Re: Hoechst, GFP, and cell cycle post error

From: Ann Atzberger (ann.atzberger@EMBL-Heidelberg.DE)
Date: Tue Apr 23 2002 - 04:19:38 EST


>missed that bit in the original post about shutting off the UV laser.



>  The UVND filter filters out the excess signal- but doesn't explain why
> the problem occurs.

In the GFP channel I use either the 510/10 or 530/40:
I use about 50-70mw of UV and also have the BSO optics for each laser-
which is what you've quoted. (I think).
I was assuming that you are working with imaginal disc cells: using a
protocol where you trypsinise and stain at the same time from 2-4 hours at
about 20 degrees C.
0.5ug of Hoechst is used cells are filtered but not washed maybe the excess
FL comes from the solution, is that possible? Howard?


Have you used a microscope to check?

Ann



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