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Mark Cozens cozens at
Tue Aug 18 19:10:29 EST 1998

Dear flowland
	I am posting this message for a student in the department. He has
been experiencing trouble getting viable cultures after sorting. Sorting is
based on a two colour system. The viability has been checked (by 7AAD) at
every step of the procedure with the following results:
		% viability 1: prior to sort 			80%
			  2: post sort after 3 days culture 	1%
			  3: post sort (unlabelled) 3 day culture	1%
			  4: after 3 days culture, no sort 	70%

Cell in 2 and 3 were treated identically other than 3 being unlabelled for
the surface markers used in sorting. Cells in 4 were stained with the sort
antibodies (as a control) but not sorted.

Sort conditions;
	Sheath = ice cold PBS
	Collecting into 15ml tube containing 1ml culture media (10% FCS)
	Changing collection tube every 15 min (roughly 5ml total volume)
	Cells suspended in cold culture media

If any of this sounds familiar, or if anyone has any suggestion about
possible solutions, pleeeease email me .

Mark A Cozens
Division of Pathology
University of Tasmania
ph	03 62264828
fax	03 62264833

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