Andreas, What you suggest will probably work but what might be easier is using PKH-26 (red) for one cell line and PKH-2 (green) for the other. Both cells will go through the same processing and it is easier on the cells that H-33342 staining, you also exclude the need for the UV laser. Brian Newsom Center for Cell and Gene Therapy Baylor College of Medicine ---------- From: Andreas Simm Sent: Friday, January 22, 1999 2:01 AM To: Cytometry Mailing List Subject: membrane-labeling Hello flowers, I repeat my question as I did not get the original mail from Wednesday and maybe, this mail was lost. Excuse me, if you get this mail a second time! A colleague want to make cell fusions. For this purposes, the membane of one cell population (B-cells) should be labelled for later analysis of the fusion by flow cytometry. I proposed pkH-26 from Sigma for membrane labeling, and in addition, the labeling of the DNA of the second cell population with Hoechst 33342. Cells from positive fusions should have blue nuclei and red membranes (suitable for the analysis by flow cytometry). Is this choice acceptable or does anybody knows better alternatives? We would be happy for any references. Thanks Andreas PD Dr. Andreas Simm Institut für klinische Biochemie und Pathobiochemie Versbacher Str. 5 97078 Würzburg Tel.: 0931-201-3479 FAX.: 0931-201-3153
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