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OPTOSPLIT II
BYPASS
This guide details initial set up and installation of your OptoSplit II Bypass (BP) image splitter. Each unit is serial numbered,
calibrated and QC'd prior to delivery, therefore minimal setup is required providing you are familiar with the key controls.
For the purpose of this manual, all words in
A supplementary Troubleshooting Guide is also available to address frequently asked questions during OptoSplit operation.
Please do not hesitate to contact us if you have any questions or issues during installation or operation
(tech@cairn-research.co.uk).
1) Mounting to your microscope
It is best practice to firstly ensure you have optimised your microscope light source in the absence of the
OptoSplit, with the camera mounted directly on the c-mount. Many technical queries are raised relating to
uneven illumination, which is often not introduced by the image splitter. Once both your transmitted and
fluorescent light source have been correctly aligned and focussed, the OptoSplit can be mounted on a 1x
c-mount; for deviations from this, please contact us for advice.
1.1) Correct camera orientation
output port
The OptoSplit
knurled section on the output port can be rotated to secure the camera, whilst also ensuring any cables
connected do not get tangled.
input port
The OptoSplit
Output port
Camera mounting thread
For correct alignment of the two emission channels, the orientation of the camera once attached to the
OptoSplit II BP unit is important. Always ensure the top of your camera lies parallel with the top of the
OptoSplit. If fitting onto an upright microscope, this camera orientation remains the same.
bold blue
refer to parts labelled in diagrams throughout this document.
connects to your camera c-mount via a
then connects to your microscope c-mount.
OPTOSPLIT
cairn-research.co.uk
email: sales@cairn-research.co.uk tech@cairn-research.co.uk
+44(0)1795 590140 www.cairn-research.co.uk
camera mounting
Input port
Bypass
M a n u a l
thread. The silver
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Summary of Contents for Cairn OPTOSPLIT II BYPASS
- Page 1 BYPASS This guide details initial set up and installation of your OptoSplit II Bypass (BP) image splitter. Each unit is serial numbered, calibrated and QC’d prior to delivery, therefore minimal setup is required providing you are familiar with the key controls.
-
Page 2: Support Legs
Support leg bar Trim adjustment and clamp screw Support legs 2) Key controls This section details the key controls for all operating modes – all labels are referred to in subsequent text in bold blue font. email: sales@cairn-research.co.uk tech@cairn-research.co.uk +44(0)1795 590140 www.cairn-research.co.uk... - Page 3 Bypass / split lever XY controls for Bypass image OPTOSPLIT Bypass cairn-research.co.uk Rectangular aperture adjusters Split control B: Left side panel Reflected H & V controls REFLECTED PATH SPLIT Split control Support leg Door cover email: sales@cairn-research.co.uk tech@cairn-research.co.uk +44(0)1795 590140 www.cairn-research.co.uk...
- Page 4 Support legs 3) Centralising the Bypass image The OptoSplit II Bypass unit requires a small adjustment once installed to ensure the Bypass image is projected onto the centre of the camera sensor. It’s best practise to ensure this is carried out prior to performing dual channel imaging, but this adjustment can be checked periodically.
- Page 5 Bypass / Split Lever 1. Slide the to ‘Bypass’ door cover 2. Ensure there is no Cairn filter cube installed in the unit by removing the magnetic labelled REFLECTED PATH. Replace the door cover. rectangular aperture adjusters 3. Adjust the two (one horizontal, the other vertical) to create a single rectangular region of interest visible on your camera.
- Page 6 The predominant operating mode of two identical images projected onto each half of your camera sensor is achieved by installation of a Cairn filter cube. For initial setup, we recommend using the supplied Calibration Cube (with a 50% / 50% mirror) and a focussed image of a graticule with transmitted light.
- Page 7 1. Turn the edge of the camera sensor. Split control clockwise Transmitted V control 2. Adjust the (using the 1.5mm allen key provided) to optimise the vertical position of this (red) image. Transmitted V control email: sales@cairn-research.co.uk tech@cairn-research.co.uk +44(0)1795 590140 www.cairn-research.co.uk...
- Page 8 TRIM adjustment 3. Slide the in one direction (diagonally) until the vignetting is removed. When moved too far, vignetting will begin to appear on the opposite image. email: sales@cairn-research.co.uk tech@cairn-research.co.uk +44(0)1795 590140 www.cairn-research.co.uk...
- Page 9 As a result, we only recommend the Chroma Ultra-Flat (UF-2) 2mm thick dichroic mirror range. The Cairn Filter Cube accepts dichroic mirrors of the following size: 26mm x 38mm x 2mm. Other dichroic mirrors will fit, but may not be optimal.
- Page 10 5.2) Polarisation (anisotropy) studies For polarisation experiments, a Cairn filter cube with an integrated polarising beamsplitter cube* is also available as an additional accessory. In addition, a rotating 25mm polariser is available to act as a ‘clean-up’ for the reflected S wave.
- Page 11 (shorter) wavelength, resulting in a single, central image projected onto the camera sensor. In Bypass mode with the Cairn filter cube in place, the single image observed will be the trans- mitted (longer) wavelength channel. Depending upon your experimental requirement in Bypass mode, there are several scenarios for optimal operation.
- Page 12 7.2) Obtaining a full field image of a longer emission wavelength than your split conditions e.g. A full field Cy5 image when splitting GFP / mCherry 1. Leave the Cairn filter cube in place and change your microscope cube to a position suitable for Cy5 imaging.
- Page 13 Close the to ensure only half the camera chip is illuminated. Although designed to generate minimal displacement when replacing the Cairn filter cube, final adjustment of split conditions may be necessary – please refer to section 4.2 Please refer to the supplementary Troubleshooting Guide for additional information not de- scribed in this manual.