Counting tranfected cells carrying betagal? Ideas and suggestions?

Bart Sefton bart_sefton at QM.salk.edu
Sun Jun 16 16:27:55 EST 1996


In article <31C32D02.7140 at ere.umontreal.ca>, brukneri at ere.umontreal.ca
(Ivan Brukner) wrote:

> I am trying to quantitate numbers of cells that pick up beta-gal in 
> a transfection experiment.  I have tried to count groups of cells after 
> fixing and staining but I want better statistics... The numbers I am 
> counting could be potentially quite small (perhaps 1 in 10,000 to 100,000 
> ).  
> 
> Does anyone know of a company that cells petri dishes with gridlines 
> (smaller than the standard 1 cm X 1 cm phage plates) for counting cells 
> accurately? 
> 
> If not, how the hell are people counting there cells?  I know you can
> just make extracts of plates of cells and look at absorbance for instance 
> but that won`t help me.  Also I know about facs sorting but that is a 
> little expensive for preliminary studies... 


We use a cheap, home-made solution.  We measured the width of the field
visible with our low power objective--4 millimeters--and then marked off a
grid of squares on a thin piece of plexiglass with a sharp translucent
marker such that we could see the whole of each grid square in a field. 
We put the plexiglass on the stage and then the dish on top of it and scan
up and down the rows until we have counted the whole dish.  This will give
you the number of transfected cells per dish.  Will that do?

Bart



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