Sorting Bacteria

Barren, Phil BarrenP at MedImmune.com
Fri Apr 4 10:00:56 EST 1997


Flowers!

	Does anyone have any experience with  sorting bacteria (Streptococcus)
on a stream in air flow cytometer. I am using a 
	FACSTAR Plus. 

	What I think I need is a dye that will allow me to counter stain all
the bacteria (preferably live)  to be used in conjunction 	with a dim
surface label, and then sort on the surface labeled positives. The
counter stain would need to be dim so as 	not to interfere with the dim
surface label.

	I have tried to sort with a SSC log threshold. and this does not work.
	As soon as I vibrate the stream  I loose all low end resolution which
is where my bacteria are.

	I need to use a fluorescent trigger to threshold on, this would be less
sensitive to stream vibrations which is what I 		assume is killing me on
the log SSC threshold.

	Just a note all my Scatter parameters are on logarithmic amplification.
	


Thanks in advance for your time and energy

Philip Barren

Pb



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