Ho33342 is UV-excited, if your instrument has just 488 nm argon laser, you wont see a signal. You will need to find someone with a UV laser - some FACStar-Plus have them. It will help you to check the published protocols on this - do you have access to some ? good luck, Rachel ======================================================= Rachel M. Gerstein, Ph.D. Department of Molecular Genetics and Microbiology Graduate Program in Immunology/Virology University of Massachusetts Medical School 55 Lake Avenue North Worcester, MA 01655-0002 (508) 856-1044 (508) 856-5920 (FAX) > ---------- > From: Manoj Tadem > Sent: Wednesday, November 21, 2001 12:10 PM > To: Cytometry Mailing List > Subject: Request > > > Hi there, > I am Manoj Tadem doing PhD in United Kingdom. I am > trying to establish a protocol for staining of DNA > using Hoechst 33342. I am very new to Flow Cytometry > and am short of an expert advice regarding this > particular protocol. > Its an Epics Elite Coulter Flow cytometer. Till now I > have tried about three times, but could not establish > a reliable staining procedure. > I have few questions regarding this protocol. > > 1)I have used PMT1 and PMT2 for detecting the > flouresence of Hoechst 33342. Is it the correct way? > If not kindly suggest me the appropriate PMT's for > detection of Hoechst Dye. > > 2)Which value in the statistics, should be taken as > measure for flouresence intensity? > For instance, is the X mean or area under curve or the > %value. > > Could anyone kindly help me establishing a protocol. > For which I shall be grateful to the person. > > Thanking you, > > Your's truly, > Manoj tadem > > > __________________________________________________ > Do You Yahoo!? > Yahoo! GeoCities - quick and easy web site hosting, just $8.95/month. > http://geocities.yahoo.com/ps/info1 > >
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