Princeton Instruments MICROMAX SYSTEM Manual de usuario

4411-0039-CE

Version 4.A

November 26, 2001



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Trenton, NJ 08619

TEL: 609-587-9797

FAX: 609-587-1970

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registered trademarks of Microsoft Corporation.

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The information in this publication is believed to be accurate as of the publication release date. However,

Roper Scientific, Inc. does not assume any responsibility for any consequences including any damages

resulting from the use thereof. The information contained herein is subject to change without notice.

Revision of this publication may be issued to incorporate such change.

iii

Table of Contents

Chapter 1 Introduction ........................................................................................ 9

Introduction.........................................................................................................................9

MicroMAX System Components........................................................................................9

Overview ......................................................................................................................9

Camera..........................................................................................................................9

Controller....................................................................................................................10

Applications................................................................................................................11

Computer Requirements....................................................................................................11

About this Manual.............................................................................................................12

Manual Organization ..................................................................................................12

Safety Related Symbols Used in This Manual...........................................................13

Environmental Conditions ................................................................................................13

Grounding and Safety .......................................................................................................13

Precautions........................................................................................................................14

Repairs ..............................................................................................................................14

Cleaning ............................................................................................................................14

Camera and Controller ...............................................................................................14

Optical Surfaces..........................................................................................................14

Roper Scientific Customer Service...................................................................................14

Chapter 2 Installation Overview ....................................................................... 15

Chapter 3 System Setup ................................................................................... 17

Unpacking the System.......................................................................................................17

Checking the Equipment and Parts Inventory...................................................................17

Power Requirements .........................................................................................................18

Verifying Controller Voltage Setting................................................................................18

Mounting the Camera........................................................................................................19

General .......................................................................................................................19

Mounting the Lens......................................................................................................19

Mounting to a Microscope..........................................................................................20

Installing the Application Software ..................................................................................24

Drivers ........................................................................................................................24

Installing the Interface Card..............................................................................................24

Connecting the TAXI (Controller-Computer) Cable........................................................25

Connecting the Detector-Controller Cable or the Camera Power/Camera Signal

Cables................................................................................................................................25

iv MicroMAX System User Manual Version 4.A

Chapter 4 Operation .......................................................................................... 27

Introduction.......................................................................................................................27

EMF and Xenon or Hg Arc Lamps...................................................................................27

Vacuum .............................................................................................................................27

Cooling..............................................................................................................................27

Setting the Temperature .............................................................................................28

Temperature Stabilization ..........................................................................................28

Baseline Signal..................................................................................................................28

Imaging Field of View ......................................................................................................29

RS-170 or CCIR Video .....................................................................................................29

First Light..........................................................................................................................31

Cabling........................................................................................................................32

Getting Started............................................................................................................33

Setting the Parameters ................................................................................................33

Focusing......................................................................................................................34

Acquiring Data ...........................................................................................................36

Summary ...........................................................................................................................36

Chapter 5 Timing Modes ................................................................................... 37

Full Speed or Safe Mode...................................................................................................37

Standard Timing Modes....................................................................................................38

Free Run .....................................................................................................................38

External Sync..............................................................................................................40

External Sync with Continuous Cleans ......................................................................42

Frame Transfer Operation.................................................................................................43

Interline Operation ............................................................................................................45

Operating Modes ........................................................................................................45

Timing Options in Overlapped Readout Mode ..........................................................46

Chapter 6 Exposure and Readout .................................................................... 49

Exposure............................................................................................................................49

Exposure with an Interline Array ...............................................................................50

Exposure with a Mechanical Shutter..........................................................................50

Exposure with an Image Intensifier............................................................................51

Continuous Exposure (no shuttering).........................................................................51

Saturation....................................................................................................................52

Dark Charge................................................................................................................52

Array Readout ...................................................................................................................53

Full Frame...................................................................................................................53

Frame Transfer ...........................................................................................................55

Interline.......................................................................................................................56

Binning .......................................................................................................................59

Digitization........................................................................................................................62

Dual A/D Converters..................................................................................................62

Table of Contents v

Chapter 7 MicroMAX DIF Camera (Double Image Feature)............................ 65

Introduction.......................................................................................................................65

Timing Modes ...................................................................................................................66

Free Run .....................................................................................................................66

IEC (Internal Exposure Control) ................................................................................68

EEC (External Exposure Control)..............................................................................70

ESABI (Electronic Shutter Active Between Images).................................................71

Tips and Tricks .................................................................................................................72

Lab Illumination .........................................................................................................72

Background Subtraction .............................................................................................72

Flatfield Correction ....................................................................................................73

Mask Bleed-Through Correction................................................................................73

Chapter 8 TTL Control....................................................................................... 75

Introduction.......................................................................................................................75

TTL In ...............................................................................................................................75

Buffered vs. Latched Inputs ..............................................................................................76

TTL Out ............................................................................................................................76

TTL Diagnostics Screen ...................................................................................................77

Hardware Interface............................................................................................................77

Example......................................................................................................................78

Chapter 9 System Component Descriptions................................................... 79

MicroMAX Camera ..........................................................................................................79

ST-133 Controller .............................................................................................................82

Cables................................................................................................................................86

Interface Card....................................................................................................................86

Application Software ........................................................................................................87

User Manuals ....................................................................................................................87

Chapter 10 Troubleshooting............................................................................. 89

Introduction.......................................................................................................................89

Baseline Signal Suddenly Changes...................................................................................89

Changing the ST-133's Line Voltage and Fuses ...............................................................89

Controller Is Not Responding ...........................................................................................90

Cooling Troubleshooting ..................................................................................................91

Temperature Lock cannot be Achieved or Maintained. .............................................91

Detector loses Temperature Lock...............................................................................91

Gradual Deterioration of Cooling Capability.............................................................91

Detector Stops Working....................................................................................................92

Error occurs at Computer Powerup...................................................................................92

Removing/Installing a Plug-In Module.............................................................................95

Shutter Malfunctions.........................................................................................................96

Appendix A Specifications ............................................................................... 97

CCD Arrays.......................................................................................................................97

Spectral Range............................................................................................................97

Types ..........................................................................................................................97

vi MicroMAX System User Manual Version 4.A

Temperature Control.........................................................................................................97

Cooling..............................................................................................................................98

Mounting...........................................................................................................................98

Shutters..............................................................................................................................98

Inputs.................................................................................................................................99

Outputs..............................................................................................................................99

Programmable Interface ....................................................................................................99

A/D Converter...................................................................................................................99

Computer Requirements....................................................................................................99

Miscellaneous .................................................................................................................100

Appendix B Outline Drawings ........................................................................ 101

Appendix C Kinetics Mode ............................................................................. 107

Introduction.....................................................................................................................107

Kinetic Timing Modes ....................................................................................................108

Free Run ...................................................................................................................108

Single Trigger...........................................................................................................109

Multiple Trigger .......................................................................................................109

Appendix D Virtual Chip Mode ....................................................................... 111

Introduction.....................................................................................................................111

Virtual Chip Setup ..........................................................................................................112

Introduction ..............................................................................................................112

Equipment:................................................................................................................112

Software:...................................................................................................................112

Assumptions: ............................................................................................................113

System Connection Diagram:...................................................................................113

Procedure:.................................................................................................................113

Experimental Timing ......................................................................................................116

Virtual Chip dialog box ..................................................................................................116

Tips..................................................................................................................................117

Appendix E Repumping the Vacuum............................................................. 119

Introduction.....................................................................................................................119

Requirements ..................................................................................................................119

Vacuum Pumpdown Procedure.......................................................................................120

Warranty & Service.......................................................................................... 123

Limited Warranty: Roper Scientific Analytical Instrumentation....................................123

Limited One (1) Year Warranty ...............................................................................123

Sealed Chamber Integrity Warranty.........................................................................123

Vacuum Integrity Warranty......................................................................................123

Image Intensifier Detector Warranty........................................................................123

X-Ray Detector Warranty.........................................................................................124

Software Warranty....................................................................................................124

Owner's Manual and Troubleshooting......................................................................124

Your Responsibility..................................................................................................124

Contact Information ........................................................................................................125

Table of Contents vii

Index ................................................................................................................. 127

Figures

Figure 1. MicroMAX Cameras and Controller..................................................................9

Figure 2. Standard System Diagram ................................................................................16

Figure 3. 5 MHz System Diagram ...................................................................................16

Figure 4. Controller Power Input Module........................................................................18

Figure 5. Bottom Clamps .................................................................................................22

Figure 6. Bottom Clamp secured to Relay Lens ..............................................................23

Figure 7. WinView Installation: Interface Card Driver Selection ...................................24

Figure 8. Imaging Field of View......................................................................................29

Figure 9. Monitor Display of CCD Image Center Area...................................................30

Figure 10. Standard System Connection Diagram...........................................................31

Figure 11. 5 MHz System Diagram .................................................................................32

Figure 12. F-mount Focus Adjustment ............................................................................36

Figure 13. Chart of Full Speed (Synchronous) and Safe (Asynchronous) Operation.....39

Figure 14. Free Run Timing Chart (part of the chart in Figure 13) .................................40

Figure 15. Free Run Timing Diagram..............................................................................40

Figure 16. Chart Showing Two External Sync Timing Options......................................41

Figure 17. External Sync Timing Diagram ......................................................................41

Figure 18. Continuous Cleans Flowchart.........................................................................42

Figure 19. Continuous Cleans Timing Diagram ..............................................................43

Figure 20. Frame Transfer where tw1 + texp + tc< tR...................................................44

Figure 21. Frame Transfer where tw1 + texp + tc> tR...................................................45

Figure 22. Frame Transfer where Pulse arrives after Readout ........................................45

Figure 23. Overlapped Mode where tw1 + texp + tc< tR...............................................47

Figure 24. Overlapped Mode where tw1 + texp + tc> tR...............................................47

Figure 25. Overlapped Mode where Pulse arrives after Readout ....................................48

Figure 26. Block Diagram of Light Path in System.........................................................49

Figure 27. CCD Exposure with Shutter Compensation ...................................................51

Figure 28. Full Frame at Full Resolution.........................................................................53

Figure 29. Frame Transfer Readout .................................................................................55

Figure 30. Overlapped Mode Exposure and Readout......................................................57

Figure 31. Non-Overlapped Mode Exposure and Readout..............................................58

Figure 32. 2 × 2 Binning for Full Frame CCD.................................................................60

Figure 33. 2 × 2 Binning for Interline CCD.....................................................................61

Figure 34. Free Run Mode Timing Diagram ...................................................................67

Figure 35. Setup using READY to Trigger an Event.......................................................67

Figure 36. Timing for Experiment Setup shown in Figure 35.........................................67

Figure 37. Timing Diagram for Typical IEC Measurement.............................................69

Figure 38. Setup for IEC Experiment with Two Lasers ..................................................69

Figure 39. Timing Diagram for IEC Experiment with Two Lasers.................................69

Figure 40. Another Hardware Setup for an IEC Measurement........................................70

Figure 41. EEC Timing Example with Exposure Time in Software Set to texp................71

Figure 42. ESABI Timing Example: Image Exposure time = texp set in software ...........72

Figure 43. TTL IN/OUT Connector.................................................................................77

Figure 44. Controller Front Panel ....................................................................................82

Figure 45. ST-133 Rear Panel..........................................................................................83

Figure 46. SCAN and SHUTTER MONITOR Signals....................................................86

viii MicroMAX System User Manual Version 4.A

Figure 47. Power Input Module .......................................................................................90

Figure 48. Fuse Holder.....................................................................................................90

Figure 49. Module Installation.........................................................................................95

Figure 50. Controller Dimensions..................................................................................101

Figure 51. Rectangular Camera Head: C-Mount ...........................................................102

Figure 52. Rectangular Camera Head: F-Mount............................................................103

Figure 53. 1 MHz and 100kHz/1MHz Round Head Camera: C-Mount Adapter and

Shutter.....................................................................................................................104

Figure 54. 5 MHz Round Head Camera: C-Mount Adapter..........................................105

Figure 55. 1 MHz Round Head Camera: F-Mount Adapter ..........................................106

Figure 56. Kinetics Readout ..........................................................................................107

Figure 57. Hardware Setup dialog box ..........................................................................108

Figure 58. Experiment Setup dialog box .......................................................................108

Figure 59. Free Run Timing Diagram............................................................................109

Figure 60. Single Trigger Timing Diagram ...................................................................109

Figure 61. Multiple Trigger Timing Diagram................................................................110

Figure 62. Virtual Chip Functional Diagram.................................................................111

Figure 63. System Diagram............................................................................................113

Figure 64. Virtual Chip dialog box. ...............................................................................116

Figure 65. Vacuum Connector Required for Pumping ..................................................120

Figure 66. Removing the Back Panel.............................................................................120

Figure 67. Attaching the Vacuum Connector ................................................................121

Figure 68. Opening the Camera to the Vacuum System................................................121

Tables

Table 1. Bottom Clamps for Different Microscopes .......................................................22

Table 2. Camera Timing Modes ......................................................................................37

Table 3. Approximate Readout Time for the Full-Frame CCD Array.............................54

Table 4. Approximate Readout Time for the Frame-Transfer CCD Array .....................55

Table 5. Approximate Readout Time for the Interline CCD Arrays ...............................59

Table 6. Readout Rates for PI 1300 × 1030 Array at 1 MHz ..........................................59

Table 7. Well Capacity for some CCD Arrays ................................................................62

Table 8. Bit Values with Decimal Equivalents: 1 = High, 0 = Low...............................76

Table 9. TTL IN/OUT Connector Pinout ........................................................................77

Table 10. ST-133 Shutter Drive Selection.......................................................................85

Table 11. I/O Address & Interrupt Assignments before Installing Serial Card...............93

Table 12. I/O Address & Interrupt Assignments after Installing Serial Card..................93

Table 13. MicroMAX:512BFT: Virtual Chip Size, Exposure Time, and Frames per

Second.....................................................................................................................112

Chapter 1

Introduction

The Princeton Instruments®MicroMAX system is a high-speed, low-noise CCD camera

system designed for demanding imaging applications and is an optimal system for use in

fluorescence microscopy applications such as high resolution immunofluorescence, FISH

or GFP imaging. The MicroMAX system incorporates a compact camera head, cooled

CCD, advanced exposure-control timing, video output, and sophisticated readout

capabilities.

Among the advantages of the MicroMAX concept are the range of CCD arrays available

and the built-in video output mode. The system can be configured either with a variety of

interline CCDs to provide true 12-bit images at a readout rate of up to 5 million pixels

per second, or with a number of back-illuminated CCDs to provide true 16-bit images.

The built-in video output mode simplifies setup and focusing on the microscope. The

combination of the MicroMAX system with one of a variety of specialty software

packages results in a powerful digital imaging system that can meet most experimental

needs.

MicroMAX System Components

Overview

The MicroMAX imaging system consists

of a camera (either a round head or a

rectangular head depending on

application), controller, digital interface

card, a computer, cables, manuals, and

application software. Together, these

components allow you to acquire

quantitative digital data under very low

light imaging conditions. Each component

is optimized for its specific function. In

operation, data acquired by the camera is

Figure 1. MicroMAX Cameras and Controller

routed to the controller and from there to the computer for processing and display. A

composite video output allows immediate viewing of the acquired images on a separate

monitor. The application software (for example, Princeton Instruments WinView/32)

allows the computer to control both the system configuration and data acquisition.

Camera

Introduction: The function of the camera is to collect very low intensity light and

convert the energy into a quantitative, electronic signal (photo-electrons) over a two

dimensional space. To do this, light from the subject is focused onto a CCD array, which

10 MicroMAX System User Manual Version 4.A

accumulates photoelectrons for the exposure time. At the end of the exposure time, the

image thus formed is read out. The accumulated charge on each cell of the array is

transferred out of the CCD array, amplified, and sent to the controller as an analog

signal, where it is digitized prior to transfer to the computer.

The camera is highly integrated, containing the shutter (if applicable) and thermoelectric

cooler with optional forced-air supplemental cooling in a single, shielded housing.

Surface mount electronic technology is used wherever possible, giving a compact

package with uncompromising performance.

Depending on your application, the camera included in your MicroMAX system will be

either a compact round camera head or a high performance, cooled, rectangular camera

head. The round head features interline CCDs; its small size ensures that the camera can

be mounted on virtually any microscope port, including those found on inverted

microscopes. The rectangular head features back-illuminated CCDs with frame transfer

readout.

At the heart of the camera is the CCD array centered on the optic axis. Available formats

include the:

➧EEV CCD57-10, 512×512, 13×13µm pixels for the MicroMAX:512BFT

➧EEV CCD47-10, 1024×1024, 13×13µm pixels for the MicroMAX:1024B

➧Sony ICX075, 782×582, 8.3× 8.3µm pixels for the MicroMAX:782Yand the

MicroMAX:782YHS systems

➧Sony ICX061,1300×1030, 6.7× 6.7µm pixels for the MicroMAX:1300Y, the

MicroMAX:1300YHS, and MicroMAX:1300YHS-DIF systems

A special clocking mode to minimize background signal is supported. See the Roper

Scientific brochures and data sheets for detailed specifications.

Cooling System: MicroMAX cameras have a multi-stage Peltier type cooler that is

thermally coupled to the CCD surface. Heat is sequentially transferred through the

Peltier stages and from there to the outer shell of the camera via a heat transfer block.

This cooling system allows the camera to maintain CCD temperature of typically -15C

for round cameras head and -45°C for rectangular camera heads. Cameras equipped with

a fan assembly can reach lower CCD temperatures for reduced thermal noise and

extended exposure times.

Low Noise Readout: In order to achieve a low-noise readout of the CCD, several

design features have been implemented. These include cooling the preamplifier on the

CCD, isolating circuits to prevent electronic crosstalk and minimizing the path lengths of

critical electronic circuits. The net result of these design features is the lowest available

readout noise at the highest speed possible for these CCDs.

Controller

Data Conversion: The controller accepts the analog data and converts it to digital data

using specially designed, low-noise electronics supporting scientific grade 12- or 16-bit

Analog to Digital (A/D) converters.

The standard MicroMAX Controller enables both high-speed and high-precision readout

capabilities. It can collect 16-bit images at a readout rate of up to 1 million pixels per

second (1 MHz) in the high-speed mode or at 100 thousand pixels per second (100 kHz)

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