Removing adherent cells for Flow cytometry without trypsin - how to do it (Jun/29/2006 )

Hi-

How can I remove adherent cells for flow cytometry without trypsin? I think trypsin may be cleaving the molecule I'm staining for off the cell surface.

I was thinking of scraping them off in EDTA/PBS. does anyone have any suggestions or methods?


Thanks a lot

Mountainman

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See this thread

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Hi,
In our cases we are using scraper and by pipeting up and down gentelly you can remove these adhered cells.
But care should be taken thats they not be stredded due to vigrous scraping or pipeting.
i thinks pipeting work very well and littile chnce of contamination and mixing of cells fromother groups.
For further quiary you can contact me without any hesitation on my mail id awadh507@gmail.com
all the best
awadh

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I already tried to avoid trypsin and it works,
I didn't even scraped the cells, I just pipeted up and down (in PBS with 0.5% BSA) gently for several times and eventually they detached.
Additionally I have checked under light microscope to make sure they were not in clumps.
It worked fineand save a lot of time with trypsin and trypsin nactivation and washes...

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