User guide for rack mounted Optoscan - CAIRN RESEARCH - #19

Installation Guide

The Cairn emission detection hardware is designed for connection to the C-mount port on the side of the microscope. The dichroic mirrors and lens positions are pre-configured at Cairn and the components will normally be supplied fully assembled for location directly onto the C-mount. After locating the detection components, the appropriate cabling should be made. The photomultipliers have two connections to be made to the control modules. The HV BNC should be connected to the PMT supply module using the HV lead supplied, and the output BNC connected to the PMT amplifier input using a standard BNC lead. If additional photomultiplier tubes are present, they must be connected to separate PMT supply and amplifier modules. The acquired signals from wavelength positions 1-8 will be available at positions 1-8 on the output module by default, and connections to external data acquisition and recording systems should be made here. If supplied, the CCD camera will need to be connected to a DC power supply (supplied) and its output connected to the monitor using a BNC cable and adapter (supplied).

The infrared viewing system allows the experiment to be monitored visually both prior to and during experiments. It is an important supplement to viewing through the microscope eyepieces as it allows the experimenter to see the precise region of interest being monitored by the photodetectors, without interfering with the fluorescence signals. It incorporates several of the items already detailed, but is best tested in its entirety. Correct operation should be verified, with the photomultiplier(s) switched off or the cabling disconnected, as follows:

1. Connect the full detection assembly to the microscope with the CCD camera and monitor connected and switched on.

2. Install the 715nm or 780nm long pass infrared illumination filter in the microscope, but position it so that it is NOT in the light path at this stage.

N.B. The filter should fit into either a sliding or rotating assembly in the transmitted light path so that it can be easily removed when viewing through the eyepieces.

3. Focus a typical test sample (e.g. some unloaded cells) using the eyepieces.

1. Insert the infrared filter and divert the light path to the photometric output port using the appropriate lever or dial on the microscope.

2. Adjust the illumination level of the microscope light source until the diaphragm blades and the image of the test object are clearly visible on the monitor.

3. The microscope should require only minimal readjustment to attain best focus on the CCD camera. Adjustment of the microscope C-mount will allow matching of the image planes seen through the side port and eyepieces. If significant refocusing is needed when switching between the side port and the eyepieces, or if the object and diaphragm are not parfocal then please consult with us.

4. The field of view on the monitor will probably be more restricted than that viewed through the eyepieces. If however a suitably sized object cannot be "framed" properly on the monitor then please contact us to determine which accessory optics are required.

5. Verify the opening and closing, rotation, and lateral movement of the rectangular diaphragm. If the blades are dirty the diaphragm should be removed, allowing the blades to be cleaned using a pressurised air jet or a matchstick. The diaphragm can be removed from its holder by releasing the two knurled grub screws normally used for rotation, then sliding it out of the holder. There may be slight imperfections on the edges of the diaphragm blades, even after cleaning. This is normal and will not affect its performance.

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