counting pombe

S L Forsburg forsburg at nospamsalk.edu
Thu Jul 3 17:29:02 EST 1997


Thomas O Westerling (westerli at cc.helsinki.fi) wrote

> Im in the process of counting cells in large number and I need to
> speed
> things up a bit.
> 
> Fixing the cells and counting in Burker cuvettes would be nice but I
> have way to many samples for that to be fast.

It's hard to know what to advise since we don't know quite what
you are doing, but in principle, you have several options.
1) coulter counting.  If you are calculating cell number increases
in a timecourse (eg, for a synchronous culture), it is easy to
fix for the coulter counter and then count them all at once.  

2) hemocytometer.  Depending on how many you need to do how often,
this can be an easy as-you-go alternative.

3) Not as accurate, but for cells of consistent size, OD at
595 can give you an estimate of cell number/ml for an exponential
culture.  

> A problem wich does not make my life any easier is that the cells
> aggregate easily. Their "hugging" is really a mess when counting.
> Sonication separates them but only temporary. Is there any chemical
> around to stop this?

I'm not sure whether you mean that your cells are sticky in
culture or when you pull an aliquot to count.  Certainly sonication
of an aliquote should be sufficient if you are just going to 
count and discard. If you mean the cells in the culture 
are sticking, then there could be several reasons.  Again,
 it's hard to know what exactly,
without further info, but the commonest reason is sexual 
agluttination  as they starve, if both mating types are present.

-- 
-susan
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S L Forsburg, PhD  forsburg at salk.edu
Molecular Biology and Virology Lab          
The Salk Institute, La Jolla CA 
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