Hi Mustafa DsRed is about 4x brighter than m-Cherry, with around twice the excitation efficiency at 488nm (~20 vs ~10%). If you have a water-cooled argon laser on any of your sorters, you may have more luck at 514.5nm. Even with a 568nm Krypton line, i¹ve never found the cherry/tangerine Œfruity mutants¹ to be very bright either in e.coli or in eukaryotic lines. As for the MEFs, if they fit through a 70um cell strainer filter, you should be fine. Simon McCallum MRC-Hutchison Research Centre Cambridge, UK On 11/9/06 21:24, "Mustafa Özbas" <oezbasm@ethz.ch> wrote: > Dear Flowers, > > previously, I used the MoFlo in our P3 security lab of our core facility with > the 488nm laser to analyse dsRed on legionella bacteria. It was successful. > > A customer of mine is planning to perform a FACS experiment : > > Analyse a fluorescent (EGFP or Cherry (dsRED mutant, excitation around 590nm, > emission 610nm) > The main goal is to determine the number of cells showing fluorescence under a > certain set of conditions. > The next steps would follow by sorting the right population. > > The work is with Mouse embryonic fibroblasts (MEFs) which are > adherent cells. How easy is it to perform FACS in such a cell type? > Would I have to consider special preparation procedures? > > There is also an Aria or Altra available for sorting in normal conditions--B_3241187102_9325556--Received on Mon Sep 18 13:38:00 2006
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