From: Jim Freyer (freyer@lanl.gov)
Date: Mon Feb 03 2003 - 19:10:32 EST
Bill The article concerning the use of xanthan gum to keep cell aggregates in suspension is given below: Freyer JP, Fillak D, Jett JH (1989) Use of xanthan gum to suspend large particles during flow cytometric analysis and sorting. Cytometry 10: 803-806. Jim >Hello, >The best method for keeping tricky cells in suspension is Xanthum Gum. >The classic reference is by Jim Jett et al, in an old issue of >Cytometry...Jim?? >What is nice about xanthum gum is that it viscous when sitting, but >fluid in motion such as through the uptake tube. >We use it to keep pancreatic islets from sinking and adipocytes from floating. >-Bill > >********************************** >William C. Hyun, Director >Laboratory for Cell Analysis >UCSF Comprehensive Cancer Center > > > > > -----Original Message----- >> From: Lidija Bosnjak >>[<mailto:lidija_bosnjak@wmi.usyd.edu.au>mailto:lidija_bosnjak@wmi.usyd.edu.au] >> Sent: Thursday, January 30, 2003 2:00 PM >> To: Cytometry Mailing List >> Subject: Re: keeping cells in suspension >> >> >> >> You can try adding 1-5 mM EDTA to prevent cell clumping and >> sticking. If the cells are >> sticking to the "standard" polystyrene tubes, another option >> may be to aliquot cells >> into a polypropylene tube (to which cells shouldn't stick) >> that will fit inside the >> standard FACS tube and analyse the cells that way. Good luck! >> >> Lidija >> >> "McCord, Robert" wrote: >> >> > Hi all, >> > >> > I have a user with HUH-7 cells that needs to keep them in >> suspension and break the >> > clumps. She is treating her flasks with trypsin and then >> adding PBS with FCS and >> > analyzing on a flow cytometer. >> > >> > She loses a lot of cells before she gets a chance to >> analyze them on the cytometer. >> > They start sticking to the tubing immediately. >> > >> > Is there a trick to keeping most of these cells in >> suspension? Any advice is >> > greatly appreciated. >> > >> > Many thanks, >> > >> > Robert McCord >> > >> > MAXYGEN >> > 515 Galveston Drive >> > Redwood City, CA 94063 >> > Phone: (650) 298-5852 >> > Fax: (650) 298-5449 >> > Main: (650) 298-5300 >> > <http://www.maxygen.com/>http://www.maxygen.com/ >> -- ---------------------------------------------- James P. Freyer, Ph.D. Technical Staff Member Bioscience Division Mail Stop E535 Los Alamos National Laboratory Los Alamos, NM 87545 505-667-8229 (office phone) 505-665-4637 (fax) ---------------------------------------------
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