Hoechst,GFP,cell cycle recent discussion

From: Ann Atzberger (ann.atzberger@EMBL-Heidelberg.de)
Date: Tue May 14 2002 - 09:25:38 EST


I take back what I said about this problem being caused by the sample.

It is definitely a problem with the alignment of the UV laser in position 3
on the MoFlo.

To correct try:
aligning as usual: run sample: if problem is still present then using the
height adjust control bring the neg cells down into the GFP neg "Zone'
Remove sample. Run UV and Green bead mixture, realign but watch the focus
ctrl! - this focus control is what causes the problem- if the beads on
focussing migrate into the GFP pos area then reposition them again with the
height adjust control not the focus ctrl.
Even slight maladjustment of the focus control cause your cells to move up
and unfortunately it is difficult to see the problem using beads.
Hope all interested understand what I'm talking about, if not let me know.




most humbly
Ann Atzberger
EMBL



This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:26:09 EST