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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