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@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@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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