sorting large particles
sw11527 at glaxowellcome.com
Thu Jul 12 18:39:30 EST 2001
I have used the 400u nozzle for similar efforts.
The largest bead I ran was +/-130u.
As Glenn suggested, I think that you may have other issues besides large
beads and large drops.
My guess is that your 180's sink like rocks.
I would be happy to speak on the phone with you about how I did the set-up
for the large nozzles and low pressures.
sw11527 at gsk.com
High Throughput Biology
* Tel. 919-483-3078
GlaxoSmithKline R&D Page 919-857-7768
5 Moore Dr. Fax 919-483-0585
RTP, NC 27709
> -----Original Message-----
> From: Nealley, Eric W Mr USAMRICD [SMTP:Eric.Nealley at apg.amedd.army.mil]
> Sent: Wednesday, July 11, 2001 10:11 AM
> To: Cytometry Mailing List
> Subject: sorting large particles
> 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
> of smaller peptide labeled beads be used, but for now the group wants to
> 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
> 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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