ocation and

M

oUnting

• Temperature and humidity must meet the specifications defined in SECTION 7 .

• Operational vibrations should be reduced as much as possible for stability of the imaging train.

• It is recommended that the Dragonfly 500 is mounted on a damped optical table (minimum dimensions: 122 x 92 cm [48 x 36 inches approx.). Please contact your Andor Representative for assistance in specifying and ordering a suitable table.

• Power cabling and control cables should be routed to prevent accidents, damage and accidental unplugging while avoiding bend radii of less than 30 mm.

3.2 V

entilation

Do not cover equipment during operation - all system devices require cooling of internal components to the surrounding ambient environment.

3.3 a

sseMbly

This product requires no assembly by the end user. It must be installed by an installation engineer authorized by Andor or an approved distributor.


Dragonfly 500

INSTALLATION

3.4 t

ransport and

s

torage

• Keep dry, humidity <70%, non-condensing.

• Store between 0°C to 50°C.

• If the Dragonfly 500 needs to be transported, or moved to a different location please contact Andor customer support for further instructions.

• The Dragonfly 500 has handles to aid lifting and positioning of the unit during installation (shown below). These are removed after installation and must be kept in a safe place in case they are needed at a later time.

Figure 20: Installation handles- store in a safe place in case they are needed at a later time.

3.5 l

aser

c

lassification of an

i

nstalled

s

ysteM

The proposed installation scheme of all systems is captured and assessed for all orders received by Andor. The majority of systems can be safely classified at Factory QC as Class 3B products.

The only case to re-assess the accessible emissions is if the beam path appears to be fully enclosed or if the device is being installed into a non-standard configuration e.g. being installed in conjunction with additional laser based products.

The following test criteria should be applied if a system is felt to be fully enclosed e.g. uses a stage cover and/or environmental enclosure.

Assess if any of the laser emissions are open to human access as defined by:

1. Ability of the human body to meet laser radiation emitted by the laser product, i.e. radiation that can be intercepted outside of the protective housing, or

2. Ability of a cylindrical probe with a diameter of 100 mm and a length of 100 mm to intercept levels of radiation of

Class 3B and below, or

3. Ability of a human hand or arm to intercept levels of radiation above the AEL of Class 3B,

4. Also, for levels of radiation within the protective housing that are equivalent to Class 3B or Class 4, ability of any part of the human body to meet hazardous laser radiation that can be reflected directly by any single introduced flat surface from the interior of the product through any opening in its protective housing.


Dragonfly 500

OPERATION

The standard Ophir PD-300W-W sensor and power meter can be used to assess accessible power levels in conjunction with suitable safety glasses and applicable Safe Systems of Work. Any queries should be referred to the Andor Product

Laser Safety Officer for guidance.

If human access is not possible then the device should be reclassified as a Class 1 device . This involves the following steps:

1. Remove all other explanatory (classification) labels EXCEPT those on the laser sources (these count as Removable

Laser Sources and need to remain labelled and classified as stand-alone products).

2. Fit a Class 1 explanatory label onto a permanently affixed surface which is easily visible before and during operation of the system.

3. Ensure a Laser Hazard Symbol is clearly visible before and during operation and affix a label if not.


Dragonfly 500

OPERATION

SECTION 4: OPERATION

WARNINGS:

• IF THE EQUIPMENT IS USED IN A MANNER NOT SPECIFIED BY ANDOR SYSTEM DISTRIBUTORS THE

PROTECTION PROVIDED BY THE EQUIPMENT MAY BE IMPAIRED.

• READ THE USER GUIDES SUPPLIED WITH YOUR SYSTEM COMPONENTS AND SOFTWARE PRIOR TO USE.

4.1 e

Mergency

M

ains

d

isconnection

In case of emergency, the disconnecting point of the equipment is the mains power cord connected to the external power supply, or the mains socket switch.

WARNING: SWITCH OFF THE POWER AT THE MAINS SOCKET, AND REMOVE THE MAINS LEAD FROM THE

EXTERNAL POWER SUPPLY.

4.2 p

ower

-

Up

s

eqUence

Once all system components are connected, it may be turned on as follows:

1. Turn on and log in to the control PC workstation (refer to the supplied manual).

2. Turn on the ILE .

Note switch is located on back panel of ILE

Key switch must be in the ON position for laser operation

3. Turn on the Borealis Conditioning Unit (BCU) .

Ensure Borealis BCU is switched on (position I)

4. Turn on the system camera(s) (refer to the supplied manual).

5. Turn on the filter wheel controller (refer to the supplied manual).

6. Turn on the microscope and associated external devices (refer to the supplied manuals).

7. Turn on the Dragonfly 500 .

On/Off switch located on left side panel

Power button- status illuminates red in standby and green when Dragonfly is active

8. Start up the control software and begin using your system (refer to your software guide e.g. Fusion).


Dragonfly 500

OPERATION

4.3 p

ower

-d

own

s

eqUence

In order to turn the system off, proceed as follows:

1. Exit the control software .

NOTE: This will send all of the device’s motorised stages to their parked positions, allow time for all stages to complete their motion before proceeding to step 2, this will be clearly audible.

2. Turn off the Dragonfly 500 by switching the rocker switch on the left side panel to OFF.

3. Turn off the ILE .

4. Turn off the Borealis Conditioning Unit (BCU) .

5. Turn off the system camera(s) .

6. Turn off the filter wheel controller.

7. Turn off the microscope and associated external devices.

8. Power off the control PC workstation.

4.4 r

isk

M

itigation

4.4.1 M

echanical

h

oUsings

Once installed, the Dragonfly 500 platform, microscope, filter wheel(s), and camera(s) form the protective housings of the product. No components, panels, connections or linkages should be loosened or removed to avoid exposure to hazardous radiation.

4.4.2 h

azards

d

Ue to

M

oistUre or

l

iqUids

Do not put any components including power cables or external power supply in places with high moisture or near water.

In the event of spillage onto the device, immediately power off the device at the mains then try to absorb as much fluid as possible from the exterior of the device. Do not open the device or place your hands into any of the apertures. Call

Andor Customer Support for guidance.


Dragonfly 500

OPERATION

4.5 U

sing the

d

ragonfly

500

Additional technical notes and useful information to help you get the best from your Dragonfly 500 system will be made available from MyAndor , please check the Technical Notes section in case there is further relevant information that is not covered in the scope of this manual

4.5.1 s

tage

c

oVer

For stage cover replacement information please refer to the Dragonfly Technical Notes section of MyAndor .

4.5.2 c

ontrolling the

d

ragonfly

500

throUgh the

s

oftware

Control of the Dragonfly 500 and other integrated system components (ILE laser combiner, microscope, motorized stages, etc.) is handled by the image acquisition control software running on the PC workstation. The software is used to define imaging channels, switch between different imaging modes, and orchestrate/coordinate complex multidimensional acquisition protocols.

The Dragonfly 500 may be controlled through Andor Fusion or Andor iQ image acquisition software packages. An SDK is available on request. Please contact your Andor representative for further information. Refer to your Software Guide for more details on how to setup and operate the Dragonfly 500.


Figure 21: Andor Fusion Software - Specifically Developed for Dragonfly

34

Dragonfly 500

OPERATION

4.6 a

dditional

p

rocedUres

The following sections outline how to exchange emission filter wheels, emission filters and emission splitting optics.

• Access to these optics are provided through the front access panel of the Dragonfly 500 unit.

• The Emission Pathway panel is interlocked to the laser source (ILE) such that if the panel is opened while the

ILE is on, the flow of laser light into the Dragonfly 500 unit will be cut off (see Section 2.5.3

).

• The optics can be exchanged while the ILE is still powered on; however, it is good laser safety practice to power down the ILE during the exchange process.

• Take steps to avoid contamination of the optics with dust i.e. ensure the surrounding area is free of dust, and only open access panels for as long as necessary to perform the required procedure.

• Refer to the general safety information at the start of this manual and the additional procedure specific warnings highlighted for each procedure.

4.6.1 r

eMoVal of the

e

Mission

f

ilter

w

heel

The following procedure describes how to gain access to and remove the emission filter wheels from the Dragonfly 500 unit.

WARNING: When either filter wheel is removed there are two potential hand crush hazards under the following scenarios.

1. If the access door is removed (this requires a tool) the port selection dichroic slider circular aperture hazard

(marked in Yellow below) could be accessible through the T-shaped chamber after the filter wheel was removed. There are hazardous mechanisms below, for the vertical filter wheel, or to the right for the horizontal filter wheel, that could move and hurt fingers if they were placed in this port selection dichroic slider circular aperture.

Moving mechanisms below the circular aperture could move and hurt fingers if placed in the aperture and port selection mechanism activated


In the event of an accident due to this potential hazard:

In software, move the Port Selector to Positions 1 or 2.

35

Dragonfly 500

OPERATION

2. The Magnification Slider Aperture Hazard (marked in blue below) could be accessible through the T-shaped chamber after either filter wheel was removed. There are hazardous mechanisms - above for the vertical filter wheel, or to the left for the horizontal filter wheel, that could move and hurt fingers if they were placed in this circular aperture.

Moving mechanisms through the circular aperture could move and hurt fingers if placed in the aperture and magnification mechanism activated

In the event of an accident due to this potential hazard:

In software, move the Magnification to the previous position (which had allowed access for the finger).

Removing the Filter Wheel

1. If running, quit and exit the image acquisition software.

2. Power off the ILE.

3. Power off the motorized filter wheel controller.

4. Pull on the top-left corner of the front access panel cover to reveal the emission pathway access panel.

5. Unlock the emission pathway panel by unscrewing the three indicated locking screws. Use the provided screwdriver tool stored on the bottom-left corner of the access panel.

Remove using the supplied screwdriver tool

Emission filter wheels are now accessible


Dragonfly 500

OPERATION

6. Swing open the unlocked access panel to reveal the side camera port and top camera port emission filter wheels.

Grab here

Slide Filter

Wheel out

Grab here

7. Pull and disconnect the round 7-pin Fischer connector for the filter wheel that is to be exchanged.

8. Grab and pull out the filter wheel by its sides, as shown above, slide the wheel out fully from its emission pathway slot. Carefully place the filter wheel onto a soft working surface in an upright position to prevent dust accumulating onto the exposed filters.

WARNING: REFER TO THE HAND CRUSH HAZARD WARNINGS OUTLINED AT THE START OF THIS SECTION.

9. Immediately close and lock the front access panel to prevent contamination from dust and airborne particulates.

10. Insert, remove, or replace any desired emission filters in the filter wheel, or prepare a second alternative loaded filter wheel to be put into the Dragonfly 500 unit. Refer to Section 4.6.3 for filter changing procedure.

11. Follow the steps in Section 4.6.2

to re-insert the filter wheel or insert a second alternative filter wheel.


Dragonfly 500

OPERATION

4.6.2 i

nsertion of the

e

Mission

f

ilter

w

heel

The procedure below describes the steps to follow to replace an emission filter wheel back into the Dragonfly 500 unit.

1. Follow Section 4.6.1 to gain access to the filter wheel.

WARNING: REFER TO THE HAND CRUSH HAZARD WARNINGS OUTLINED AT THE START OF SECTION 4.6.2.

2. Holding filter wheel by its sides, fully slide the wheel into its emission pathway slot. Push the wheel all the way into the slot until it hits the physical stop set screw located at the end of the slot.

3. Connect the round 7-pin Fischer connector to its corresponding receptacle. Line up the orange dots on the connector and its receptacle (see image below), and push the connector in until it clicks securely in place.

Alignment mark

4. Immediately close and lock (three screws) the front access panel to prevent and minimize dust and air particulate contamination.

5. Close the square front access panel cover. Push the cover in until it snaps into the fully closed position.

6. If the filter wheel that has been inserted into the Dragonfly 500 is a physically different filter wheel from the one that was previously loaded into the system. The RFID Tag will read the new filter wheel’s details the next time the software is started. It is necessary to adjust the filter wheel offset position that is saved in the filter wheel controller flash memory. If the filters contained in a Filter wheel have been changed then it will be necessary to update the RFID tag with the correct filter details.

7. Power on the FW-1000 motorized filter wheel controller.

8. Power on the lLE.

9. Run the image acquisition control software. Update the filter wheel position names/labels and any image channel definitions as required.


Dragonfly 500

OPERATION

4.6.3 r

eplaceMent of

e

Mission

f

ilters in the

f

ilter

w

heel

For information on emission filter replacement please contact your Andor customer support representative as the emission filter replacement procedure must only be performed by Andor service personnel.


Dragonfly 500

OPERATION

4.6.4 e

xchanging

e

Mission

s

plitting

d

ichroics

The procedure below describes the steps to follow to change any of the Dragonfly 500 emission splitting optics.

WARNINGS:

• Use only emission splitting optics supplied and recommended by Andor.

• There is a hand crush hazard if the power is not turned off before the panel is opened. If the Port

Selection Mirror holder (PSM) is moved from the typical access position to the forward position, fingers placed in the position shown below could be crushed by the moving the PSM against the aperture lip.

In the event of an accident occurring due to this potential hazard, use either one of the following methods:

1. Turn-off Dragonfly and this allows physical pressure on the Holder to open the gap

2. In software, move the Port Selector to Positions 1 or 2

Exchange Procedure

1. In the image acquisition control software, change the emission splitting optic to the Bypass position (position 4).

2. Quit and exit the image acquisition control software.

3. Power off the ILE and Dragonfly 500.

4. Follow Section 4.6.2

to gain access to the filter wheel and emission splitting optics.

5. Open the emission splitting optics access door (Port Selection Mirror Access) using the handle (see below).


Dragonfly 500

OPERATION

6. Use a 3 mm hex driver to unlock the emission splitting optics dovetail slider (~2 turns to loosen the wedge lock screw). Do not remove the screw- only loosen it enough so that the slider may be removed.

Lock screw

Hold handle to remove the emission splitter dichroics

(marked in green)

Emission splitter dichroics

7. Put on a pair of powder-free latex or nitrile gloves. Carefully pull out the dovetail slider and put it down onto a clean, well-lit surface.

8. The Emission Splitter Dichroics may then be exchanged/replaced.

9. Pick up the dovetail slider and push it back into the dovetail slot of the emission optics pathway. Apply forward pressure until the dovetail slider hits the hard stop.

10. While continuing to apply forward pressure, use a 3 mm hex driver to lock the dovetail slider back into place.

To lock the wedge only requires ~2 turns of the wedge lock screw, stop at each 1/2 turn to check if the slider cannot be withdrawn. Once it can’t be withdrawn tighten a further 1/2 turn. Do not overtighten this screw.

11. Close the emission splitting optics access door (Port Selection Mirror Access).

12. Close and lock the Emission Pathway panel.

13. Close the square front access panel cover. Push the cover in until it snaps into the fully closed position.

14. Power on the ILE and Dragonfly 500.

15. Run the image acquisition software. Update the filter wheel position names/labels and any image channel definitions as required.

16. For camera XY alignment adjustment, refer to section 4.6.5.


Dragonfly 500

OPERATION

4.6.5 c

aMera

xy a

djUstMent

Camera xy alignment adjustment should not be necessary under normal use as this is set by Andor service personnel during installation, or if a camera is changed (changing a camera is a service personnel only action). However, if the camera(s) xy position does need re-aligned, for example, if a different primary dual imaging channel has been configured this may be performed as follows:

Notes

• Only adjust the screws indicated below- DO NOT tamper with any other screws.

• Once set during initial installation, the horizontal camera should never need adjustment. The vertical camera camera should be adjusted to account for any relative offset to the horizontal camera. If re-centering is required, this can then be done using the XY stage.

• Use a suitable dual-colour sample, centred in the eye pieces and view in Live Preview mode to facilitate alignment.

• Adjustment screws require 2, 2.5 and 3 mm hex keys.

The basic steps for alignment are as follows:

1. Coarse rotate vertical camera.

2. Coarse translate vertical camera.

3. Fine rotate vertical camera

4. Fine translate vertical camera

The procedure can be divided into several steps:

Focus top camera to confocal disk pinholes

1. Ensure that the top camera is focused to the confocal disk pinholes when mounted to the top camera port

Create and link dual-colour image channels in Fusion

1. In the Fusion software, create two new image channels that use the lasers and emission filters that are compatible with the image splitting dichroic mirror that has been designated for inter-camera registration, as well as the camera lens magnification intended for inter-camera registration. Associate side and top cameras with the corresponding channels that match the direction of emission light transmitted/reflected by the image splitting mirror (In general, shorter wavelengths are reflected with the image splitting dichroic mirrors).

2. Normally two cameras, if fitted, will be the same model, however, if the camera types are not the same (EMCCD and sCMOS), set 2x2 binning for the sCMOS camera.

3. Link the two new image channels such that the images for both channels automatically overlay each other when an image snapped or a live preview is initiated.


Dragonfly 500

OPERATION

Focus both cameras onto multi-colour fluorescent microspheres

1. Engage the primary objective lens (POM) into the optical pathway.

2. Place a multi-colour fluorescent microsphere calibration slide (Life Technologies, product code: T14792) on the microscope stage and focus on the slide coverslip with the POM. Use immersion media if required.

Choose a region of the calibration slide containing microsphere sizes appropriate for the resolution afforded by the microscope objective. For example, use the 200 nm bead size for a high NA oil-immersion 60x or 100x objective lens.

3. Start a 100 ms live preview using the linked dual-colour image channel. Locate a FOV containing as many microspheres as possible, preferably with some microspheres in all four corners of the image.

4. Adjust laser powers and EM gain levels (if present) so that the fluorescent microspheres appear in both channels with equal intensities that fill at least half of the cameras’ dynamic range. Check that spectral crosstalk and bleed-through levels are minimal by turning off each channel laser individually. If significant cross-talk or bleed-through is observed, try re-balancing the channel laser and EM gain levels, or use emission filters that lead to better spectral separation of the fluorescent microspheres.

Rotate and translate top camera

At this point in the procedure, a rotational and lateral offset will likely exist between the two cameras:

A translational adjustment is required

Red and green microspheres overlapped

- perfect registration

1. Insert a 3 mm hex key driver into the top camera port rotational lock access hole and loosen the locking setscrew.

Locking setscrew- ensure the correct screws are identified for adjustment as indicated.

2. With a 2 mm hex key driver turn the fine rotation adjustment setscrew slowly in a clockwise fashion and observe the rotation of the short wavelength component of the live preview image of the fluorescent microspheres. Keep turning the driver until the “clocking-angle” between the corresponding fluorescent microspheres is the same throughout the image FOV.

NOTE: Adjustment of the fine rotation setscrew is unidirectional. If the rotational adjustment of the short wavelength component of the dual-channel image offshoots in the clockwise direction, then the rotational adjustment setscrew must be backed off and the entire camera port mount must be rotated back by hand such that the collar ring tab contacts the setscrew again.

3. Re-tighten the top camera port rotation locking setscrew.


Dragonfly 500

OPERATION

4. Fine-tune the XY adjustment as outlined below.

The X-Y adjustment screws are located as shown in the above images. The adjusters work in opposition to each other e.g. loosen one side, and tighten the other. Note the orientation of the adjustment screws to confirm the axis you are adjusting.

Before starting, lift the plastic port cover to expose the camera port adjustment screws. The cover is held in place with clips. Once loose, it should be lifted and supported to allow easy access to the screws. It could be supported by something, or rotated 45 degrees to rest on the square mount underneath.”

Tighten this side

For the x and y adjustment, there are

2 screws (2 mm) at one side and an opposing screw (2.5 mm) at the other side.


Loosen this side

By loosening one side, and then tightening at the other side, the mount can be translated through the axis of adjustment.

44

Dragonfly 500

MAINTENANCE

Adjusting in the X Axis

Use the adjustment screws shown to translate the image in the x axis. The screws work in opposition to each other, so one side needs to be slackened first, and then the other side may be tightened to move the position. This should be done in increments while checking adjustments in Live Preview mode. Screws at both sides should then be tightened lightly to lock in place, once in the required position.

Adjusting in the Y Axis

Use the adjustment screws shown to translate the image in the y axis. The screws work in opposition to each other, so one side needs to be slackened first, and then the other side may be tightened to move the position. This should be done in increments while checking adjustments in Live Preview mode. Screws at both sides should be tightened lightly to lock in place, once in the required position.


Dragonfly 500

TROUBLESHOOTING

SECTION 5: MAINTENANCE

WARNINGS:

• HERE ARE NO USER MAINTENANCE PROCEDURES REQUIRED FOR THE DRAGONFLY 500 OTHER

THAN THE CLEANING AND CHECKS OUTLINED BELOW.

• THE SYSTEM SHOULD BE POWERED-DOWN PRIOR TO USER PERFORMING ANY MAINTENANCE

CHECKS.

• DO NOT USE EQUIPMENT THAT IS DAMAGED.

• CONTACT YOUR ANDOR REPRESENTATIVE IF THERE ARE ANY QUERIES OR ISSUES WITH YOUR

DRAGONFLY SYSTEM.

5.1 c

leaning and

d

econtaMination

• The most critical aspect of maintenance by the user is to ensure that the system is in a clean environment that is suitable for sensitive electro-optical equipment. The laboratory should be free of dust, fumes and other materials that could affect the system.

• To clean the product, only use a damp, lint-free cloth on the external housing of the unit. Do not wet the connectors.

• Use water only- do not use solvents, cleaning agents, or aerosols.

5.2 r

egUlar

c

hecks

The state of the product should be checked regularly, especially the integrity of the enclosure and the mains cable.

On a Daily Basis :

• Visually inspect the system.

• Perform any maintenance activities suggested by the microscope and camera manufacturer(s).

On a Weekly Basis :

• Ensure that all power cables are firmly in place.

• Check the optical cables and connections to ensure that the locks are in place and no damage has occurred to the optical fibres connecting the various elements of the system.

5.3 a

nnUal

e

lectrical

s

afety

c

hecks

It is advisable to check the integrity of the insulation and protective earth of the product on an annual basis, e.g. U.K.

PAT testing.

5.4 e

xtended

w

arranty

Andor offer extended warranty options for the Dragonfly 500 - please contact your local Andor representative for further information.


Dragonfly 500

TROUBLESHOOTING

SECTION 6: TROUBLESHOOTING

6.1 t

roUbleshooting

e

xaMples

The following examples provide guidance for some basic troubleshooting scenarios. Refer also to the operation and troubleshooting information provided with your other system components. Contact your Andor customer support representative if you require further assistance.

Fault

The BCU is emitting an audible sound.

Image acquisition control software reports a missing system component connection

No laser light observed exiting from the microscope objective lens, sample not illuminated with laser light

Cause

The square fibre is purposefully vibrated for uniform widefield/confocal illumination.

System component not powered on or properly connected to the PC workstation.

ILE not powered on.

ILE not connected to acquisition/ control software.

Microscope side port not activated.

The microscope fluorescence filter turret has one of its filter cubes engaged into the optical pathway that is not compatible with the laser light input from the Dragonfly into the microscope left side port.

The microscope autofocus device uses a dichroic mirror that reflects the ILE laser light instead of transmitting the laser light towards the sample.

Illumination laser wavelength not compatible with the excitation dichroic optic or compatible excitation dichroic optic not present in Dragonfly unit.

An Interlock on the system is not in a safe position (indicated by the two red

“Emission” LEDs on the front of the ILE not illuminating).

The camera(s) are not Live or operating a Protocol and “Active Blanking” is enabled. This synchronises the laser emission with the operation of the camera.

Action

Take no action. A certain level of sound is expected when the BCU is powered on. If the sound seems excessive, contact your nearest Andor

Technical Support Representative.

Quit and exit out of the image acquisition software. Ensure all system components and associated controllers are powered on. Check all cable connections to the PC workstation. Restart the image acquisition control software. If the error persists upon restarting the software, contact your nearest Andor Technical Support Representative.

Turn on ILE mains and turn the key switch to the “on” position. Wait a few minutes for lasers to warm and then restart the image acquisition control software.

Check cabling connections between ILE and computer. Check COM port settings and configure image acquisition control software to communicate with and control ILE.

Ensure microscope is powered on and direct the image to the left side port to which the Dragonfly is connected. Make sure the microscope port selection is an active component of the image channel in the image acquisition control software.

Remove the incompatible filter cube from the optical pathway of the microscope and ensure that any image acquisition channels of the image acquisition control software are configured such that this filter cube is not selected as a component of the channel. An empty fluorescence filter turret is recommended for image channels unless required otherwise for any special applications such as FRAP or photoactivation through another external device.

Contact your nearest Andor Technical Support Representative to determine if the autofocus module dichroic mirror and LED can be replaced with a compatible set of optics that transmits all ILE laser wavelengths.

Choose an excitation dichroic optic that transmits the illumination laser wavelength. If there are no compatible excitation dichroic optics present in the system, contact your local Andor Sales or Support Team representative.

Check that all devices are in a safe operating position.

Go Live in the software or start a Protocol.


Dragonfly 500

TROUBLESHOOTING

Fault

Sample is illuminated with laser light, but little to no fluorescence emission light is detected on the camera, image signal is weak and noisy or blank/absent

Cause

The expression level of the fluorescent probe/contrast agent/stain is too low or did not label the structure of interest within the sample properly. The sample may be bleached already from previous imaging with all fluorophores depleted.

An empty FOV of the sample is being observed.

Action

Consult the fluorescent probe’s manufacturer’s protocol for sample labelling troubleshooting tips or contact the manufacturer directly for additional advice regarding optimal staining. Use a positive control sample with the same fluorescent probe and labelling protocol to verify and validate the experiment.

The sample is not in focus.

Camera is not powered on or image acquisition control software is not properly configured to control the camera.

The wrong camera is being viewed in software.

The wrong imaging channel is being used to view the sample.

The excitation laser wavelength or the emission filter is not appropriate for the fluorescent probe/contrast agent/stain being viewed.

Laterally scan the sample with the microscope stage to find a part of the sample that is fluorescent or use the microscope’s transmitted light imaging modalities (brightfield, DIC, etc.) to locate the specimen of interest.

Switch to a transmitted light imaging mode (brightfield, DIC, etc.) and use the microscope eyepieces to view the sample. Locating the coverslip surface is often easier and faster in this manner. Make sure the microscope is properly set for Koehler illumination with maximum contrast viewing with your eyes. Once the sample coverslip is in focus on your eyes, switch back to viewing the sample through the Dragonfly via one of the fluorescence imaging modes (widefield/confocal/TIRF) and view the sample in camera live preview stream through the image acquisition software. Adjust focus until a strong signal or required plane of focus is achieved.

Turn on the camera. Restart the image acquisition control software and ensure the program properly recognizes the connected camera. If no connection is established, follow the control software instructions to configure the system with the camera. This includes proper installation of any camera driver files onto the system computer (refer to camera manual for driver installation procedures).

Check Fusion for warning messages.

Camera may have failed or not reached operating temperature

The wrong image splitting optic has been selected.

Check that the fluorescence emission light is being properly directed towards the desired camera. Use the correct image splitting optic (100% window, 100% mirror, or dichroic mirror) and update the image acquisition channel settings in the image acquisition software if necessary.

In the image acquisition software, check that the correct camera is active for the desired image channel and update channel settings if necessary.

Ensure that the correct imaging channel is being used to observe the fluorescent contrast agent / probe of interest. Consult the fluorescent probe’s manufacturer’s specification sheet to verify that the imaging channel matches the excitation/emission spectrum of the probe.

The camera exposure time is too low or the EM gain is not activated (EMCCD cameras only).

The ILE laser power is too low.

The camera temperature is too high.

Choose a laser excitation wavelength that lies close to the peak absorption wavelength of the fluorescent probe/contrast agent/stain absorption spectrum being viewed. Choose an emission filter that collects as wide a band of light wavelengths that overlaps as much of the fluorescent probe/ contrast agent/stain emission spectrum without overlapping any of the other laser wavelengths in use in the system. Consult the fluorescent probe manufacturer to find out more about the spectral characteristics of the probe.

Holding the excitation laser power fixed, try increasing the camera exposure time 2-fold, 5-fold, or 10-fold until the image signal strength approaches an acceptable level below the saturation threshold of the camera pixels. If using an EMCCD camera, activate the electron multiplication gain and slowly increase its value until the image signal level is amplified to and acceptable level.

Holding the camera exposure time and gain settings fixed, try increasing the excitation laser power until the image signal strength approaches and acceptable level below the saturation threshold of the camera pixels.

If the sample begins to rapidly photobleach or the signal level does not significantly improve by the time maximum laser power is reached, try increasing the camera exposure time as well.

Ensure that the camera temperature is set to the recommended value and that the room temperature is not above recommended limits. Check the camera manual for further details.

Enable ‘Auto Mapping’ to assist finding signal.

Image intensity mapping may be incorrect or out of range.

The ILE laser light coupling into the optical fibre is low.

Contact your local Andor Sales or Technical Support team.


Dragonfly 500

TROUBLESHOOTING

Fault

Sample is illuminated with laser light, camera pixel intensity is saturated

The system point-spread function (PSF) is blurred or laterally asymmetric

Cause Action

Laser illumination power is set too high. Reduce the laser power in the image acquisition software until the image signal strength is uniformly below the pixel intensity saturation threshold limit.

The wrong image splitting optic has been selected.

The wrong camera is being viewed in software.

The wrong emission filter has been selected or no emission filter is present.

The excitation laser wavelength overlaps with the transmission band of the emission filter.

Check that the fluorescence emission light is being properly directed towards the desired camera. Use the image splitting selection optic (100% window, 100% mirror, or dichroic mirror) and update the image acquisition channel settings in the image acquisition control software if necessary.

In the image acquisition software, check that the correct camera is active for the desired image channel and update channel settings if necessary.

Choose an emission filter that collects as wide a band of light wavelengths that overlaps as much of the fluorescent probe/contrast agent/stain emission spectrum without overlapping any of the other laser wavelengths in use in the system. Consult the fluorescent probe manufacturer to find out more about the spectral characteristics of the probe.

If possible, choose a different laser wavelength that does not overlap with the transmission band of the emission filter, or choose a different emission filter with an emission band or cut-on wavelength that is red-shifted away from the laser wavelength.

Check and reduce the values of these settings if required.

EM Gain set too high or Exposure time is set too long

Transmitted light is contaminating imaging

Widefield illumination selected instead of Confocal.

Turn off room lights or cover the sample with a light weight opaque cover

Switch to confocal mode in the software.

DIC polarization optics are inserted into the optical path.

Remove any DIC polarization optics (i.e: Wollaston prisms) from the objective nosepiece. When imaging the sample via one of the three

Dragonfly fluorescence imaging modes (widefield/confocal/TIRF), it is necessary to clear the optical path of any polarization optics in the microscope.

Ensure that the Cylindrical lens (if present) is not in the imaging path.

Contact your local Andor Sales or Technical Support team representative.

Aberrations (chromatic, comatic, astigmatic, distortion, etc.) are present in the objective lens or microscope optics.

Contaminants or air bubbles are present on the underside of the sample coverslip or the immersion medium.

Clean off the bottom of the coverslip with solvent (methanol or ethanol).

Wipe immersion media from the objective (if present) and clean the front lens of objective lens using standard procedures recommended by the microscope manufacturer. Try re-focusing on the sample using fresh immersion media.

Microscope objective lens may be dirty. Check and clean the microscope objective lens if required.

Image may be saturating Check and adjust exposure and other settings if required.

Camera may have been moved outof-focus with the confocal disk or the

Dragonfly may have been moved outof-focus with the microscope



Dragonfly 500

TROUBLESHOOTING

Fault

The system pointspread function is axially asymmetric

Cause

Sample mounting medium / objective immersion medium refractive index mis-match.

Sample coverslip is the incorrect thickness or is not present.

Objective lens correction collar has not been properly adjusted.

Action

Try to minimize refractive index mis-match between the sample and the objective immersion medium. For example, if using a water immersion objective, try to mount the sample in water. An exception to this rule are

TIRF objective lenses when used for TIRF imaging. For these objectives, the sample must be embedded in water and the objective must use immersion oil.

Always use a coverslip to seal the specimen if the objective lens is designed to image samples through a glass coverslip. Most objective lenses are designed to operate with a #1.5 coverslip thickness (170 m m).

If the objective lens possesses a correction collar, adjust the correction collar dial on the objective while observing the symmetry of a point source of light in the sample as the axial focus varied. Find the collar dial position that minimizes the asymmetry in the axial profile.


The image has a “speckled” appearance that slowly changes with time.

Objective lens has spherical aberrations.

3D Astigmatic Lens is enabled in the software. This optic deliberately adds axial asymmetry to the PSF to allow axial localisation.

BCU not turned on.

Disable the 3D Astigmatic Lens in the software.

Turn on the BCU.

Fibre may be damaged.

Inspect for kinks or breakages

The image intensity rolls off to lower values towards the image edges, the image edges and corners are darker/dimmer/less intense than the central part of the image

The Dragonfly system has been installed with the incorrect mating optics.

The Dragonfly system has been installed with the incorrect mating optics.

The TIRF illumination beam is not collimated as it emerges from the objective lens

TIRF image signals are weaker than expected

The system is using the correct mating optics, but the axial focus of the singlemode optical fibre for TIRF illumination has not been set properly.

Confocal disk inserted into optical pathway.


Illumination uniformity with minimum roll-off or vignetting is achieved only when the correct mating optics are selected for a given microscope setup.

The characteristics of the mating optics are dependent on the presence of additional devices/modules in the microscope infinity space (autofocus modules, extra filter turret decks, objective piezo focus drives, etc.).


A properly collimated TIRF illumination beam can only be achieved when the correct mating optics are selected for a given microscope setup. The characteristics of the mating optics are dependent on the presence of additional devices/modules in the microscope infinity space (autofocus modules, extra filter turret decks, objective piezo focus drives, etc.).


Remove the confocal disk from the optical pathway and if necessary update the image channel settings in the image acquisition software.

Penetration depth set incorrectly. Not at critical angle.

Check and adjust the settings if required.


Dragonfly 500

TROUBLESHOOTING

Fault Cause

The illumination size (power density) is set to a size that does not match the full camera FOV.

Action

In the image acquisition software, choose the largest illumination size

(usually power density setting 1)

The widefield/confocal image is unexpectedly only visible in a sub-region of the full camera FOV

The Dragonfly field diaphragm is closed down to a size that is too small for the camera.

In the image acquisition software, choose the largest field diaphragm size that best matches the full camera FOV.

Wrong Excitation Aperture has been selected. This would mask part of the illumination).

Check and update the settings if required.

BCU square multi-mode fibre adapter has physically rotated in its mount.

The widefield/confocal illumination is not square to the camera

One or both cameras have been physically rotated or not properly connected to their respective camera port mount.

Image unexpectedly exhibits a magnification not equal to the magnification of the objective lens


For laser safety reasons, only qualified Andor personnel should correct a fibre adapter rotation error like this

See Section 2.4 of this document which describes the required camera orientations when connected to the Dragonfly camera ports.

The Dragonfly passes the microscope image to the camera with no additional magnification only when the 1x camera zoom lens is selected.

In the image acquisition software, ensure that the 1x camera zoom lens is selected.

Image unexpectedly exhibits a magnification not equal to the magnification of the objective lens

The image acquisition software is set to view a cropped ROI of the full camera field of view or the image is being viewed with a digital zoom factor greater than 100%.

Values for objectives set in software are incorrect.

In the image acquisition software, set the camera output to full frame (no crop ROIs) or set the digital zoom factor for the image to 100%.

Check and update values in the software if required.

Images acquired simultaneously on two identical cameras are no longer in registry with each other.

Images acquired simultaneously on two identical cameras rotated/ mirrored with respect to each other.

One of the camera ports has been moved out of alignment

Cameras have different magnification values.


Ensure both cameras are using the same magnification setting.

Optimal registration is only achievable for one combination of excitation dichroic, image splitting dichroic and camera magnification.

Image acquisition software not properly configured for two-camera imaging.

One or both cameras have been physically rotated or not properly connected to their respective camera port mount.

Use the image acquisition software’s image rotation and mirroring tools to make the image orientation of one camera to match the orientation of the image presented by the other camera. Ideally, the images of both cameras are oriented to match the orientation of the image as seen through the microscope eyepieces.

See Section 2.4

of this document which describes the required camera orientations when connected to the Dragonfly camera ports.


Dragonfly 500

TECHNICAL SPECIFICATIONS

6.1.1 p

robleM

r

eporting

f

orM

Please have the following information when connecting customer support:

Owner Information

Institution Name

Vendor

Model

Vendor Information

Contact

Equipment Information

Serial No.

Check List

Cable Connections

Mains Power

What is the nature of the problem?

Summary

Any other observations?


Dragonfly 500

MECHANICAL DRAWINGS

SECTION 7: TECHNICAL SPECIFICATIONS

7.1 g

eneral

s

pecifications

Confocal/Widefield Input Wavelength

Range

TIRF Input Wavelength Range

Confocal/Widefield Input Power

Output Power

Confocal Pinhole Diameter

Lateral resolution

Maximum Frame Rate - Confocal (fps)

Switching time - Confocal/TIRF

Weight

Dimensions cm [inches]

Minimum support table dimensions

Excitation range: 400-800 nm

Emission range: 420-850 nm

400-640 nm

2W maximum for combined wavelengths

As classified under IEC 60825-1 or the regional equivalent: 500 mW maximum Class 3B

(typical). May be Class 4 with 1W maximum with some laser combinations – refer to Laser

Safety and Classification sections in the preface of this manual.

25 m m and 40 m m

Diffraction limited in 19 mm diagonal

400

4 seconds

60 kg approx. max without cameras/accessories.

31.2 x 30.7 x 19.3 [792 x 780 x 489] (with installation handles)

31.2 x 26.5 x 19.3 [792 x 674 x 489] (without handles)

122 x 92 cm[48 x 36 inches] approx. (depends on configuration)

7.2 e

nVironMental

s

pecifications

Usage

Altitude

Operating Temperature

Storage Temperature

Indoor use only

Up to 2000 m

18°C to 28 °C ambient

0°C to 50 °C

Operating Relative Humidity

Overvoltage Category

<70% (non-condensing)

CAT II. An overvoltage category of CAT II means that the equipment is designed to cope with transient voltages above the rated supply that would be experienced by any product connected to a mains socket in a building.

Rated Pollution Pollution Degree 2. Normally only non-conductive pollution occurs. Occasionally, however, a temporary conductivity caused by condensation must be expected.

Ventilation Requirements Do not cover during operation.

7.3 p

ower

s

Upply

r

eqUireMents

Mains Input

Low Voltage Output

Low Voltage Output Ripple

100-240 V a.c. 50-60 Hz

24 V d.c. ± 2%

480 mV max.

Low Voltage Steady State Current Output 5 A max.*

Low Voltage In-rush Current Capability Shall start up a load whose in-rush current from a 0.1 Ω source resistance is 3A min. peak and a pulse width of 20 ms min. measured at half the peak

Low Voltage Power Supply Output

Connector

Fischer S104 A037-230+ (shielded cable connector) which uses UL 94 V-0 PEEK insulators

E3 104.3/7.7 + B (clamp set)

Pins 1,2 = +24V

Pins 3,4 = 0V Return

General Safety Requirements The power supply must be certified to an appropriate IEC standard, e.g. IEC 60950-1, and meet the reinforced insulation from mains requirement of IEC 61010-1

Power Consumption Typical/ Maximum: 65 W/ 120 W


Dragonfly 500

GLOSSARY

APPENDIX A: MECHANICAL DRAWINGS

Dimensions in mm[inches]

Note: Shown without installation handles, depth is 780 [30.7] with handles.

For dimensions of the ILE, please refer to the ILE specifications sheet .


Dragonfly 500

OTHER INFORMATION

APPENDIX B: GLOSSARY

AC

ADC

Alternating Current (Mains).

AC/DC An electronic device that converts AC electricity (usually mains) into a DC voltage (usually a safe

Analogue-to-Digital Converter: Converts an analogue voltage to a digital signal.

AEL Accessible Emission Limit: The maximum accessible emission permitted within a particular class.

Wherever the text refers to “emission level not exceeding the AEL” or similar wording, it is implicit that the accessible emission is determined following the measurement criteria specified in Clause 9 of IEC

BCU

60825-1.

Beam Conditioning Unit: Homogenizes the light from the laser source to permit uniform illumination across

CSU

CLSM

DC

Confocal Spinning Disk.

Confocal Laser Scanning Microscopy.

Direct Current.

Diffuse Reflection Change of the spatial distribution of a beam of radiation by scattering in many directions by a surface or

medium.

EMC

ESD

EU

FC/APC

Electromagnetic Compatibility.

Electrostatic Discharge.

European Union.

Fibre Optic connector (Angled Physical Contact).

Input/Output: Generic input and output electrical signal connections.

IO

ILE

Intrabeam Viewing

LED

MM

NOHD

MPE

Parfocal

PC

PD

POL

PSF

Specular Reflection

Integrated Laser Engine: Control for up to 8 individual lasers.

All viewing conditions whereby the eye is exposed to the direct or specularly reflected laser beam in contrast to viewing of, for example, diffuse reflections.

Light Emitting Diode.

Multi-mode fibre: Multi-mode fibres have a much larger core diameter than single mode fibres.

This enables improved transmission of longer wavelengths.

Nominal Ocular Hazard Distance: Distance from the output aperture at which the beam irradiance or radiant exposure equals the appropriate corneal maximum permissible exposure (MPE)

Maximum Permissible Exposure: Level of laser radiation to which, under normal circumstances, persons may be exposed without suffering adverse effects. The MPE levels represent the maximum level to which the eye or skin can be exposed without consequential injury immediately or after a long time and are related to the wavelength of the laser radiation, the pulse duration or exposure duration, the tissue at risk and, for visible and near infra-red laser radiation in the range 400 nm to 1 400 nm, the size of the retinal image. Maximum permissible exposure levels are (in the existing state of knowledge) specified in

Annex A of IEC 60825-1.

When optical components- such as the objective lenses or camera ports of a microscope are described as parfocal this means that they may be interchanged without affecting the focus on the image.

Personal computer.

Dragonfly illumination Power Density level.

Primary Objective Lens: The primary objective lens is defined as the objective lens with which most imaging will be conducted when using the microscope.

Point Spread Function.

Reflection from a surface that can be considered a beam, including reflections from mirrored surfaces.

For further technical information and tutorials please visit the Andor Learning Academy on our website.


Dragonfly 500

OTHER INFORMATION

APPENDIX C: OTHER INFORMATION

T erms and

C ondiTions of

s ale and

W arranTy

i nformaTion

The terms and conditions of sale, including warranty conditions, will have been made available during the ordering process. The current version may be viewed at: www.andor.com/pdfs/literature/Andor_Standard_Warranty.pdf

W asTe

e leCTroniC and

e leCTriCal

e quipmenT

r egulaTions

2006 (Weee)

The company’s statement on the disposal of WEEE can be found in the Terms and Conditions


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