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Roger Phillips <[log in to unmask]> writes:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear Jim,
> What wavelength were you using?  I recently tried to use our Zeiss LSM510
> meta NLO to image DAPI-stained fixed cells with wavelengths 750-850nm while
> also recording 1P confocal images of GFP and Cy3.  This was unsuccessful
> because the MP bleached the Cy3 rapidly and the GFP more slowly.


For fixed cell application like this you can easily sequentially
full-frame image, CY3, GFP then DAPI which will allow you to optimise
your signal:noise in each channel without undue bleaching.
Unfortunately in live cells, especially when doing time lapse these
problems are more difficult to overcome!

Ian