Leonie, We tried this about a decade ago with Pig Islets using a FacStar+ with a Beta version of the MacSort Head. Some observations: 1)The 300u tip was not extremely difficult to use, however some Islets could reach a size that warrented going to a 400u tip. 2)With either tip its mandatory to get larger than normal size tanks. Once you get the half dozen things perfect with the fluidics you don't want to depressurise to fill the tank and start all over again. (frustrating experience here). 3)We used commercially available 50u pecan pollen to size our scattergram. Anything below 50mw we thresholded out. Even with collagenase, washes etc. there was still a fair amount of trash in the bulk prep. 4) Use of Pulse width and area combined with the pollen sizing help get us in the Ball Park. I seem to recollect that using PW helped differentiate islets from acinar cells. 5)We never found a suitable Fl. handle to absolutely identify Islets vs. other components. We had some partial success with some fluorescenated lectins. We looked at about a dozen different varieties. We actually were able to sort some islets into some petri dishes and do some functional assays. We could not get the system robust enough to do on a routine basis and the use of FCM for this was dropped. Hope this helps a little. I might be able to find some of the old instrument settings, lectin descriptions etc. but would require looking into some not very well organized boxes. Let me know Sincerely, Don Walker Celltech-Chiroscience Seattle, WA Phone: 425.489.8110 -----Original Message----- From: L.gaudry@unsw.EDU.AU [mailto:L.gaudry@unsw.EDU.AU] Sent: Sunday, August 27, 2000 5:04 PM To: cyto-inbox Subject: Islets of Langerhans Hi, Would anyone have a procedure for flow sorting of the islets of Langerhans? I have been told that they are difficult to detect on forward and side scatter because there is a lot of other tissue bits about the same size. Is there some antibody that can be used to label the islets? Thank you, Leonie Celltech-Chiroscience Seattle, WA Phone: 425.489.8110
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