3 replies to this topic

[seanspotatobusiness]

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?

[bob1]

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.

[seanspotatobusiness]

bob1, on 12 December 2010 - 02:01 PM, 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).

[Rsm]

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