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3.2)   Centring the Bypass Image

Once a rectangular region of interest has been defined, this image can then be centred in Bypass 
mode.
1.   

The XY controls for the Bypass image

 are located on the 

input port

 of the OptoSplit.

2.  Using the 1.5mm allen key provided, adjust each of the two controls in turn whilst observing 

your single image on the camera. One control will result in a horizontal movement, the other 
vertical.

3.  Centre the Bypass image using an iterative approach with the two controls. A rectangular Re-

gion of Interest within your imaging software is useful as a reference tool to ensure the image 
is centred and square.

4)   Obtaining a split image

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.

1.  Slide the 

Bypass / Split lever 

to ‘Split’.

2.  Install the calibration cube, by removing the magnetic 

door cover

 labelled REFLECTED PATH.

3.  Insert the calibration cube, which has a magnetic fixing and will easily locate into place, with 

the small handle facing outwards.

4.  Replace the door cover.
5.  Ensure the 

rectangular aperture

 is closed enough in both dimensions to see two distinct 

images.

4.1)   Installing the Calibration Cube

Summary of Contents for OPTOSPLIT II BYPASS

Page 1: ...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 o...

Page 2: ...OptoSplit using the two screws provided 2 Screw the support legs to the end of the bar 3 The height can be adjusted on each support leg we recommend a spirit level is used for final adjustment 2 Key...

Page 3: ...arch co uk 2 1 OptoSplit panels B Left side panel A Top Panel BYPASS SPLIT OPTOSPLIT cairn research co uk Bypass Bypass split lever XY controls for Bypass image Split control Rectangular aperture adju...

Page 4: ...d 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 chec...

Page 5: ...door cover 3 Adjust the two rectangular aperture adjusters one horizontal the other vertical to create a single rectangular region of interest visible on your camera 4 Ensure the image is square with...

Page 6: ...imaging software is useful as a reference tool to ensure the image is centred and square 4 Obtaining a split image The predominant operating mode of two identical images projected onto each half of y...

Page 7: ...nd horizontally using the provided 1 5mm allen key on the H V controls on the magnetic door cover Once you are aware which channel is which turn your attention to the transmitted image and position th...

Page 8: ...l ensure maximal field of view is projected onto each half of your camera sensor Vignetting may be present once split mode has been optimised This is characterised by one image appearing darker on one...

Page 9: ...end 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 opt...

Page 10: ...3 Remove the blank cover for the auxiliary drop in position reflected path Note the blank hold ers will have no angled thread indictor and act as a protector for this pocket 4 Insert the rotating pola...

Page 11: ...ugins folder in your root MicroManager folder on the C drive A MicroManager plugin for use with version 2 0 is also currently in development Keep an eye on our website for more details Please contact...

Page 12: ...ange your microscope cube to a position suitable for Cy5 imaging 2 The mCherry emission filter will need to be removed as it will impair the Cy5 channel For ease of removal we recommend mounting the m...

Page 13: ...f required refer to section 3 2 5 Open the rectangular aperture To return to Split conditions 6 Slide the Bypass Split lever to Split 7 Replace the Cairn filter cube 8 Return your microscope cube to a...

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