P/N ICCD. Detector. Operation manual

Transcription

1 P/N ICCD Detector Operation manual Manual Version 3 Revision F July 9, 1999

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3 Table of Contents Chapter 1 General Information... 7 Environmental and Operating Conditions...8 Physical Features...9 Preamplifier Selector switch...10 Chapter 2 Detector Setup General Instructions...13 Connecting the detector...13 Setting the controller...13 Electronic Control of the Image Intensifier...13 Shutter (CW) operation...14 Operation with a PI Pulser and a Gen II Intensifier...14 Operation with a Gen IV Intensifier...15 Operation with the FG Imaging Applications...16 Connecting lenses...17 Overexposure protection...17 Spectroscopic Applications...17 Model IVUV...18 System Connections...19 Chapter 3 Flushing and Cooling Nitrogen Flushing of the Detector...21 Coolant Connections...22 Setting the Temperature...23 CPC-100 Photocathode Cooler Option...24 Connections and operation...24 Retrofit...26 Chapter 4 Focusing System Connections...27 Precautionary Measures...27 Laboratory conditions...27 Condensation...28 Image intensifier gain...28 iii

4 iv ICCD Detector manual Version 3.F Adjustment precautions...28 Alarm...29 Power Up...29 Baseline Signal...29 Shutter vs. Gated Operation...30 Focusing Imaging Systems...31 Imaging field of view...31 Spectroscopy Systems...32 Kinetics ICCD with Moveable Mask...33 Chapter 5 Microscopy Applications Introduction...35 Mounting the Camera on the Microscope...35 F-Mount...36 C-Mount...38 Operation...38 Xenon or Mercury Arc Lamp Precautions...38 Focusing the Microscope...39 Adjusting the Parfocality of the Camera...39 Imaging Hints...39 Fluorescence...40 Microscopes and Infrared Light...40 Chapter 6 FG-101 Pulser Single TTL Input...41 Delay Generator Input Pin Connector Signals...42 Appendix A Problems and Solutions Temperature Lock Problems...45 Detector doesn t achieve temperature lock...45 Detector loses temperature lock...45 Intensifier Alarm...45 Excessive Readout Noise...46 Appendix B ICCD Detector Specifications Common ICCD Specifications...47 Operating parameters...47 Operating environment...48 ICCDs with the CPC-100 Option...48 ICCDs with the FG-101 Option...49 Kinetics ICCDs...50

5 Table of Contents v Appendix C Changing the Internal Impedance of an ICCD Measuring the Impedance of an ICCD Detector...51 Changing from 50 to 1 M...52 Changing from 1 M to ICCD Detectors for the FG-100H...53 Appendix D UV Lens Specifications...56 Installing the Camera in the Stand...56 Installing the Lens...58 Nitrogen Flushing of the Detector...61 Adjustments...61 Elevation...62 Angulation...62 Lateral Adjustments...63 Filter Installation...65 Conversion...69 Removing the UV Lens...69 Removing the Fixed Light Seal...70 Removing the Camera...73 Warranty and Service Warranty...75 Equipment Repairs...75 Contact Information...76 Index Figures Figure 1. The ICCD Detector...7 Figure 2. ICCD detector shown with and without the CPC-100 option...8 Figure 3. Size comparisons for 1:1 coupling...9 Figure 4. View of standard and FG-101 ICCD detectors from the rear...10 Figure 5. ICCD with Thomson chip and Preamplifier Selector switch...11 Figure 6. Nikon lens adapter...16 Figure 7. Deep focal plane adapters...18 Figure 8. System connection diagram...19 Figure 9. Temp. knob, located on the front of the controller...23 Figure 10. Do not bang the connections to the CPC-100!...24 Figure 11. Making connections to the CPC Figure 12. Standard CPC-100 coolant connections...26 Figure 13. Intensifier gain adjustment...28 Figure 14. Imaging field of view...31 Figure 15. ICCD with kinetics option, viewed from below...34

6 vi ICCD Detector manual Version 3.F Figure 16. Diagnostic Instruments F-mount adapters...37 Figure 17. Bottom clamp secured to relay lens...37 Figure pin connector for delay generator...42 Figure high voltage board...53 Figure 20. ICCD Detector with PI f/1.2 UV Lens...55 Figure 21. The Stand...57 Figure 22. Camera Mounted in Stand...58 Figure 23. UV Lens and Mounted ICCD...59 Figure 24. Lens Engaged and in Position to Mate the Floating Light Seal with the Fixed Light Seal...59 Figure 25. UV Lens with Front Shield and Dust Cap...60 Figure 26. Elevation Thumb Nuts...62 Figure 27. Back View of ICCD and Stand...63 Figure 28. Left Side View...64 Figure 29. Right Side View...64 Figure 30. Filter and Filter-Mounting Components...65 Figure 31. Profile Drawing of Filter-Mounting Components...65 Figure 32. UV Lens, Camera, Front Shield and Filter Components...66 Figure 33. UV Lens with First Spacer (only) Installed...67 Figure 34. UV Lens with First Spacer and Filter Installed...67 Figure 35. Second Spacer Installed...68 Figure 36. Retainer Installed...69 Figure 37. Fixed Light Seal Securing Screws...71 Figure 38. Fixed Light Seal Removed; Gas Tubing Still Connected...71 Figure 39. Disconnecting the Gas Tubing...72 Figure 40. Spectroscopy Nose Installed...72 Figure 41. Removing the Camera...73 Figure 42. Conversion Components Grouping...74

7 General Information Chapter 1 Figure 1. The ICCD Detector WARNING Image intensified detectors (ICCDs), Figure 1, can be destroyed if exposed to excessive light levels. Operating ICCD detectors without water or other coolant circulating may also destroy the detector. RS Princeton Instruments (PI) cannot take responsibility for ICCD detector damage due to misuse. Image intensified CCD detectors (ICCDs) are ideal for applications involving ultra low light measurements, or measurements of transient effects. They utilize a proximityfocused microchannel plate (MCP) image intensifier fiber-optically coupled to a CCD array. Standard proximity-focused MCPs have PMT-like UV-NIR response, linear geometric accuracy, and the fastest intensifiers can be gated in as little as 2 nsec with an exceptionally high on/off shutter ratio. The CCD array provides a low noise, high dynamic range readout device that can be scanned at a variety of pixel rates. WARNING The input impedance of the Gate BNC connector could be any one of three different values, each intended for operation with a different type pulser. Operating the ICCD with the Gate BNC connected to the wrong pulser could cause permanent damage to the ICCD detector and the pulser. Damage due to misuse is not covered by warranty. 7

8 8 ICCD Detector manual Version 3.F Environmental and Operating Conditions Storage Temperature: 55 C Lab Temperature: 30 C > T > -25 C Water Flow: 1-2 liters per minute for maximum cooling Nitrogen Gas: 2-3 liters per minute initially, ml per minute during experiment Lab Humidity: < 50%. In very high humidity climates any system optics close to the image intensifier (such as a spectrometer) must also be continuously flushed with dry nitrogen gas to prevent condensation. Figure 2. ICCD detector shown with and without the CPC-100 option Nitrogen Gas Inlet Standard ICCD, viewed from above Coolant Ports CCD Image Intensifier ICCD with CPC-100 option, viewed from the side CCD Coolant Port Clamp Image Intensifier Nitrogen Gas Inlet

9 Chapter 1 General Information 9 Physical Features An ICCD detector has several distinct sections. The front enclosure contains the image intensifier and the CCD. The image intensifier is coupled to the CCD using a fiber optic window on both the output of the image intensifier and the front side of the CCD. For most models of ICCD the fiber optics are 1:1, translating the output of the image intensifier to the input of the CCD at the same size. This type of coupling is not only the most efficient possible, but lens effects such as vignetting are eliminated. For some CCDs and image intensifier combinations not all of the CCD area will be illuminated by the image intensifier. The outlines of two different CCDs and two different image intensifiers are shown in Figure 3. As can be seen from the diagram, the combination of an EEV CCD and an 18 mm image intensifier results in the entire area of the CCD being illuminated. The combination of the same image intensifier and the EEV CCD array results in only about 17.5 mm being illuminated along the center of the CCD. Two detectors, the ICCD-576LD-E and the ICCD-512EFT, use 1.5:1 fiber optic tapers to transfer the light from the image intensifier to the CCD. Figure 3 is not applicable to these detectors. Figure 3. Size comparisons for 1:1 coupling 25 mm intensifier 18 mm intensifier EEV CCD EEV CCD The CCD is seated on a cold finger that is in turn seated on a thermoelectric (Peltiereffect) cooler. Both the image intensifier and the CCD are open to the atmosphere: no vacuum is present in these detectors. Because these components are cooled, nitrogen gas flow is used to prevent condensation. Figure 2 shows the nitrogen inlet for two different variations of the ICCD. Nitrogen flushing of the detector is described in Chapter 3. The thermoelectric cooler is mounted to the heat removal block. Heat is carried away from the Peltier device by water or other coolant. The coolant ports shown in Figure 2 are for connecting the coolant supply. See Chapter 3 for a complete description.

10 10 ICCD Detector manual Version 3.F The rear enclosure contains the preamplifier, the array driver, and the HV power supply electronics. This keeps all signal leads to the preamplifier as short as possible, and also provides complete RF shielding. In ICCD models with the FG-101 option, this enclosure also contains gating circuitry. See the separate FG-101 section in Chapter 6. The rear panel, with or without the FG-101 Option, is shown in Figure 4. Figure 4. View of standard and FG-101 ICCD detectors from the rear Standard ICCD Detector DB25 connector (to controller) ICCD with FG-101 Option Intensifier gain potentiometer BNC for high voltage from pulser: 50 Ω termination for FG-100, 1 MΩ for PG-200 or PG-10 Gate/shutter switch DB9 connector (to delay generator) BNC for 5 V TTL logic pulse The rear panel of a standard (non FG-101) ICCD will usually include a label indicating the allowable gate pulse voltages. A blue label (220 V) indicates a Gen II intensifier while a green label (900 V) indicates a Gen IV intensifier. An option is available from PI that allows the Shutter/Gate switch and the intensifier gain to be controlled remotely. The intensifier gain can also be controlled by an analog signal ranging from 0-10 V, corresponding to the 0-10 setting of the Intensifier Gain Potentiometer. Any of these options require factory modification. Contact your local sales representative for details. Preamplifier Selector switch ICCDs with the Thomson chip have two switch-selectable preamplifiers. The Preamplifier Selector switch is located on the back panel as shown in Figure 5, allowing performance to be optimized for two different A/D converters. Setting the switch to the H (up) position selects high-speed 1 MHz operation. Setting it to the S (down) position optimizes performance for the slower but quieter A/D, which can range in speed from 50 khz to 500 khz. The switch is recessed so that it can t be accidentally operated. Users are advised to use a small screwdriver to change the switch setting.

11 Chapter 1 General Information 11 Note that the switch setting also reverses the direction of data readout. If the switch is in the S (slow) position, data readout begins with the longer (red) wavelength pixels and progresses to the shorter (blue) wavelengths. If set to the H (fast) position, the shorter wavelength pixels are read out first. This reversal can be readily accommodated using WinSpec/32 s Hardware Setup Display Reverse feature, or by rotating the detector on the spectrometer. Figure 5. ICCD with Thomson chip and Preamplifier Selector switch ICCD Rear Panel H S H for fast (1 MHz) Preamplifier Selector Switch S for HI Resolution (50 khz to 500 khz)

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13 Detector Setup Chapter 2 This chapter covers general instructions and explains the different operation modes of the ICCD detector. Connection procedures for both imaging and spectroscopic ICCD systems are also explained. General Instructions Whether the ICCD is to be operated in a gated or CW mode, the proper connections must be made between the controller and the detector and the controller may need its internal switches set. Connecting the detector Each detector is supplied with a 25-pin cable to connect to the controller. Make sure that the controller is off, then connect the larger end of the cable to the port marked detector on the controller. Tighten the screws in place. Connect the smaller end of the cable to the rear panel of the detector and tighten the screws. Setting the controller Any user who has purchased both an ICCD and an LN/CCD (liquid nitrogen cooled) detector must ensure that the internal power supply switches of the controller are set properly for the ICCD. Consult the controller manual for instructions on setting these switches. If you have purchased only a single ICCD detector or an ICCD and a TE/CCD detector the controller is configured at the factory, and no user adjustments are needed. Electronic Control of the Image Intensifier The image intensifier of the ICCD serves as both a fast, electronic shutter for the incident light and as an amplifier for the light signal. Turning the image intensifier on and off can be accomplished in two ways, depending on the exposure time required. The switch to change between these modes is found on the back panel of the ICCD detector, as shown in Figure 4. In Shutter (sometimes called CW) mode the controller turns the image intensifier on and off. The exposure time is set in the application software, and no pulser is used. This mode is for exposures of a few tens of microseconds to DC (slower for some older detectors). In Gate mode, the image intensifier is turned on and off with high voltage from a PI Gate Pulse Generator. Four models of gate pulse generators are available, the fastest of which can generate pulses with a FWHM of about 2 nsec. 13

14 14 ICCD Detector manual Version 3.F The FG-101 has built in gating circuitry available for the ICCD detector. Its presence can be determined by comparing the back panel of your ICCD with those shown in Figure 4. For operation with the FG-101, the detector is set to Shutter or Gate mode, but no high voltage pulser is necessary. Shutter (CW) operation CAUTION When the detector is operated in Shutter mode the Gate BNC connector on the back of the detector should be covered to prevent accidental shock. CW operation requires only the ICCD and the controller to function. No pulser is necessary. The image intensifier can be turned on and off in as little as 50 µsec for ICCD detectors shipped after July, Detectors shipped before this date may only shutter down to 5 msec with the controller. When the ICCD is set to shutter mode, the image intensifier is Off until the experiment begins. Once the experiment begins, the image intensifier is turned On for the exposure time set in the software. After the exposure the image intensifier is turned Off, and the readout of the CCD array begins. This operation is notably different from the CW mode of PI intensified diode array systems. In the intensified diode array system, CW mode meant that the intensifier was turned on whenever the controller was powered. Because the Shutter mode results in longer exposure times, the likelihood of overexposure and damage to the intensifier is increased. If no pulser is connected and you are leaving the system powered on but unattended turn the switch to Gate. Operation with a PI Pulser and a Gen II Intensifier WARNING Detectors with the FG-101 option should never be connected to a high voltage pulser. High voltage pulses will permanently damage these detectors. RS Princeton Instruments (PI) cannot take responsibility for ICCD detector damage due to misuse. In Gate mode, standard ICCD detectors (those without the FG-101 option) require high voltage pulses from a PI Gate Pulse Generator to turn the image intensifier on and off. PI offers three high voltage pulsers, depending on the application and the timing it requires. The PI models are the PG-200, the FG-100, and the PG-10. CAUTION If you have a gate pulser from another manufacturer, contact RS Princeton Instruments (PI) before attempting to gate the ICCD with it. Failure to terminate the pulser correctly may cause damage to your equipment. All ICCDs purchased in combination with either the model PG-200 or the model PG-10 Pulser are internally terminated with a 1 M resistor before shipping. Detectors purchased with the model FG-100 Pulser are internally terminated in 50 at the factory. Customers who purchased only a single ICCD detector and a single pulser as part of the same system need not worry about the termination, as this is factory set. If you have more than one detector or pulser and are unsure of the impedance of the detector, you can measure it by connecting a ohmmeter across the Gate connector.

15 Chapter 2 Detector Setup 15 WARNING Operating the ICCD with the Gate BNC improperly terminated may cause permanent damage to the ICCD detector and the pulser. Damage due to misuse is not covered by warranty. To operate a 1 M-terminated detector (one originally purchased with a PG-200 or PG-10) with an FG-100 you have to attach a 50 (5 watt) terminator, available from PI, between the Gate (BNC) port of the detector and the cable from the pulser. Do not connect this terminator to the back of the pulser. To operate an ICCD internally terminated with 50 (one originally purchased with an FG-100) with a PG-200 or a PG-10, follow the instructions in Appendix C. ICCD detectors shipped with an FG-100H Pulser (a variation of the standard FG-100 Pulser) are shipped with an external 75 terminator. See Appendix C for details about this terminator. To connect the pulser to the ICCD, first make sure the detector is in Gate mode, and that the pulser is turned off. Connect the High Voltage Gate Out BNC on the back of the pulser to the Gate BNC on the back panel of the ICCD. Do not turn on the pulser. Initial operation will use signals generated by the controller. Other connections will need to be made from the controller to the pulser and from the pulser to the experiment. See the pulser manual for more details. The fastest gate possible with an ICCD depends on the speed of the pulser and the speed of the image intensifier. The PG-200, for example, can provide a 5 nsec gate pulse to the image intensifier, but the image intensifier may not be capable of gating at this speed. The gating speed of the image intensifier is generally specified at the time of order. If you are not getting the gating you expect first check the experiment timing, then contact the factory for more information. Operation with a Gen IV Intensifier Operation with a Gen IV Intensifier requires a higher voltage pulser, specifically a Berkeley Nucleonics Corp. Model 6040 Universal Pulse Generator equipped with a Model 310 plug-in Electrical Module. The pulser is modified at PI for operation with an ICCD and so must be obtained from PI. Additionally, a Tee connector, adapters and a high power external 50 terminator are required. These are provided when a system centered on an ICCD with a Gen IV Intensifier is purchased. As previously described, the ICCD Gate In connector is a type BNC connector in standard units. This is also the case for early ICCDs equipped with a Gen IV Intensifier. However, in newer ICCDs having a Gen IV Intensifier, Gate In is a type SHV connector. The SHV connector helps assure that the connections will be made properly and provides better safety in high-voltage applications. Although BNC and SHV connectors are superficially similar, they are not compatible. Cables used to connect to them must be fitted with the correct type connector for connections to be established. The connections are made as follows. Connect a Tee connector (BNC or SHV according to the Gate In connector type, to the to the Gate In connector of the ICCD. Be sure the Pulser power is OFF.

16 16 ICCD Detector manual Version 3.F Connect a cable from one side of the Tee to the HV output of a Berkeley Nucleonics Corp. Model 6040 Universal Pulse Generator equipped with a Model 310 plug-in Electrical Module. This cable should be no more than 1 m long. A type N-to- BNC or SHV adapter, whichever is required, will be needed to make the connection. Any required adapters would normally be supplied with the system. Connect a second cable from the other side of the Tee to the high-power 50 terminator. This cable should also be no longer than 1 m. CW or Shutter Operation WARNING To prevent equipment damage, the Berkeley Nucleonics Corp. Pulser must be disconnected for CW/Shutter mode operation. Operation with the FG-101 ICCD detectors can be purchased with internal gating circuitry, the FG-101 option. To check for the presence of this option, compare the rear panel of your detector with Figure 4. Detectors with the FG-101 option have a 9-pin connector not found on standard models. If you have this option, never connect high voltage from a gate pulse generator to the Gate port. The Gate port on these detectors is suited for TTL pulses only. See Chapter 6 for a description of how to correctly connect the FG-101 for pulsed operation. Imaging Applications This section describes how to connect lenses to the detector for imaging applications. Instructions for spectroscopic applications appear later in this chapter. Figure 6. Nikon lens adapter Screws for mounting lens adapter Set screws (4) to lock front part of adapter in place Lens release lever Front part of adapter for adjusting focus

17 Chapter 2 Detector Setup 17 Connecting lenses Detectors for use in imaging systems (cameras) are shipped with the lens mount already attached. Standard PI lens mounts use the Nikon bayonet format, as shown in Figure 6. This can be converted to most other formats using commercially available adapters. If your optical system cannot be converted to this format, contact the factory. Other mounts, including Canon and C-mount, are also available. Instructions for connecting these lenses are similar. To mount the lens on the camera, locate the large indicator dot on the side of the lens. There is a corresponding dot on the front side of the adapter. Line up the dots and slide the lens into the adapter. Turn the lens counterclockwise until a click is heard. The lens is now locked in place. If the front part of the lens mount rotates with the lens, tighten the set screws until it is fixed in place. Fine adjustments of the front part of the adapter are covered in Chapter 4. To remove the lens, locate the lens release lever at the front of the lens mount. Press the lever toward the camera housing, and at the same time rotate the lens clockwise. Then pull the lens straight out. Many standard microscope adapters are also available through PI. Attach the adapter to the lens mount provided with the detector. See Chapter 5 for additional information. Note: C-mount detectors are shipped with a dust cover lens installed. Although this lens is capable of providing surprisingly good images, its throughput is low and the image quality is not as good as can be obtained with a high-quality camera lens. Users should replace the dust-cover lens with their own high-quality laboratory lens before making measurements. Overexposure protection WARNING Image intensified detectors (ICCDs), can be destroyed if exposed to excessive light levels. RS Princeton Instruments (PI) cannot take responsibility for ICCD damage due to misuse. ICCDs must not be continuously exposed to high level radiation ( 10-4 foot candles). When the illumination level is not quantitatively known, set the lens to the smallest aperture (highest f-number) and place neutral density filters in front of the camera. If the experimental conditions dictate that only a small portion of the photocathode is illuminated over relatively long periods of time, change the illuminated region of the photocathode periodically to avoid long term localized photocathode or MCP damage. Cover the lens with a lens cap and continue with the instructions in Chapter 3. Spectroscopic Applications The ICCD detector must be properly mounted to the spectrometer to achieve the highest resolution. In the correct orientation the water ports will be located on the sides of the detector and the text on the back of the detector should be right side up.

18 18 ICCD Detector manual Version 3.F For spectrometers with a focal plane 25 mm or more beyond the exit interface, the detector adapter will be in two pieces, similar to those shown in Figure 7. These spectrometers include the ISA HR320, ISA HR640, Chromex 250IS, and most instruments that are 1 meter or longer. (If you are not sure of the depth of the exit focal plane, contact the spectrometer manufacturer.) Figure 7. Deep focal plane adapters Set screw Flange 1 Flange 2 Bolt flange 2 to the detector using the screws provided. Loosen the set screws on flange 1, then mount this flange to the spectrometer. Slide flange 2 into flange 1. Do not tighten the set screws until focusing and alignment are accomplished in Chapter 4. For spectrometers with a focal plane distance less than 25 mm, no adapter is used. Focusing is accomplished by adjusting the spectrometer. To attach the detector to the spectrometer, first screw the hex screws halfway into three of the six tapped holes in the spectrometer s exit plane. Position the detector so the three hex head screws line up with the openings in the detector. Slide the detector over the screws and rotate into the proper orientation. Leave the detector free to rotate until it is aligned in Chapter 4. To reduce the incident light, cover and close the entrance slit completely. Model IVUV This model ICCD has an MgF window and an S-20 photocathode for operation in the nm spectral range. Both the detector and the spectrometer must be evacuated to detect wavelengths shorter than 200 nm. Mount the detector to the spectrometer, then consult the spectrometer manual for pumping instructions. WARNING The vacuum spectrometer should be pumped to a vacuum level of 10-5 to 10-6 Torr. Do not turn on the detector until you are sure this vacuum has been achieved. Failure to do so will destroy the image intensifier since medium range vacuum provides ideal conditions for HV discharges inside the detector nose.

19 Chapter 2 Detector Setup 19 System Connections Make the system interconnections (Figure 8) as follows before proceeding. Do not turn on the power until directed to do so in Chapter 4. Figure 8. System connection diagram WARNING Always turn the power off at the controller before connecting or disconnecting the cable that interconnects the detector and controller or serious damage to the CCD may result. Damage due to misuse is not covered by the manufacturer s warranty. Camera to Controller cable: Each detector is supplied with a 25-pin ICCD cable to connect to the controller. The cable and connectors are designed such that there is no possibility of connecting them incorrectly. In the case of an ST-130 or ST- 138 controller, connect from the DB-37 connector labeled Detector to the DB- 25 connector on the back of the camera. See the controller manual for additional information. Controller to Computer cable: This connection is made via the high-speed serial port. If the Controller is a Model ST-130 or ST-138, the serial port is located on the back of the controller. The system could additionally include a monitor or other system components that would have to be connected. See the controller manual for more detailed information.

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21 Flushing and Cooling Chapter 3 WARNING Never run the ICCD without water or coolant circulation. Running the ICCD cooled without circulating liquid can destroy the detector! The CCD array in an ICCD camera is cooled by a Peltier-effect thermoelectric cooler, driven by closed-loop proportional-control circuitry. A thermal sensor attached to the cooling block of the detector monitors its temperature. The coolant block is made of Delrin to prevent condensation, but this also prevents heat transfer from the thermoelectric cooler to the atmosphere. Water or other coolant is therefore necessary to carry away heat generated by the thermoelectric cooler. ICCDs typically have the following temperature ranges: -35 C with water circulation -45 C with chilled water circulation WARNING Do not cool below -45 C. This may damage the CCD or the bond between the fiber-optic window to the CCD. Nitrogen Flushing of the Detector All of the ICCD models described in this manual require continuous flushing of the detector with nitrogen or dry air while they are operating cooled. Even if the detector is not powered it must be flushed whenever water or other coolant is circulating, as the possibility of condensation still exists. WARNING CAUTION Never use any gas other than nitrogen or dry air. Noble gases, e.g., argon or helium, will destroy the image intensifier. If the relative humidity in your lab is high, e.g., > 40% do not operate the detector without flushing, even if you are not cooling the detector! The dryness of the nitrogen or air is critical. The colder the detector is operated, the dryer the gas must be. Also, the longer the detector is operated cold, the drier the gas must be. WARNING Be wary of unspecified dry gases intended for less demanding applications. Even a 99.9% pure air can deposit enough water to cause permanent damage to the detector. 21

22 22 ICCD Detector manual Version 3.F Consider as an example an ICCD camera where the CCD temperature is -35 C and where you are using air with 100 ppm of water vapor. This water vapor will be at dew point and water will condense and freeze on the CCD. Ice will continue to form for as long as this air flows over the surface of the CCD. PI recommends gases with less than 10 ppm water vapor be used. Some examples are AIRCO compressed air grades DRY (10 ppm) Zero 1.0 (3 ppm), and AIRCO compressed nitrogen grade 4.8 (3 ppm). Equivalent gases from other quality vendors are also suitable. Connect the nitrogen source to the nitrogen inlet. Flush the detector for at least 10 minutes at a high rate (1-2 liters/minute). Then lower the rate to ml/minute. For applications in high humidity climates, the exit port of the spectrometer may need to be continuously flushed with nitrogen. (It is a good general practice to regularly flush the spectrometer for condensation-free operation under all conditions.) CAUTION After you have finished an experiment and have turned off the controller, maintain a gas flow of at least 2 liters/minute for at least 30 minutes. This keeps condensation from forming on the detector until it reaches room temperature. PI recommends that the ICCD detector be returned to room temperature at least one hour out of every 24 for flushing. Coolant Connections WARNING Running the ICCD cooled without circulating liquid can destroy the detector! Two copper-and-brass fittings come with the ICCD. These connect to the two threaded coolant ports on the sides of the detector, and are a standard ¼ size. Connect these to the coolant supply, tightening firmly by hand. Either port may be used as the inlet. A closed circulator such as the PI model CC-100 or the Neslab model RTE-110 should be used for circulation of coolant other than water. WARNINGS Contact the factory before using any coolant fittings other than those supplied by PI. Forcing the wrong fittings onto the Delrin threads will destroy them. Take care that the coolant is ph neutral. Acidic or alkaline coolant can damage the detector fittings and internal cooling block through corrosion. Also, using hard water can cause mineral deposits to build up inside the fittings and cooling block. Damage from the use of unsuitable coolant could be very expensive to repair and is not covered by the Warranty. ICCD users may have also purchased the CPC-100 Photocathode Cooler option. If you have purchased this option, read the CPC-100 instructions at the end of this chapter. They contain precautions and handling instructions for this type of ICCD detector. Start the flow of coolant. A good initial value is ~1-3 liters/min.

23 Chapter 3 Flushing and Cooling 23 WARNING If you observe any coolant leaks, turn the controller off immediately. If the leak is due to external hoses or connections, replace those parts before continuing. Coolant leaking into the detector head or the electronics enclosure can permanently damage the ICCD. Setting the Temperature CAUTION Set the temperature of the detector, but do not turn the controller on at this time. Setup precautions are covered in detail in Chapter 4. Locate the Temp knob on the front of the controller. The dial reads in units of minus degrees Centigrade. See Figure 9 to locate the Locking tab. Push this tab counterclockwise until the Temp knob is free to rotate. WARNING The lowest operating temperature permitted with an image intensified CCD detector is -45 C. Lower temperatures may damage the image intensifier tube or the optical coupling between it and the CCD. Figure 9. Temp. knob, located on the front of the controller 100 C 1 C (unlocked) (locked) locking tab On the top side of the Temp knob is a rectangular window that denotes hundreds of C. Each complete turn of the knob is -100 C. Around the moveable part of the knob are numbers from 0 to 99, in increments of 2. Turn the knob until the correct value (0) appears in the hundreds box. Then turn the knob until the desired value between 0 and 45 appears below the box. Turn the locking tab clockwise to lock the Temp knob in place. Figure 9 shows the position of the dial for a temperature of -40 C.

24 24 ICCD Detector manual Version 3.F CPC-100 Photocathode Cooler Option WARNING Care must be taken when handling the CPC-100, as the coolant connections on the nose of the detector are quite fragile and are connected directly to the image intensifier. Banging these connectors or using excessive force can permanently damage the image intensifier. Figure 10. Do not bang the connections to the CPC-100! Connections and operation The CPC-100 Photocathode Cooler option allows chilled liquid to cool the photocathode of the image intensifier. This cooling significantly reduces the EBI noise of the image intensifier, just as cooling the CCD array reduces its noise. An ICCD with the CPC-100 option is shown in the bottom half of Figure 2. The CPC-100 Photocathode Cooler requires circulation of a low temperature coolant, such as 50% mixture of ethylene glycol and water, at a temperature of -20 to -30 C. Because the nose of the ICCD detector has no feedback system to ensure a constant temperature of the image intensifier photocathode, it is important to use a coolant refrigerator that can provide very precise thermostating. One example is the Neslab model RTE-110, which has ±0.010 C thermostating precision at temperatures as low as - 25 C.

25 Chapter 3 Flushing and Cooling 25 WARNINGS If you are using a detector with a CPC-100 cooler on a spectrometer you must flush the spectrometer with nitrogen to prevent condensation on the intensifier s window. This is because the entire intensifier is cooled to approximately 12 C. Water from the air condensing on the intensifier can destroy it. RS Princeton Instruments (PI) cannot take responsibility for detector damage due to misuse. Hoses are shipped already connected to the CPC-100 cooler. Do not snap or force these coolant hoses. If you observe any coolant leak, turn the controller off immediately and contact the factory. Figure 11. Making connections to the CPC-100 Push Gently water hose foam insulator These CPC-100 coolant hoses connections are normally done at the factory before the unit is shipped. If the hoses need to be reattached, gently connect the supplied coolant hoses to the barbed CPC-100 connections, as shown in Figure 11. Do not use excessive force to attach these hoses, as the barbed connectors are rigidly fixed to the image intensifier. Slide the provided foam insulator around the coolant hoses as shown. WARNING Any jarring of these connectors, even resting the detector on these barbed connectors, can damage the image intensifier. Note: If you ever need to disconnect the hoses from the CPC-100 it is probably safest to cut the hoses off below the ends of the barbs. The remaining pieces of tubing can be cut open lengthwise and removed. The barbed shape of the connectors, although effective in preventing leakage, is not easily disconnected.

26 26 ICCD Detector manual Version 3.F Connect the refrigerator to the copper tubes (¼) at the other end of the coolant hoses. The preferred mode of operation is to connect the refrigerator only to the CPC-100. Separate water hoses are usually attached to the coolant connections on the body of the detector, as shown in Figure 12. The temperature of the CCD is electronically regulated to within about ±0.050 C, so chilled coolant at a specific temperature is not required. If you wish to use the refrigerator to cool the CCD also, connect the two in parallel, with the majority of the coolant going to the photocathode. Figure 12. Standard CPC-100 coolant connections to water flow Nitrogen Gas Inlet Coolant Port Tubing and insulator Clamp Copper tubing Retrofit If you would like to add the CPC-100 option to an existing full frame ICCD-576 detector, contact the factory for pricing and delivery schedules. The detector must be returned to the factory for this modification.

27 Focusing Chapter 4 Detectors for either imaging or spectroscopic applications must be focused for maximum resolution. Imaging applications require adjustment of both the lens and the lens adapter. Spectroscopic applications require focusing and alignment of the spectrometer. Before proceeding with the focusing procedures, there are some equipment and personnel safety issues that need to be addressed. System Connections Verify that the system interconnections have all been made before proceeding. Camera to Controller cable Controller to Computer CAUTION Do not connect the ICCD to the pulser at this time. If the ICCD is in Shutter mode and the pulser is connected the HV circuitry in the detector head will in effect be driving the pulser, and diminished performance will result. The system could additionally include a monitor or other system components that would have to be connected. See the controller manual for more detailed information. Precautionary Measures WARNING Image intensified CCD detectors (ICCDs) can be destroyed if continuously exposed to light levels higher than twice the A/D saturation level. To prevent damage to the detector, check that the following conditions are met before turning on the controller. Laboratory conditions Check that all the following conditions are met before operating your ICCD camera. The temp knob is set to a temperature of -45 C or higher The detector is set to the Gate mode Any HV pulsers present are turned off The detector is completely blocked off Nitrogen gas is flowing through the detector Water or other coolant is circulating 27

28 28 ICCD Detector manual Version 3.F WARNING Condensation To prevent damage to the camera, take immediate corrective action if condensation develops anywhere on the exterior of the camera. Corrective action includes immediately raising the set temperature and/or lowering the humidity in the room. Image intensifier gain The Intensifier gain potentiometer on the rear panel of the detector provides a continuous variation in the detector gain. This gain is adjustable from approximately 1-70 counts/photoelectron on ICCDs with 1:1 fiber coupling. Note: Versions of the ICCD shipped after June, 1996 use a new version of the high voltage board within the detector. The Gain knob on these detectors is effective throughout the entire range of the knob, from Older ICCD detectors were only effective from a setting of about 8-10 on the dial. Locate the Gain knob on the rear panel of the detector (see Figure 13). Turn the locking tab counterclockwise until the knob is free to rotate. Set the gain of the ICCD detector to between 9 and 10. The box at the top of the Gain knob should read 9, as shown in Figure 13. CAUTION If the gain of the intensifier is set at a low level, i.e., below approximately 20 counts/photoelectron (about 2 on the dial), protective circuitry within the detector becomes less effective. Figure 13. Intensifier gain adjustment (unlocked) locking tab (locked) Adjusting the intensifier gain also affects the dark charge of the intensifier (EBI). To properly perform background subtraction a background must therefore be measured at each intensifier gain setting. Adjustment precautions Once the controller is turned on (described in the section below) the gain can be adjusted any time during image acquisition. For systems shipped before July, 1996, it is best to adjust the knob from lower to higher values. If you must adjust the gain to lower values on these detectors, do so slowly. Please also see the caution below. CAUTION If the Image Intensifier Gain knob of an ICCD shipped before July, 1996, is turned from higher to lower values too quickly the photocathode may turn on momentarily. This is normal operation for this detector, and will not harm the detector as long as the incident light levels do not exceed acceptable values.

29 Chapter 4 Focusing 29 Alarm To reduce the risk of detector damage, ICCD detectors shipped after July, 1996, are equipped with an audible alarm in the detector head, activated when the intensity of light falling on the image intensifier exceeds a preset threshold. While the alarm is sounding the photocathode and MCP power are temporarily disabled. The detector window must be immediately covered or the controller must be turned off until the illumination level is readjusted. If the alarm sounds continuously even when the illumination level is adequately low, its threshold may have to be internally readjusted at the factory. The ST-138 and ST-130 controllers also provide an audible alarm, indicating the same over-threshold condition as the detector alarm. Note that it is normal for the alarm to sound momentarily when the system is turned on. CAUTION Discontinue operation and contact the factory at once if sporadic or continuous unwarranted alarms occur. They may indicate intensifier damage or another situation that requires immediate attention. Power Up Turn on the power to the controller and the computer. If the controller is an ST-130 or ST-138, check the Cooler switch. If it is Off, set it to On. After the cooler switch is turned on, it takes from minutes for the ICCD to reach its preset temperature. The cooler status indicator (ST-130 or ST-138 controller) will turn from orange to green to indicate that the temperature is thermostated to within ±0.050 C. Note: Temperature regulation does not reach its ultimate stability for at least 30 minutes after temperature lock is indicated. Also note that, since the thermoelectric cooler has no ability to heat the CCD, the detector will be more thermally stable at lower temperatures. Continue with the focusing instructions applicable to your application. Baseline Signal With the detector completely blocked, the CCD will collect a dark charge pattern, dependent on the exposure time, detector temperature, and intensifier gain setting. The longer the exposure time and the warmer the detector the larger and less uniform this background will appear. Leave the detector in the Gate mode for dark charge readings. After the green status light for the cooler has lit, wait 30 minutes for the detector temperature to completely stabilize. Then try taking a few dark charge readings.

30 30 ICCD Detector manual Version 3.F Note: Do not be concerned about either the baseline level of this background or its shape, unless it is very high, i.e., > 1000 counts. What you see is not noise. It is a fully subtractable readout pattern. Each CCD has its own dark charge pattern, unique to that particular device. Every device has been thoroughly tested to ensure its compliance with PI s demanding specifications. CAUTION If you observe a sudden change in the baseline signal you may have excessive humidity in the detector enclosure. Turn off the controller and increase the flow of nitrogen through the detector. After minutes, resume operation. When you have finished taking dark charge data, disconnect the pulser set the detector to the Shutter mode. Shutter vs. Gated Operation In either Gate or Shutter mode a positive bias voltage is applied to the intensifier photocathode in the off state. When the intensifier is turned off the detector is practically blind, with an on/off ratio of approximately :1 (the exact ratio depends on the gain setting). The Gate mode of standard ICCD detectors (detectors without the FG-101 option) requires a HV signal from a pulser to trigger the image intensifier. The detector is turned on while the gate pulse is being received through the BNC connector on the back of the detector. Detectors with the FG-101 option require only a TTL logic pulse for gating. See Chapter 6 for complete details of this option. Note: The on-off ratio of the intensifier is excellent ( typical). Even so, there may be situations where this may not be sufficient to prevent significant signal leakage when the intensifier is gated off. That this is so becomes apparent when the gate on/off time factors are considered. In an experiment with a narrow gate pulse and slow readout rate, the ratio of the off time to the on time may easily be of the same order as the on/off ratio of the intensifier itself. Where this is the case, the signal impinging on the CCD during the intensifier off time may easily equal the signal applied during the intensifier on time. The effect of this signal will be to cause data smearing. The only solution to this problem at this time, other than to increase the pulse rate or the gate time to where the effect is insignificant, is to mount an external shutter ahead of the intensifier. Future intensified detectors may incorporate design advances to assure that this problem never occurs. In the Shutter mode a signal from the controller causes the image intensifier to gate for a certain time interval. Then the intensifier is turned off once again. Note that in operation with the ICCD, the controller waits about 6 ms after completion of the exposure before starting the readout. CAUTION Do not connect the ICCD to the pulser if operating in Shutter mode. If the ICCD is in Shutter mode and the pulser is connected the HV circuitry in the detector head will in effect be driving the pulser, and diminished performance will result. Since the Shutter setting is controlled internally, use it for the tests below. Once data collection is confirmed turn off the controller, connect the HV pulser to the detector, and switch to the Gate mode. Once again, detectors with the FG-101 option should never be connected to a HV pulser.

31 Chapter 4 Focusing 31 Focusing Imaging Systems Begin with the lens blocked off. Set the lens at the smallest possible aperture (largest f-number). As a guide, light barely visible to the human eye is a good starting level for shutter mode operation. Set the software to the Freerun and Asynchronous modes (consult the software manual for details). Choose a short exposure and begin data collection. If gating, see the pulser manual to establish proper gating. It is usually easier to get the system running in Shutter mode first, then switch to Gated mode. Slowly uncover the lens. If the alarm sounds, cover the lens quickly, and reduce the light level. Adjust the exposure until a suitable value is found. Check the brightest regions of the image to determine when the full scale of the A/D converter is being used. Place a suitable target in front of the lens. Objects with text or graphics work best. The target should be located at infinity (at least 200 focal lengths from the lens) if possible. If not possible set the focus ring of the lens to the actual distance to the target and continue. Reduce the light and/or exposure to allow the operating lens iris to maximum or near maximum aperture without overloading the camera. Using a large aperture, set the focus adjustment of the lens to infinity. Loosen the lens mount set screws with a Allen wrench, While observing the image at the computer, rotate the lens and the front part of the adapter as a unit until the sharpest possible focus is obtained. Tighten the set screws. All focusing may now be done with the focus adjustment on the lens. Imaging field of view When used for two-dimensional imaging applications, PI CCD cameras closely imitate a standard 35 mm camera. Since the CCD is not the same size as the film plane of a 35 mm camera, the field of view at a given distance is somewhat different. Figure 14. Imaging field of view Object Lens Intensifier O S D B D = distance between the object and the image intensifier B = 46.5 mm (Nikon bayonet only)

32 32 ICCD Detector manual Version 3.F F = focal length of lens S = horizontal or vertical dimension of CCD O = horizontal or vertical field of view covered at a distance D M = magnification The field of view is: O = S M, where M = FD ( D B) 2 Spectroscopy Systems Note: If you purchased an optical-fiber adapter and cable, install them only after the regular alignment procedure has been successfully completed. Consult the Optical Fiber Adapter manual for specific instructions. Set the spectrometer entrance slit width to minimum (10 µm if possible). Place a standard emission light source (ideally a Hg pen-ray lamp) in the range of m away from the entrance slit. Set the software to the Freerun and Asynchronous modes (consult the software manual if you are unfamiliar with these modes). If gating, see the pulser manual to establish proper gating. It is usually easier to get the system running in Shutter mode first, then switch to Gated mode after the optics are adjusted. Begin data collection. Rotate the spectrometer grating to approximately the zero order counter position. As you rotate the grating, a line should be seen moving across the screen. Remember that the image on the screen is not the last frame taken, since each frame may take 2 seconds or more to be processed. Do not move the spectrometer grating so fast that the zero order line is missed. Rotate the grating to the first (or second) order counter position and check the illumination of the entrance slit. If a lens is used to focus the lamp s light, try to match the f-number of the spectrometer in order to fill up the grating as uniformly as possible. Observe an image of the spectrum. Spectral lines should be absolutely vertical or horizontal. If the lines do not run horizontally or vertically, slowly rotate the detector until spectral lines become aligned. Focusing the detector is achieved differently on different spectrometers. On models where the adapter is made of two pieces that slide together, focusing is achieved by slowly sliding the detector in and out of the exit focal plane. One-piece adapters rely on a focusing adjustment on the spectrometer. See the spectrometer manual for details.

33 Chapter 4 Focusing 33 Check the intensity of pixels on either side of a peak. For well resolved spectral lines, this intensity should be approximately 75% of the peak intensity. If the spectral lines are not this well resolved, first check the focus. Then try rotating the detector with respect to the spectrometer. WARNING When optically aligning the detector make sure you do not accidentally expose it to room light. Alignment and focusing may need to be repeated for optimal alignment. When completed, tighten the screws that attach the adapter to the spectrometer to prevent accidental misalignment. To tighten the set screws on the two-piece adapter, first stop data collection. The top of the spectrometer may need to be removed to access the set screws. Kinetics ICCD with Moveable Mask In kinetics operation, an image is projected on a narrow strip at one end of the image intensifier and the rest of the intensifier is masked and used as a storage area. If the open area is small relative to the storage area, quite a few images can be acquired in rapid succession, for example with 50 open rows and 256 rows masked, 10 images can be acquired in a total of approximately 1.2 msec. To do this on an ICCD, the photocathode on the intensifier must be masked. To create an intensified camera with the most flexible kinetics mode possible, a moveable mask was required. To minimize the effective distance between the intensifier photocathode and this mask, a fiber optic window is used on the input of the intensifier. Precision screws adjust the mask from no coverage (full frame) to half coverage or nearly full coverage (kinetics). PI ICCD detectors with a moveable mask are capable of kinetics operation when used with a Model ST-138 Controller. For detectors shipped after June, 1996, gating is accomplished using any standard pulser or the FG-101 option. Kinetics detectors shipped before this time required the FG-101 option. Because of the mechanical requirements of the moveable mask, the voltages on this type of ICCD are different from the standard ICCD. This makes the performance specifications somewhat different. See the Specifications beginning on page 44 for details. This detector may also use a Thomson 576 x 384 CCD. If you have purchased a detector with this capability, follow the directions below to adjust the position of the mask. Note: A small amount of light leakage may occur through the holes for the adjustment screws, even when plugged. After adjustment is complete a piece of electrical tape or similar opaque material can ensure total light shielding. The two adjustment screws shown in Figure 15 are normally plugged to prevent light leakage. Remove the plugs for the duration of the adjustment.

34 34 ICCD Detector manual Version 3.F The adjustment screws are 8-80 hex head screws, and can be adjusted with a 5/64 Allen wrench (one is provided with this system). For initial collection, open the mask so at least a third of the image intensifier is visible. Collect a full frame image with the system (kinetics and frame transfer disabled in the software). Determine how many pixels the mask must be moved. When adjusting the mask, do not apply any significant torque to the screws, as the mechanical mask can damage the intensifier if pushed beyond its allowable limit. Figure 15. ICCD with kinetics option, viewed from below Adjustments for moveable mask To adjust the mask, turn each screw no more than one turn before turning the other screw by the same amount. This keeps the mask from tilting at an angle as it moves. For and ICCDs one full turn corresponds to approximately 14 pixels. For ICCDs one full turn corresponds to approximately 12 pixels. To reduce the image area, turn the screws clockwise. To enlarge the image area, turn the screws counterclockwise. For most frame transfer applications the mask should fall in the exact center of the array. To level the mask with respect to the CCD turn the screws small, equal amounts in opposite directions. You can easily determine the deviation from level by zooming in on an image of the mask edge. With careful adjustment, tilt can be reduced to ±1 pixel.

35 Microscopy Applications Chapter 5 Introduction This chapter discusses the setup and optimization of your digital imaging system as applied to microscopy. Since scientific grade cooled imaging systems are usually employed for low light level microscopy, the major goal is to maximize the light throughput to the camera. In order to do this, the highest Numerical Aperture (NA) objectives of the desired magnification should be used. In addition, you should carefully consider the transmission efficiency of the objective for the excitation and emission wavelengths of any fluorescent probes employed. Another way to help maximize the transmission of light is to choose the camera port that uses the fewest optical surfaces in the pathway, since each surface results in a small loss in light throughput. Often the trinocular mount on the upright microscope or the bottom port on the inverted microscope provides the highest light throughput. Check with the manufacturer of your microscope to determine the optimal path for your experiment type. A rule of thumb employed in live cell fluorescence microscopy is if you can see the fluorescence by eye, then the illumination intensity is too high. While this may not be universally applicable, it is a reasonable goal to aim for. In doing this, the properties of the CCD in your camera should also be considered in the design of your experiments. For instance, if you have flexibility in choosing fluorescent probes, then you should take advantage of the higher Quantum Efficiency (QE) of the CCD at longer wavelengths. Front illuminated CCDs generally reach peak QE between 600 and 800 nm. Hardware binning can also be used to increase sensitivity. If sufficient detail will be preserved, you can use 2 2 binning (or higher) to increase the light collected at each super-pixel by a factor of four or more. This will allow the user to reduce exposure times, increasing temporal resolution and reducing photodamage to the living specimen. Another method to minimize photodamage to biological preparations is to synchronize a shutter on the excitation pathway to the internal shutter on the camera. This will limit exposure of the sample to the potentially damaging effects of the excitation light. Mounting the Camera on the Microscope The camera is connected to the microscope via a standard type mount coupled to a microscope-specific adapter piece. There are two basic camera mounting designs, the F- mount and the C-mount. The F-mount uses a tongue and groove type mechanism to align the camera with an adapter, while the C-mount employs a standard size thread to connect to the adapter. Either or both types could be available for a specific camera model. 35

36 36 ICCD Detector manual Version 3.F F-Mount For a camera with the F-mount type design, you will need two elements to mount the camera on your microscope. The first element is a Diagnostic Instruments Relay Lens. This lens is usually a 1 relay lens, which performs no magnification. Alternatively, you may use a 0.6 relay lens to partially demagnify the image and to increase the field of view. There is also a 2 relay lens available for additional magnification. The second element is a microscope specific Diagnostic Instruments Bottom Clamp. Table 1 shows which bottom clamps are routinely used with each of the microscope types. They are illustrated in Figure 16. If you feel that you have received the wrong type of clamp, or if you need a clamp for a microscope other than those listed, please contact PI. Table 1. Bottom clamps for different microscope types Microscope Type Leica DMR Leitz All types Nikon Optiphot, Diaphot Olympus BH-2, B-MAX, IMT-2 Zeiss Axioscope, Axioplan, Axiophot Zeiss Axiovert L-clamp NLW-clamp O-clamp V-clamp Z-clamp ZN-clamp Diagnostic Instruments Bottom Clamp Type To assemble the pieces, first pick up the camera and look for the black dot on the front surface. Match this dot with the red dot on the side of the relay lens. Then engage the two surfaces and rotate them until the F-mount is secured as evidenced by a soft clicking sound. Now, place the long tube of the relay lens into the bottom clamp for your microscope, securing the two together with the three set screws at the top of the clamp as shown in Figure 17. This whole assembly can now be placed on the microscope, using the appropriate set screws on the microscope to secure the bottom clamp to the output port of the microscope. The F-mount is appropriate for any trinocular output port or any side port. When mounting the camera perpendicular to the microscope on the side port, we recommend that you provide some additional support for your camera to reduce the possibility of vibrations or excessive stress on the F-mount nose. PI does not advise using an F-mount to secure the camera to a bottom port of an inverted microscope due to possible failure of the locking mechanism of the F-mount. Contact the factory for information about a special adapter for operating in this configuration.

37 Chapter 5 Microscopy Applications 37 Figure 16. Diagnostic Instruments F-mount adapters 1X HRP 100-NIK Relay Lens L ZN O NLW Z V Bottom Clamps Figure 17. Bottom clamp secured to relay lens 1X HRP 100-NIK "L" bottom clamp

38 38 ICCD Detector manual Version 3.F Operation C-Mount For a camera equipped with a C-mount thread, use a standard C-mount adapter supplied by the microscope manufacturer to attach the camera to the microscope. If you don t have an adapter, you can obtain one through your microscope distributor or directly from Diagnostic Instruments in the US at Tel: The adapter can be screwed into the camera and then the assembly can be secured to the microscope using the standard set screws on the microscope. The camera can be mounted on the trinocular output port, the side port or the bottom port of the inverted microscope. When mounting the camera perpendicular to the microscope on the side port, we recommend that you provide some additional support for your camera to reduce the possibility of vibrations or excessive stress on the C-mount nose. For the bottom port of the inverted microscope, the C-mount is designed to support the full weight of the camera, however, you may wish to provide some additional support for the camera since the detector is in a position where it could be deflected by the operator s knee or foot. This kind of lateral force could damage the alignment of the nose and result in suboptimal imaging conditions. Most output ports of the microscope do not require additional optical elements to collect an image, however, please check with your microscope manual to determine if the chosen output port requires any relay lens. In addition, all optical surfaces should be free from dust and finger prints, since these will appear as blurry regions or spots and hence degrade the image quality. Xenon or Mercury Arc Lamp Precautions WARNING Before You Start, if your system includes a microscope Xenon or Hg arc lamp, it is CRITICAL to turn off all electronics adjacent to the arc lamp, especially your digital camera system and your computer hardware (monitors also), before turning on the lamp power. Powering up your microscope Xenon or Hg lamp causes a large EMF spike to be generated that can cause damage to electronics that are running in the vicinity of the lamp. We advise that you place a clear warning sign on the power button for your arc lamp reminding all workers to follow this procedure. While PI has taken great care to isolate its sensitive circuitry from EMF sources, we cannot guarantee that this protection will be sufficient for all EMF bursts.

39 Chapter 5 Microscopy Applications 39 Focusing the Microscope Imaging Hints Direct all of the light to the eyepieces Focus on the target, set up Koehler illumination and adjust the condenser to match the objective, all in the transmitted light mode (as per the instructions provided by your microscope manufacturer). Decrease the transmitted light intensity to a level that is low, but still sufficient to allow visualization by eye. Redirect the light to the camera port. Adjusting the Parfocality of the Camera To adjust the parfocality on an F-mount system, begin collecting images with a short exposure time and focus the light on the camera by rotating the ring on the Diagnostic Instruments relay lens without touching the main focusing knobs on the microscope. On a C-mount system, the camera should be very close to parfocal, although some C- mounts will be adjustable using set screws on the microscope to secure the adapter slightly higher or lower in position. While adjusting parfocality, you will need to acquire images rapidly to minimize the delay between the time a setting is changed and the time when the effect of the change can be observed. The specifics of how to proceed will vary according to the application software. In WinView, select Acquisition, Focus. Begin with an exposure time 0.1 sec and a Hardware LUT selection of Bit Range Then use RUN to begin data acquisition and STOP to end it when you are finished focusing. See your WinView manual for additional information. In IPLab, begin by selecting Set Camera from the Ext. menu. Set the dialog to Asynchronous, 100 msec exposure, and 2 cleans. Establish Focus mode operation by selecting Focusing from the Ext. menu. Set this dialog to 0.1 pixel seconds, gray scale, and 1. Press Start Camera to begin focusing and Stop Camera when finished. See your IPLab manual for additional information. Many PI cameras, both F-mount and C-mount, also make provision for extending the focus range by providing a focus adjustment on the camera lens mount. If necessary, this focus can be changed to bring the image into range of the relay lens or other microscope focus adjustment. Determine the gray levels of the image by placing the cursor within the image and monitoring the values shown. For optimal image quality of a 12-bit image, the highest value in the field should be near 4000 counts but not at 4095 (which is saturating). You may increase the number of counts by increasing your exposure or by increasing the amount of light illuminating the specimen.

40 40 ICCD Detector manual Version 3.F Fluorescence Once you have acquired a suitable image in transmitted light mode, you may switch to fluorescence mode. In fluorescence mode you generally want to minimize the bleaching of your sample, usually achieved by placing several neutral density filters in the excitation pathway to minimize the illumination intensity. There will always be a trade-off here; when you maximize signal quality by increasing the illumination intensity, you need to consider whether your preparation can tolerate these conditions. In general, it is better to expose longer with a lower intensity than to expose for a shorter time with a higher intensity; nevertheless, your experimental conditions will dictate which path you take. In fluorescence measurements you may not wish to maximize the gray levels in the image, since this may cause bleaching of the dye or photodamage to the cell. Maintain the minimum exposure required to get a sufficiently high quality image. If the scaling on the image does not appear good to the eye, you may use additional scaling features available in the software. See your software manual for information on how to properly use the contrast enhancing features of the program. Microscopes and Infrared Light Microscope optics have very high transmission efficiencies in the infrared region of the spectrum. Since the light sources are very good emitters in the infrared, some microscopes are equipped with IR blockers or heat filters to prevent heating of optical elements or the sample. For those microscopes which do not have the better IR blockers, the throughput of infrared light to the camera can be fairly high. In addition, while the eye is unable to see the light, some PI cameras are very efficient in detecting infrared wavelengths. As a result, the contaminating infrared light will cause a degradation of the image quality due to a high background signal that will be invisible to the eye. Therefore, it is recommended that you add an IR blocker prior to the camera if you encounter this problem with the microscope.

41 FG-101 Pulser Chapter 6 The FG-101 Pulser is a combination gate pulse generator and high voltage power supply built into the detector housing of an ICCD. Figure 4 compares the rear panel of a standard ICCD with the rear panel of an ICCD containing the FG-101 option. The ICCD with the FG-101 option has a 9-pin connector that is not present on a standard ICCD. WARNING Detectors with the FG-101 option should never be connected to a high voltage pulser. High voltage pulses will permanently damage these detectors. RS Princeton Instruments (PI) cannot take responsibility for ICCD detector damage due to misuse. The FG-101 provides gate pulses with widths from 5-6 nsec to DC, with pulse rise times in the 5 nsec range. Gating from 50 nsec to DC can be accomplished with a single TTL signal to the Gate port on the back of the detector. For gating shorter than 50 nsec two low-level pulses must be provided by a delay generator through the 9-pin connector on the rear of the detector, Both modes of operation are described below. Single TTL Input When the detector is in TTL mode and nothing is connected to the 9-pin connector, pulses are controlled with the Gate input on the back panel of the detector. The gating follows the TTL pulse, so the intensifier is on when the Gate signal is +5 V and off when the Gate signal is 0 V. Pulse widths can range from 50 nsec to DC, with some timing distortion present at shorter pulse widths. Delay Generator Input For pulses shorter than 50 nsec, the detector must be connected to a delay generator such as the Stanford Research Systems DG-135 or DG-535 Delay Generators. A special cable is available from PI to connect the ICCD to these delay generators. Contact the factory for more information. The 9-pin connector is used to control several detector functions using TTL. The CW pin can be used to override the Shutter/TTL switch on the rear panel of the detector, causing the detector to function in Gate mode. For pulse widths less than 50 nsec, the STSTEN pin must be enabled. Gating is then controlled by separate pulses to the START and STOP pins, with a propagation delay of approximately 5 nsec. For pulse widths of 50 nsec to DC either START/STOP or GATE can be used. Operation with the GATE pin is identical to Single TTL Input described in the section above. For operation with the DG-535 Delay Generator, the INHIBIT connector on the DG-535 should be connected to the SHUTTER MONITOR output of the controller. This will 41

42 42 ICCD Detector manual Version 3.F prevent the intensifier from gating while the CCD is being read out. Without this connection, smearing and high voltage spikes will occur in the data whenever the intensifier is gated at the same time that charge is being shifted out of the CCD array. At small pulse widths the timing distortion is proportionally greater, so the timing generator must be adjusted to compensate. This effect is most significant for pulse widths shorter than 30 nsec, and is negligible for pulse widths greater than 100 nsec. 9-Pin Connector Signals Figure pin connector for delay generator CW START GND STOP STSTACK GATE STARTRTN STSTEN STOPRTN 1 CW CMOS compatible, variable impedance input/output. High = Shutter (CW) mode, Low = Gate mode. 51 k if Shutter/TTL switch is in Shutter mode, 1 k if in TTL position. A remote device can sense state with high impedance CMOS input. Remote device can drive this pin with low impedance output, overriding the Shutter/TTL switch on the back of the detector. CW overrides STSTEN and GATE. 2 START input, negative edge sensitive (transformer coupled), 0 to +5 V, active if STSTEN True. Causes intensifier to switch to on state. Low impedance value is variable and complex, drivable from a 50 source. 3 GND Ground 4 STOP input, active if STSTEN is True. Causes intensifier to turn off. Drive characteristics similar to START. 5 STSTACK output, high impedance (51 k, 10 F) to +5 V, can be used by external device to sense if head is powered, and if STSTEN has been allowed. STSTACK is (CMOS) logic high when power is turned on, and goes low when STSTEN is asserted and CW is not true. 6 GATE input, CMOS/TTL, isolated by 1 k from rear panel Gate BNC. 7 STARTRTN START return

43 Chapter 6 FG-101 Pulser 43 8 STSTEN Start-stop enable, TTL/CMOS input. When STSTEN goes high, after a delay of about msec (relay switch), control of the pulse is transferred to START and STOP. When STSTEN goes low, gate control is returned to the GATE input. This requires that the device asserting STSTEN issue a stop pulse after STSTACK goes low to put the intensifier into a known off state. 9 STOPRTN STOP return.

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45 Problems and Solutions Appendix A Temperature Lock Problems Intensifier Alarm Detector doesn t achieve temperature lock If the indicator hasn t not turned green after 30 minutes, the most likely causes are as follows. If you cannot correct the problem condition entirely, try raising the Temp. setting a few degrees. The Nitrogen flow is too high. Correct flow rate ( ml/minute) can barely be felt on the lips. Insufficient coolant flow. Coolant flow should be 1-3 liters/minute. Increase the coolant flow rate. Coolant is too warm. Use cooled water or a coolant refrigerator. Too much glycol in the coolant mixture. 100% glycol is not able to carry away the heat of the Peltier as effectively as a 50/50 mixture. Detector loses temperature lock The internal temperature of the camera has gotten too high, such as might occur if the operating environment is particularly warm or if you are attempting to operate at a temperature colder than the specified limit. If this happens, an internal thermo-cutout switch will disable the cooler circuits to protect them. Turn off the cooler switch on the front of the controller. Typically, camera operation is restored in about ten minutes. Although the thermo-cutout switch will protect the camera, users are advised to power down and correct the operating conditions that caused the thermal-overload to occur. To reduce the risk of detector damage, ICCD detectors are equipped with an audible alarm in the detector head, activated when the intensity of light falling on the image intensifier exceeds a preset threshold. While the alarm is sounding the photocathode and MCP power are temporarily disabled. The detector window must be immediately covered or the controller must be turned off until the illumination level is readjusted. If the alarm sounds continuously even when the illumination level is adequately low, its threshold may have to be internally readjusted at the factory. 45

46 46 ICCD Detector manual Version 3.F Some controllers may also provide an audible alarm, indicating the same over-threshold condition as the detector alarm. Note that it is normal for the alarm to sound momentarily when the system is turned on. CAUTION Contact the factory at once if sporadic or continuous unwarranted alarms occur. They may indicate intensifier damage or another situation that requires immediate attention. Excessive Readout Noise Excessive readout noise with the intensifier off may indicate moisture accumulation on the CCD. Moisture accumulates on the CCD if dry Nitrogen is not appropriately used as described in Chapter 3. This should be corrected promptly or permanent damage will likely. Normal camera noise is a function of the gain setting and temperature as well as CCD type, but is typically in the range of 2 counts RMS, (10 counts peak-peak). This is on top of an offset which typically is about 100 counts. Moisture accumulation produces a coarser noise with many spikes >50 counts. If these types of spikes occur, especially after the camera has been in use for an extended period, turn off the system immediately. Increase the flow of Nitrogen to 2-3 liters/minute for at least 30 minutes. If excessive readout noise continues have the unit serviced by PI personnel immediately.

47 Appendix B ICCD Detector Specifications Common ICCD Specifications The ICCD detector specifications below are good for all ICCD detectors described in this manual. Specifications specific to an ICCD with a particular option, such as the FG- 101 or the CPC-100 option, are listed separately. Note that specifications are subject to change. Also, specifications may vary depending on the type of CCD and image intensifier selected. The following numbers are provided for general reference only. Operating parameters CCD Arrays: EEV full frame or frame transfer, µm pixels, MPP or non-mpp; EEV frame transfer, µm pixels, non-mpp only; EEV full frame, µm pixels, MPP only Thomson , µm pixels, MPP only Image Intensifier: 18 mm or 25 mm (LD models) Method of Coupling: 1:1 fiber optics for all except ICCD-576LD-E and ICCD-512 EFT models, which use 1.5:1 fiber optic reducers Vignetting: With fiber optic coupling there is no vignetting. ICCD-1024E detectors will show an 18 or 25 mm diameter which represents the diameter of the image intensifier. For other ICCD detectors all pixels are illuminated. Spatial Resolution: 85 µm spot size FWHM on CCD for 1:1 coupling, 60 µm spot size FWHM on CCD for 1.5:1 coupling Geometric Distortion: < 1 pixel Sensitivity: Variable gain adjustment allows sensitivities from 1-70 counts per photoelectron for detectors with 1:1 coupling, 1-35 counts per photoelectron for detectors with 1.5:1 coupling. Some phosphor choices may result in lower values. Gating Speed: Fast Gate Intensifier, 2-7 nsec FWHM; Slow Gate Intensifier, nsec FWHM; see the pulser manual for the gating limits of your pulser Gating ON/OFF Ratio: 5 x 10 6 :1 47

48 48 ICCD Detector manual Version 3.F Response Linearity: Better than 1% for the upper 95% of range in Shutter mode. Slightly non-linear response at lower 5% of range, due to phosphor nonlinearity. These response characteristics are reproducible and therefore allow complete calibration. ICCDs with better linearity are also offered, but there is an associated loss of gain. Photocathode Dark Charge (EBI): Red-blue enhanced, < 5 counts/pixel-second; Red-enhanced, < 15 counts/pixel-second; The exact dark charge in counts is of course a function of the gain of the ICCD, which varies with the type of CCD Phosphor Decay: 2 msec standard, 300 nsec phosphor is optional Spectral Range: Red-blue enhanced, nm; Red enhanced, nm Nonuniformity: Typically 12% for 18 mm intensifiers, 16% for 25 mm intensifiers CCD Cooling: down to -35 C with tap water, down to -45 C with 0 C coolant using the ST-138 Controller Photocathode Cooling: About 12 C below ambient temperature due to nitrogen flow Readout Noise: counts in Gate mode (see information for appropriate chip). Operating environment Storage Temperature: < 55 C Lab Temperature: 30 C > T > -45 C Water Flow: 1-3 liters per minute for maximum cooling Nitrogen Gas: 1-2 liters per minute initially, ml per minute during experiment Lab Humidity: < 50%. In very high humidity climates any system optics close to the image intensifier (such as a spectrometer) must also be continuously flushed with dry nitrogen gas to prevent condensation. ICCDs with the CPC-100 Option The CPC-100 option allows the photocathode of the image intensifier to be cooled significantly below room temperature. It is highly recommended for long exposures where the noise from the image intensifier becomes significant (CW mode). If you would like to add the CPC- I 00 option to an existing full frame ICCD-576 detector, contact the factory for pricing and delivery schedules. The detector must be returned to the factory for this modification. Photocathode Cooling: About 12 above the temperature of the coolant down to -20 C; Exact cooling level depends on the temperature of the coolant, as the CPC-100 itself is not thermally regulated

49 Appendix B ICCD Detector Specifications 49 Photocathode Dark Charge (EBI): Red-blue enhanced, < 1 counts/pixel-second; Red-enhanced, < 3 counts/pixel-second; The exact dark charge is a function of the detector gain, but the CPC-100 generally offers a reduction of 5 or more in EBI. Models Supported: The CPC-100 is available on the ICCD-576E, ICCD576LD-E ICCDs with the FG-101 Option The FG-101 Pulser is a combination gate pulse generator and high voltage power supply built into the detector housing of an ICCD. Because the FG-101 is a different pulser design than other, stand-alone pulsers, some of the specifications are different from the standard ICCD-Pulser configuration. The ICCD with the FG-101 option has a 9-pin connector that is not present on a standard ICCD. High voltage pulses from a standard PI pulser should never be connected to the Gate port of these detectors. Gating Speed: Fast Gate Intensifier, 5-7 nsec FWHM in Start/Stop mode; Slow Gate Intensifier, nsec FWHM; Maximum pulse width is DC Rise Time: 5 nsec Fall Time: 6 nsec Propagation Delay: 8 nsec in Start/Stop mode; 18 nsec in TTL Gate mode; Note that this is only the delay of the FG-101. If it is used with the Stanford Research DG- 535, its delay should be considered as well. Jitter: < 1 nsec in Start/Stop mode; < 2 nsec in TTL Gate mode Maximum Repetition Rate: 5 khz Pulse Magnitude: 220 V peak-to-peak Start/Stop Mode Input: 5 to 0 V falling edge, 50, compatible with DG-535 from Stanford Research TTL Gate Input: 5 V maximum input, 10 k/20 pf impedance Models Supported: The FG-101 is available on the ICCD-576, ICCD576LD, ICCD-576 Kinetics, and ICCD-512EFT

50 50 ICCD Detector manual Version 3.F Kinetics ICCDs The Kinetics ICCD camera from PI represents another of a long line of specialized intensified cameras developed in response to customer needs. CCD Arrays: Thomson full frame; EEV full frame Image Intensifier: 18 mm with fiber optic input window Method of Coupling: 1:1 fiber optics Spectral Range: nm due to fiber optic input; nm with UV-to-visible converter Aperture Adjustment: From 0 to full CCD; adjustment and alignment via precision screws Adjustment Rate: 14 pixels per turn Mask Penumbra: Approximately 2 pixels in an f/4 system Mask Edge Straightness: Within 3 µm Vertical Shift Rate: 415 khz row shift rate for EEV CCD Linearity: Depends on MCP photocurrent density; use of frame accumulation in hardware will extend the linear dynamic range; Linearity of an ICCD detector is a complex interplay of variables, including the phosphor decay time, the nature of the experiment timing, and the peak current density in the MCP relative to the MCP standing current. As a rule of thumb, pulsed experiments with time spans shorter than 1 msec become nonlinear above approximately 1,000 counts per pixel per frame.

51 Appendix C Changing the Internal Impedance of an ICCD WARNING Although all PI ICCD detectors having a Gen II Intensifier have a Gate BNC connector on the rear panel, there are three distinct types of Gate BNCs. Operating a Gen II ICCD with the Gate BNC connected to the wrong PI pulser may cause permanent damage to the ICCD detector and the pulser. Damage due to misuse is not covered by warranty. Note: For information relating to ICCDs equipped with a Gen IV Intensifier, see Operation with a Gen IV Intensifier on page 15. ICCD detectors can be gated with an external pulser or with a built in FG-101 pulser. The differences between these two models are shown in Figure 4. If you have an ICCD with an FG-101 option, never connect a high voltage pulser to the Gate BNC. For ICCD detectors without the FG-101 option, four models of high voltage pulser are available; the PG-200, the FG-100, the FG-100H, and the PG-10. The PG-200 and the PG-10 are designed to gate a detector with a 1 M termination. The FG-100 is designed to gate a detector with a 50 termination. The FG-100H, a variation of the standard FG- 100 Pulser, uses an external terminator to achieve 75 termination. A detector set up for one pulser will not function correctly with the other, and connecting a 50 - terminated ICCD to a PG-200 or a PG-10 will most likely damage the pulser. If you have purchased only a single ICCD detector with a single pulser, the termination was set correctly when the system was tested at the PI factory. If, however, you have purchased more than one ICCD, or are using an ICCD that was originally part of another system, beware! There is no way to tell at a glance how a detector is terminated. The instructions below describe testing the termination and adjusting it if necessary. Measuring the Impedance of an ICCD Detector The termination of an ICCD can be measured simply by connecting a ohmmeter across the Gate BNC on the back panel of the ICCD detector. A reading of approximately 50 indicates an ICCD set for use with an FG-100 Pulser. A reading of approximately 1 M indicates a detector set for either a PG-200 or a PG-10 Pulser. ICCD detectors for use with the FG-100H Pulser will register 1 M and must have an external terminator attached. See the section at the end of this chapter. If the termination is correct, no adjustments need to be made. Connect the High Voltage Gate Out BNC on the back of the pulser to the Gate BNC on the back panel of the ICCD 51

52 52 ICCD Detector manual Version 3.F and continue with the setup instructions. If the termination is not correct, see the instructions below. Note: The instructions in this chapter pertain to the high voltage board in the ICCD detector. This board is present in detectors shipping after June, If you find you have a different high voltage board, contact the factory for instructions on changing the impedance from 50 to 1 M. Changing from 50 to 1 M The high voltage board in the ICCD for 50 or 1 M pulsers is the same. The only change that needs to be made is the removal of a single jumper to change from 50 (FG-100) to 1 M (PG-200 or PG-10) termination. DANGER The high voltage board in the ICCD detector carries 5,000 V, enough to cause serious injury or death. Never remove the detector cover while the detector is connected to the controller. Any adjustments to the high voltage board of the ICCD can only be made by factory authorized personnel using special equipment. Turn off the controller and disconnect the ICCD detector from the controller. Wait at least 5 minutes for high voltages in the detector to discharge. Remove the lower cover of the detector, the side that has the Gate connector and the intensifier gain potentiometer. This is the high voltage board of the ICCD, part number The silkscreen of this board is shown in Figure 19. If you have a different high voltage board these instructions do not apply. Locate jumper JP2 on the board. This jumper is highlighted in Figure 19. JP2 is present for 50 operation (FG-100) and should be removed for 1 M operation (PG-200 or PG-10). Remove the jumper and replace the cover. The detector is now set for 1 M operation with a PG-200 or a PG-10 pulser. Changing from 1 M to 50 There are two possible ways to change a detector from 1 M termination to 50 termination. The first method below will work on any ICCD detector. The second method is usually only available for detectors that were originally shipped for 50 (FG- 100) operation. The easiest way to change the impedance is to use a 50 terminator from PI. One side of this adapter connects to the Gate port on the back panel of the detector, and the other side connects to the cable to the FG-100 Pulser. Do not connect the adapter to the back panel of the pulser! For pulses of the duration used by ICCDs the distance between the terminator and the ICCD electronics must be as short as possible. If you do not have a terminator from PI a standard 50, 2 W or greater coaxial terminator can be used. These terminators work on any ICCD, not just those with the board. For detectors originally shipped with 50 termination, the jumper described earlier in this chapter (JP2 on the board) can be replaced for 50 termination. If you

53 Appendix C Changing the Internal Impedance 53 purchased this ICCD detector with a PG-200 or a PG-10, you will most likely find that the two pins for the JP2 jumper were not installed. The external terminator must therefore be used. No attempt should be made at modifying the detector. Figure high voltage board ICCD Detectors for the FG-100H WARNING Do not attempt to operate an ICCD internally terminated with 50 with an FG-100H pulser, particularly at high repetition rates. Damage to the detector will likely result. The FG-100H Pulser requires the ICCD detector to be terminated with 75 instead of the standard 50 or 1 M. To avoid additional confusion, these detectors are internally 1 M terminated and use an external terminator to achieve the correct 75 impedance. Connect one side of this terminator to the Gate port on the back panel of the detector, and the other side to the 75 cable to the FG-100 Pulser. Do not connect the adapter to the back panel of the pulser! For pulses of the duration used by ICCDs the distance between the terminator and the ICCD electronics must be as short as possible. If you do not have a terminator and cable from PI, contact the factory or your local sales representative.

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55 UV Lens Appendix D Figure 20. ICCD Detector with PI f/1.2 UV Lens The PI UV accessory lens allows greatly enhanced performance in UV measurements. With an aperture of f/1.2, the PI UV accessory lens gives seven times the throughput previously available using the Nikkor f/4.5 lens. The PI UV lens is designed exclusively for use with PI ICCD-MAX or ICCD detectors having an 18 mm intensifier. The lens and detector mount in a special stand that is factory focused for optimum performance and that can be mounted on an optical bench or tripod. WARNING Handle the UV Lens - Camera Assembly by the Baseplate only! Never lift, move, support, carry or otherwise stress the Assembly using the lens, camera, or any of the other assembly parts except the baseplate. Improper handling could result in catastrophic damage which would not be covered by the Warranty. 55

56 56 ICCD Detector manual Version 3.F Both the UV lens and the camera can be easily removed from the stand or remounted on it without need for realignment. The design includes provision for field alignment should it ever be necessary. Note that the UV lens can be removed and a standard nose installed that allows the ICCD to be mounted to a spectrograph or other device for use in applications where the UV lens is not required. Although the UV lens is normally ordered with the ICCD detector with which it will be used, retrofitting to units ordered without the UV lens may be available depending on the exact configuration of the camera. If retrofitting is possible, the camera must be returned to the factory. Contact the factory for details. Specifications Spectral Transmission: 190 nm to 850 nm Type: all quartz catadioptric F-Number: f/1.2 Fraction of Unvignetted Rays: 62% due to secondary reflector masking Focal Length: 105 mm, finite conjugate lens Weight: 8.5 kg including camera and stand Dimensions: 40 x 21 x 8 cm including camera and stand Depth of Field: ±0.5 cm Focal Distance: 0.8 m to 2.5 m, optimal focus achieved at 1 m Vignetting: Less than 10% as measured from center to corner of CCD Resolution: Gen II intensifier limited Installing the Camera in the Stand A UV System comprises three major components, the Stand, the UV Lens and the Camera. The UV lens is not mounted in the stand when shipped and will have to be installed per the instructions in the next section. The camera may be shipped mounted in the stand or unmounted, depending on the policy in effect at the time of shipping. Either way, a prealignment is performed at the factory and need not be repeated in the field. If the camera is shipped unmounted, the first installation step will be to mount the camera in the stand, a fast and simple operation, performed as follows. Referring to Figure 21, a rear view of the Stand, loosen the two Clamp Screws and the Vice Jaw Screw. It is not necessary to remove these screws, just loosen them so that the Camera Clamp and Vice Jaw move freely. Carefully insert the camera into the Stand from the rear. Lift the Camera clamp so that camera can slide all the way in. There is some latitude as to how far forward the camera should be mounted. Refer to Figure 24 as a guide. If coolant fittings are installed on the camera, be sure that they remain accessible.

57 Appendix D UV Lens 57 Figure 21. The Stand Camera Clamp Clamp Screw Clamp Screw Vice Jaw Vice Jaw Thumb Screw Tighten the Clamp Screws and Vice Jaw Screw to secure the camera. Figure 22 shows the stand with the camera mounted and secured.

58 58 ICCD Detector manual Version 3.F Figure 22. Camera Mounted in Stand Camera Clamp Clamp Screw Camera Clamp Clamp Screw Vice Jaw Thumb Screw Installing the Lens Note: Earlier versions of the detector had two adjustment knobs, a large one referred to as the Macro Focus Adjustment knob (near the front end of the Carrier) and a smaller one referred to as the Micro Focus Adjustment knob (near the rear end of the Carrier). The following procedure is written for detectors that have a single Focus Adjustment knob but, wherever appropriate, the term (Macro) or (Micro) is included as part of the instructions. Depending on the shipping policy in effect at the time the UV lens is shipped, it may arrive mounted or unmounted as shown in Figure 23. Before shipping, both the camera and the lens are mounted on the stand and prealigned mechanically. The adjustments are then secured so that the camera will remain in alignment during shipping. On arrival at the customer s facility, the lens must be remounted on the stand as described in the following procedure. The lens mounting procedure follows. Carefully pick up the lens and, with the lens positioned so that the Floating Light Seal is to the back and the Carrier Engagement down, place the lens on the Carrier so that the dovetail mates. Then slide the lens back to where the teeth at the back of the Carrier Engagement begin to engage the teeth on the (Macro) Focus Adjustment at the front of the Carrier. While continuing to support the lens, rotate either (Macro) Focus Adjustment knob to draw the Carrier Engagement into the Carrier and move the lens back until it is positioned as shown in Figure 24.

59 Appendix D UV Lens 59 Figure 23. UV Lens and Mounted ICCD Figure 24. Lens Engaged and in Position to Mate the Floating Light Seal with the Fixed Light Seal

60 60 ICCD Detector manual Version 3.F Mount the Travel Stop as shown in Figure 24. It is secured to the Carrier Engagement by two screws, one on each side. When installed, it prevents the lens from being adjusted far enough forward for disengagement to occur. If the lens were to become disengaged from the Carrier, it could easily fall and be damaged. Continue to rotate the (Macro) Focus Adjustment knob to bring the lens back slowly until the Floating Light Seal slides into the Fixed Light Seal. There is considerable mechanical freedom in the Floating Light Seal assembly and it may be helpful to support it with your hand and align it as it begins to mate with the Fixed Light Seal. Note: The factory prealignment should assure that the lens will be precisely centered on the nose without need to change any of the stand adjustments. Continue the adjustment until the lens is near the center of its travel range. The setting for optimum focus will vary according to the experiment setup. Slide the Front Shield onto the UV Lens. A small slot at the edge of Front Shield mates with the Locking Pin at the top of the lens. Once the Front Shield is at the end of its travel range, simply rotate a few degrees clockwise to lock it in place. Slide the Dust Cap onto the Front Shield. The lens and camera should now appear as shown in Figure 25. Figure 25. UV Lens with Front Shield and Dust Cap

61 Appendix D UV Lens 61 Nitrogen Flushing of the Detector An ICCD camera fitted with a PI UV lens requires continuous flushing of the detector with nitrogen or dry air while operating cooled. Even if the detector is not powered it must be flushed whenever water or other coolant is circulating, as the possibility of condensation still exists. WARNING CAUTION Never use any gas other than nitrogen or dry air. Noble gases, such argon or helium, have a low breakdown voltage, which could lead to arcing that could destroy the image intensifier. If the relative humidity in your lab is high, e.g., > 40% do not operate the detector without flushing, even if you are not cooling the detector! The dryness of the nitrogen or air is critical. The colder the detector is operated, the dryer the gas must be. Also, the longer the detector is operated cold, the drier the gas must be. WARNING Be wary of unspecified dry gases intended for less demanding applications. Even a 99.9% pure air can deposit enough water to cause permanent damage to the detector. As an example, consider an ICCD camera where the CCD temperature is -35 C and where you are using air with 100 ppm of water vapor. This water vapor will be at dew point and water will condense and freeze on the CCD. Ice will continue to form for as long as this air flows over the surface of the CCD. PI recommends that gases with less than 10 ppm water vapor be used. Some examples are AIRCO compressed air grades DRY (10 ppm) Zero 1.0 (3 ppm), and AIRCO compressed nitrogen grade 4.8 (3 ppm). Equivalent gases from other quality vendors are also suitable. Connect the nitrogen source to the nitrogen inlet on the upper surface of the camera nose. Flush the detector for at least 10 minutes at a high rate (1-2 liters/minute). Then lower the rate to ml/minute. CAUTION After you have finished an experiment and have turned off the controller, maintain a gas flow of at least 2 liters/minute for at least 30 minutes. This keeps condensation from forming on the detector until it reaches room temperature. PI recommends that the ICCD detector be returned to room temperature at least one hour out of every 24 for flushing. Adjustments As previously described, a precision mechanical prealignment is performed at the factory before shipping. Other than to set the focus, it is unlikely that any adjustments will need to be reset in the field. Nevertheless, since it is at least possible that field realignment

62 62 ICCD Detector manual Version 3.F could be required, such as might be the case if a different camera were installed *, the following paragraphs describing the available adjustments are provided. Elevation As shown in Figure 26, there are two Elevation Thumb Nuts. The front one sets the elevation at the front of the camera. The back one sets the elevation at the back of the camera. Before an Elevation Thumb Nut can be adjusted, the Lock Screw that secures it must be loosened. Figure 27 identifies the rear Elevation Thumb Nut and Lock Screw. In early units there is one Lock Screw for each of the Elevation Thumb Nuts. In later units there are two Lock Screws for each Elevation Thumb Nut. Take care to tighten the Elevation Lock Screw(s) after changing an elevation setting. Figure 26. Elevation Thumb Nuts Angulation The camera, together with its support and elevation elements, rests on the Angulation Plate, which in turn rests on the Base Plate. The Angulation Plate pivots about the front shaft. The shafts are concentric with the Elevation Thumb Nuts. The angulation adjustments allow the angle of the Angulation Plate with respect to the Base Plate to be * Note that the Stand as configured for an ICCD is a little different than one configured for the ICCD-MAX. If you intend to switch from one type of camera to the other, at the very least a different Camera Clamp would be required. Contact the factory for detailed information if you need to change camera types.

63 Appendix D UV Lens 63 adjusted, and thus determine the angle of the camera with respect to the UV Lens. As shown in Figure 27, there are two Angulation Locking Screws that must be loosened before the adjustment can be made. There are also two adjustment screws, one on the left side of the camera and one on the right at the points indicated in Figure 27. To swing the Angulation Plate to the right, the right Angulation Adjustment Screw must be loosened and the adjustment made with the left Angulation Adjustment Screw. To swing the Angulation Plate to the left, the left Angulation Adjustment Screw must be loosened and the adjustment made with the right Angulation Adjustment Screw. Once the desired setting is established, the loose Angulation Adjustment Screw should be turned to where it just touches the Angulation Plate. Then the two Angulation Locking Screws should be tightened. Figure 27. Back View of ICCD and Stand Elevation Locking Screw Elevation Thumb Nut Angulation Locking Screw Angulation Adjustment Screw Angulation Plate Baseplate Angulation Locking Screw Angulation Adjustment Screw Lateral Adjustments At the front, the camera rests in a yoke, which in turn pivots inside the front vice. At the back, there is no yoke and the camera directly rests in the rear vice. As shown in Figure 28 and Figure 29, at both the front and rear there are lateral adjustments which are used to center the camera in its supports. In setting these adjustments, keep in mind that they need only touch the camera to constrain its movement and should not be tightened. Note that there is a Vice Lock Thumb Screw on one side. Loosening this screw allows the camera to be removed without changing the lateral adjustment setting.

64 64 ICCD Detector manual Version 3.F Figure 28. Left Side View Figure 29. Right Side View

65 Appendix D UV Lens 65 Filter Installation Installing an 86 mm filter in the UV lens can be accomplished easily and quickly. Each UV camera is shipped with the filter mounting components already installed. The filter itself is separate and must be mounted by the customer. Suitable filters are available from PI. CAUTION Take care not to touch the filter with your fingers or allow the optical surface to come in contact with any item which could scratch or contaminate it. We strongly advise handling the filter only while wearing powderless fingertip (or whole hand) gloves. Figure 30. Filter and Filter- Mounting Components Figure 31. Profile Drawing of Filter- Mounting Components First Spacer Filter Second Spacer Retainer O-Ring (Used only if overall Filter thickness is < 6mm.) Figure 30 and Figure 31 show the filter and filter mounting components. In addition to the filter itself, there are two spacers, a rubber O-ring, and a threaded retainer. Referring to Figure 30 and Figure 31, note that the two spacers are similar in that they are flat on one side and angled on the other. This allows the filter to be mounted at a 5 angle for optimum performance. However, as can also be seen from the figures, the two spacers are not identical in that the distance from the flat surface to the angled surface is larger for the First Spacer than the second one. The two spacers are not interchangeable and must be installed in the correct order, as shown in Figure 31.

66 66 ICCD Detector manual Version 3.F WARNING With the exception of an Allen wrench, DO NOT USE ANY TOOLS when installing or removing the filter mounting components. Be particularly careful when mounting the Retainer. It does not need to be tight. Contact pressure with the retainer set screws should be very light, just enough to keep the Second Spacer from moving about. The filter installation procedure follows. Remove the Front Shield with Dust Cap from the UV lens, which would normally remain mounted in its stand. Grasp the Retainer with your fingers and rotate it counterclockwise until it comes free. Then remove it from the UV Lens. Remove the O-ring if it has been installed. Grasp the Second Spacer and remove it from the UV Lens. Note: It isn t necessary to remove the First Spacer but, for the purposes of complete illustration, you could remove it as well. Figure 32 shows the UV System with all of the filter components removed. Figure 33 shows the UV Lens with the First Spacer (only) installed. The slope direction of the First Spacer s angled surface doesn t matter. However, these instructions assume that the First Spacer is rotated so that the widest part is at the top, as shown in Figure 31. Insert the filter so that it rests against the angled surface of the First Spacer. The First Spacer must always be installed with the flat side in, which allows the filter to rest against the angled surface, as shown in Figure 31 and Figure 34. Figure 32. UV Lens, Camera, Front Shield and Filter Components

67 Appendix D UV Lens 67 Figure 33. UV Lens with First Spacer (only) Installed First Spacer (install flat side in) Figure 34. UV Lens with First Spacer and Filter Installed Insert the Second Spacer. The angled surface goes in, against the filter as shown in Figure 31 and Figure 35.

68 68 ICCD Detector manual Version 3.F Figure 35. Second Spacer Installed If the thickness of the filter and its collar is < 6 mm, insert the rubber O-ring. If a standard PI filter is being installed, the O-ring should not be necessary. Make sure the Retainer set screws are not projecting from the back of the Retainer.

69 Figure 36. Retainer Appendix Installed D UV Lens 69 Next, engage the threads on the Retainer with those on the inner surface of the UV Lens tube. Rotate the Retainer by hand only until the back of the Retainer flange meets the face of the tube. Note: The next step will be required if the Second Spacer is moving freely after the retainer has been installed. With an Allen wrench, carefully screw in the four set screws so there is just enough contact pressure to prevent the Second Spacer from moving. This pressure should be VERY LIGHT. This completes the installation. Conversion A particularly nice feature of the UV lens is that it can be easily removed, and then the UV Lens Fixed Light Seal removed from the camera, a standard nose mounted in its place, and the camera removed from the stand so that it can be used in applications that do not require the UV Lens. This can be done without disturbing the alignment in any way. At a later time the camera can be returned to the stand and the UV Lens remounted without need to realign. The only time that realignment should be necessary is if a different camera is installed in the stand. The conversion is described in the following procedure. Removing the UV Lens Remove the Front Shield and Dust Cap from the lens. If it is in the locked position, simply rotate it a few degrees counterclockwise and pull it straight off.

70 70 ICCD Detector manual Version 3.F Remove the Lens Travel Stop (see Figure 24). The Travel Stop must be removed or installed using the mounting screws. While carefully supporting the lens, use the (Macro) Focusing Adjustment to move the lens forward until the Carrier Engagement comes free of the Carrier and the lens can be removed. The lens, preferably with its Front Shield and Dust Cap installed, should be placed in safe location. Removing the Fixed Light Seal With the Lens removed, the front of the camera will appear as shown in Figure 37. In the following steps, you will remove the four screws that secure the Fixed Light Seal, while taking care NOT to disturb any of the other screws. Then you will lift the Fixed Light Seal free of the camera and disconnect the gas tubing. Details follow. Referring to Figure 37, take careful note of the Fixed Light Seal s orientation as determined by the position of the gas connection. If the nose is reinstalled later, the same orientation must be established. Also note that the left Securing Screw is extra long, necessary because this screw provides a critical ground connection. This grounding screw must always be returned to the same position when a nose is installed. While supporting the Fixed Light Seal, remove the four screws identified in Figure 37. Again, Do not disturb any of the other screws! Set the Fixed Light Seal down as shown in Figure 38. Note that the gas tubing is still connected. Disconnect the gas tubing from the camera as indicated in Figure 39. The nose can then be removed to a safe location.

71 Appendix D UV Lens 71 Figure 37. Fixed Light Seal Securing Screws Remove this Screw (extra long grounding screw) Remove this Screw Remove this Screw Gas Connection Remove this Screw Figure 38. Fixed Light Seal Removed; Gas Tubing Still Connected

72 72 ICCD Detector manual Version 3.F Figure 39. Disconnecting the Gas Tubing Install the nose, using the four screws that previously secured the Fixed Light Seal and taking care to install the extra long grounding screw in the correct location. Figure 40 shows the camera with the Spectroscopy Nose installed. Figure 40. Spectroscopy Nose Installed

73 Appendix D UV Lens 73 Removing the Camera Viewing the Camera from the back as shown in Figure 41, loosen the two Clamp Screws and the Vice Jaw Thumb Screw. These screws should not be removed, just loosened. CAUTION Do not disturb any other screws. Loosening these three screws will allow the camera to be removed while retaining the alignment. As long as the alignment screws are undisturbed, the camera can be reinstalled at any time without need for realignment. After loosening the two Clamp Screws and the Vice Jaw Thumb Screw, work the camera free and lift it back and up to remove it from the stand. Figure 41. Removing the Camera Camera Clamp Clamp Screw Camera Clamp Clamp Screw Vice Jaw Thumb Screw Figure 42 is a grouping of the major system components handled in the course of the conversion.

74 74 ICCD Detector manual Version 3.F Figure 42. Conversion Components Grouping