Re: Sorting Question

From: Daryl Webb (dwebb@waite.adelaide.edu.au)
Date: Fri Jun 20 1997 - 00:24:38 EST


Vincent Falco laments:
> 
> 
> My problem is this:During some sorts I get ,what I will call a spurious extra 
> drop. To explain more,there are times I sort to polycarb filters.When all is 
> well I get a single spot which contains the desired cell population.When all 
> is supposed not well I get my desired single spot with cells and just about a 
> quarter of inch from the desired spot I observe a spurious spot,which does 
> not contain the desired cells.


Hi, I've gottta have a stab at this one...  Its a few years since I ran a
sorter in my pevious life, but this sounds to me like a case of uncharged
"satellite drops"  merging with one of your charged drops.  Someone will
tell me if I'm off on a tangent :-)

I'm guessing 3 drop sorts here.  When the drops breakoff the joining neck 
forms a small "satellite droplet" , which depending upon phasing and 
frequency tends to merge either with the cell above or below it.

(this is from deep memory, but there is a good discussion in Flow Cyt and 
cell sorting 2nd ed,... The Yellow one...  by Myron Melamid (sp) and others)

You can see all of this through the observation microscope, have a look 
to see which cell the satellite merges with when all is well, and I'll 
bet it goes the other way when you see the "doublet".  I think you have 
an uncharged droplet merging with a charged cell, reducing the effective 
charge, causing less deflection and the appearance of the second spot.

I think its a pressure/frequency thing, but check in the old yellow bible 
as its well covered.

Best of luck
-- 

  Daryl Webb   (dwebb@waite.adelaide.edu.au)
  Dept. of Plant Science,    Waite Institute
  University of Adelaide,  Glen Osmond S.A. 5064
  Australia. Voice:61_8 8303 7426  Fax:61_8 8303 7102



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