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Minimum number of cells needed for cytometry? - (Dec/11/2010 )

In order to determine transfection efficiency for GFP plasmids, I'll like to count cells using a flow cytometer but my supervisor says it will take too long to expand our cells to a sufficient quantity. Why would there be a minimum number of cells needed to run through a cytometer?

-seanspotatobusiness-

Usually FACS uses 10,000 events as a positive cut off for statistical purposes - you should be able to get this many cells out of a t-25 flask easily.

-bob1-

bob1 on Sun Dec 12 22:01:41 2010 said:


Usually FACS uses 10,000 events as a positive cut off for statistical purposes - you should be able to get this many cells out of a t-25 flask easily.


Is that a T-25 flask of adherent cells? Mine are suspended and currently occupy wells in 96-well plates (300 ul medium).

-seanspotatobusiness-

One well of a 24 well plate gives you enough cells for FACS. I don't know about suspension cell culture though... What is your cell density?
rsm

-Rsm-