Hi Peter, For live dead discrimination, we add 50ul of a 50ug/ml stock to about a ml of cell suspension. PI max emission is around 610nm so can be detected in either your PE, PE-TxR or PE-Cy5 channel. Generally dead cells fluoresce about 2 logs brighter than live cells. On a Calibur you can use FL2 or FL3 - compensation is more intuitive if you look at it in FL2 but in some circumstances (eg very strongly expresssing GFP cells), FL3 may be better - you can compensate (green from red) via the FL3-%FL2 line. Good luck! Derek >Hi Flowers, > >Need Help on the above topic.I tried following previous threads but still have >unanswered qs.I want to run assays to count live/dead cells with PI >as a single >dye and i use a calibur with 488 argon laser,where exactly should i expect the >signal on the FL2/3 channel? Are there any concentration issues i should >consider before running my assays? > >Peter Leposo -- *************************************************************** Derek Davies Voice: (44) 020 7269 3394 FACS Laboratory, FAX: (44) 020 7269 3479 London Research Institute, e_mail: derek.davies@cancer.org.uk Cancer Research UK mobile: 07790 604112 44 Lincolns Inn Fields, London, UK. Web Page: http://science.cancerresearchuk.org/sci/facs/ In tenebris lux ***************************************************************Received on Thu Dec 2 12:58:00 2004
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