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General information
The system can only image green GFP/FITC, red DsRed/RFP/TexsRed and blue
DAPI dyes.
The Andor iXon DU 897 EM-CCD camera has a maximum resolution of 512 x
512 pixels. It captures black and white images with a 16-bit depth of 65,536
grey levels (with 0 being pure black- and 65,535 being pure white).
Zen Black doesn’t automatically save images – you must save them as you go
along.
Use ‘snap’ in Zen Blue to clear noise from the camera detectors before
imaging. Setting ‘snap’ for every image will slow down acquisition a lot, but
you can just ‘snap’ clear at the beginning of an image sequence. Adjust
exposure time, laser power and EM Gain for brightness in ‘Acquisition’.
Unlike the Zeiss Spinning Disk microscope in OPIC, you can’t go above 300 in
EM Gain with the camera (it’s locked in software ). The EM Gain going from 1
to 200 goes from the equivalent of ½ second to a 20ms to frame rate. Avoid
putting too much light onto the cameras EMCCD sensor. The EM-CDD camera
is 16-bit (64,000 shades of grey).
Leave the Elrya system on for 5 minutes before scanning as the Andor EM-CCD
camera needs time for its electronics to cool down.
The system has three laser lines at 405nm DAPI, 488nm GFP and 561nm
TRITC/DsRed. There are four different emission filter sets for DAPI, GFP, DsRed
and Cy5 far red, selectable via Zen Black.
In Zen Black click Live for a faster ‘preview’ imaging with more noise.
You can save configurations and the menu screen setup in Zen Black. You can
reuse microscope settings from a stored image. The images are generally saved
in Zeiss *.czi format although you can use *.lsm. To prevent PC crashes with
Zen Black, it’s a good idea to limit the number of open images.
Log-in to the PC via the account ‘
cyto\lsm user
’.
