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Cell cycle and Hoescht Stain
I have a question:
I am trying to optimise a protocol to analyse the cell cycle in some melanoma cells after treatment.
I am working with a BD LSRII (brand new) so I started to see how the machine was working.
After a little work with the software i started to obtain the first results.
I am staining my cells with hoescht , without fixation and permeabilization.
I am using 1 ug/ml of stain, and i wait 30’ before the analisys.
Now, the problem is that suddenly i cannot reproduce what i was obtaining, with any of the cell lines I was using!!!I tryed two different hoescht staining with different ability to permeate the cell, but nothing...
I do not obtain the classical cell cycle pattern anymore but just one huge peak at low fluorescence.
May I ask if anybody has some suggestion to give me?
I cannot fix and permeabilise the cells, and they are in complete medium…
Last edited by admin; 07-06-2009 at 05:00 AM.
|cell , cell cycle hoescht lsrii , cycle , hoescht , stain|
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|cell cycle||apelge||Flow Cytometry Forum||0||07-05-2009 07:07 PM|