sorting large particles

Nealley, Eric W Mr USAMRICD Eric.Nealley at
Wed Jul 11 09:11:03 EST 2001

Our group has been tasked to sort large diameter beads (180 um) which have
gone through a process to attach peptides.  We suggested that a new library
of smaller peptide labeled beads be used, but for now the group wants to try
and make use of the larger beads until a new library can be created using
small beads.  We have a FACStar Plus equipped with a 400 um nozzle (with
turbo sort option) and we have tried to align/standardize/calibrate with
little success. We have followed the BD macrosort procedure (lower sheath
psi and lower ddf) but again with little success. The only bead that I
currently have for alignment are flow check beads which are ~10 um (I assume
at this point that 10 um beads are not the correct ones to use with the
large nozzle).  I have ordered 100 um beads from FCSC.

-  Has anyone sorted using a 400 um nozzle?
-  Has anyone successfully sorted ~180 um beads before?
-  How do we align/calibrate/standardize the instrument?

Thank you.

Eric Nealley
Biologist, Pharmacology Division
Aberdeen Proving Ground, Maryland

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