Re: Center Stream Collection

Miller, Mark A. (mamiller@wista.wistar.upenn.edu)
Tue, 28 Jan 1997 11:09:17 -0500

Remember that the center stream will contain any particles that did not
pass the discriminator, those that did not fall within a live cell gate
according to scatter paramteres, and particles that were aborted because
they were in neighboring droplets.

Imagine staining human PBMCs with FITC-CD4 and PE-CD8. You could sort
FITC+/PE- to the left and FITC-/PE+ to the right. One might think that
collecting the center stream would be a good way to recover the B
lymphocytes (CD4-/CD8-). If all goes well, all of the B cells will be
in the center stream, but they will be in small proportion to all of the
particles/cells mentioned in the paragraph above. Furthermore, the
dilution factor of the center stream is much, much higher than that of
the side streams.