Sorting - LS threshold?

Marty Bigos mbigos at gladstone.ucsf.edu
Mon Jun 21 10:43:24 EST 2004


Ray -

This depends upon what you are sorting for. Here is what we do at our site.

If the sort is for cells that will be grown in culture we tend to 
threshold high to eliminate counting of debris. This increases our 
sort throughput, and generally the randomly sorted debris doesn't 
affect viability.

If the sort is for DNA/RNA/protein analysis, we generally threshold 
low to exclude debris, since randomly sorted debris could be a 
contaminating artefact to the subsequent measurements.

Marty

Marty Bigos
Director, Flow Core
Gladstone Institute of Virology and Immunology
Building 3 SFGH Rm 509
415-695-3832

At 12:50 PM -0600 06/17/04, Ray Hester wrote:
>One more question about sorting - does it matter whether or not you 
>increase the forward light scatter threshold (to block out 
>dead/debris events) during the sort?  I seem to remember from 
>somewhere that it was best to leave the FS threshold low, i.e., open 
>as opposed to closed, but have forgotten the logic.
>
>Thanks again.
>
>Ray Hester
>rhester at jaguar1.usouthal.edu


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