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Spinning Disk User Guide

Fast Z-stack and Multi-wavelength Acquisition

The Spinning Disk Confocal can do multi-channel Z-stack acquisition as fast as the camera can run - 30 frames per second, normally. Currently this mode does not work for DAPI and Brightfield acquisition, but we should have DAPI working by October.

To get this to work, you need to set the 'Mode' group in micro-manager to 'Fast-Confocal' and use channels in the 'Fast-Channel' group. You also need to use identical exposure times for each of your channels. If you do this you should get the maximal acquisition speed.

In general I think it should be safe to always use the 'Fast-Confocal' mode, but this has not been extensively tested. If you do use this mode the scope should always try and run as fast as it can. For instance, if you use the 'Channels' group while in 'Fast-Confocal' mode, it won't be able to do fast multi-color acquisition, because it has to move the filter wheel, but it will still do fast Z-stack acquisition if you select 'Slices first'.

When using the 'Fast-Channel' channels, there is a single multi-pass emission filter in the detection pathway. This means that the dye selectivity only comes from switching the laser line. As a result, you may see crosstalk between the channels. In particular, crosstalk between mCherry and GFP has been reported.

/var/www/html/dokuwiki/data/pages/spinning_disk_user_guide.txt · Last modified: 2011/09/02 13:15 (external edit)