sorting large particles

Witherspoon, Sam sw11527 at glaxowellcome.com
Thu Jul 12 18:39:30 EST 2001


Hi Eric,
	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.

	Cheers,
		Sam
Sam Witherspoon
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
> 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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