Sarah, I fully agree with Randy Fischer about the changes in scatter properties after fixation/permeabilization. I have used BD's Lysis solution and permeabilization solution to determine IL-2. IL-4, and IFN-gamma intracellularly in cultured human T cells. I found significant reduction in the forward scatter after permeabilization. This is true in the human T cell leukemia cell line, Jurkat also. Jay Sundararajan Jayaraman, Ph.D. Research Assistant Professor Dept. of Pathology University of Miami Medical School DRI Building Room 5018 (R-134) 1450 NW 10th Avenue Miami, FL 33136 Phone: (305) 243-6100 Fax: (305) 243-4553 > -----Original Message----- > From: Fischer, Randy [SMTP:RFischer@therimmune.com] > Sent: Friday, November 19, 1999 10:20 AM > To: Cytometry Mailing List > Subject: RE: Intraprep reagent & light scatter > > > Sarah, > > A helpful point about intracellular staining is that in general all the > fix/permeabilization procedures available, either commercial or home > brew, will cause a change in the scatter properties of the cells. > Slight adjustments in your settings on the controls I know you run will > bring back your populations and this is perfectly acceptable. > > Have fun, > > Randy T. Fischer > TherImmune Research Corporation > 9700 Great Seneca Hwy > Rockville, MD 20850 > (240) 453-6256 > RFischer@therimmune.com > > > ---------- > > From: Sarah Lawson > > Sent: Saturday, November 13, 1999 5:33 AM > > To: Cytometry Mailing List > > Subject: Re:Intraprep reagent & light scatter > > > > Dear All, > > > > I am relatively new to flow cytometry and have a query regarding > > Coulter Immunotech Intraprep reagent. Has anyone got any experience > > with this product? We are finding a quite marked change in scatter > > properties following its use, although we can obtain the same scatter > > pattern by increasing FS and SS total gain. Does this matter? > > > > Any comments would be gratefully received. > > > > Kind regards, > > > > Sarah Lawson > > ( isaac.lawson@btinternet.com) > > > > > >
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