Dear Gene, We have used sapphire and the similar gfp clone, VEX (see Anderson MT, et al., Simultaneous fluorescence-activated cell sorter analysis of two distinct transcriptional elements within a single cell using engineered green fluorescent proteins.; Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8508-11. and Anderson MT, et al.,Pairs of violet-light-excited fluorochromes for flow cytometric analysis. Cytometry. 1998 Dec 1;33(4):435-44). Our set-up here at UMMS is similar to the one described in these papers, in that we use a 407 nm laser (Coherent Laser Inc., Krypton laser). This laser is also very good for both Cascade Blue and Hoechst for DNA profiling. If you have further questions, feel free to contact me. Best wishes, 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 > ---------- > From: Pizzo@nso1.uchc.edu > Sent: Thursday, August 12, 1999 1:26 PM > To: Cytometry Mailing List > Subject: Sapphire GFP/BD LSR > > > > Colleagues, > > A researcher here at UCONN would like to use flow to > sort cells expressing the sapphire -GFP construct. > The excitation profile appears to require a 406nm > beam. I was wondering if anyone has experience with this > GFP construct and could tell me what laser they are using > to produce this line (HeCad?)? > > 2nd question, any thoughts, experiences with BD's new > LSR tabletop analyser? > > Do you think an 8mw UV beam is sufficient for basic > immunophenotyping utilizing cascade blue or Hoechst > for DNA profiling? > > Thanks for your help. > > Gene Pizzo/UCONN Health > >
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