Dear Bill, Thank you very much for your reply. The PI who asked me about CFP & GFP is getting back to his collaborators re: other colour reporters for this grant proposal. If the interest heightened we would explore adding another laser as you suggested. Thanks again, and just to add that I have found your website extremely useful! Yours, Andy --On Thursday, October 24, 2002 0:23 -0400 William Telford <TelfordW@mail.nih.gov> wrote: > Hi Andrew... > > CFP "likes" a violet or blue excitation source - krypton-ion 407 or 413 > nm violet, or argon-ion 457 nm blue lines all provide > not-completely-optimal but decent excitation. Unfortunately these lines > can't be had from an Enterprise II. 488 nm won't work. > > The cheapest solution for CFP on the Vantage is to obtain a violet laser > diode from Power Technologies or Coherent and set it up in the middle > (third) laser position, where your UV beam is now. These relatively > low-power (10-20 mW) laser sources provide surprisingly good excitation > of CFP. You will need to set up some sort of mounting system - we've > used a laser post mounting system from Newport. You can set up the whole > system for a good bit less than $10,000 - much less than a water-cooled > gas laser. > > We have some sample data for violet laser diode excitation of CFP on our > website at http://home.ncifcrf.gov/ccr/flowcore/index.htm (direct access > at http://home.ncifcrf.gov/ccr/flowcore/VDL.htm). If you are interested > it trying this, we can send you more specific info. > > Good luck, > > Bill Telford > NCI-NIH > > At 12:11 PM 10/23/2002 +0100, Andrew Herman wrote: > > > Dear Flow-ers, > I have a colleague who wishes to analyze and sort cells that bear GFP > and CFP (cyan fluor. protein) reporter constructs. > Has anyone had experience doing this type of analysis with a Vantage SE > and an Enterprise II laser (6 color config). I realize that using a 488 > nm line is suboptimal for CFP, but is it feasible? > > Any info would be very much appreciated. > > Thanks, > > ---------------------- > Andrew Herman > Department of Pathology & Microbiology > University of Bristol > University Walk > Bristol BS8 1TD > Tel. +44 117 928 7511 > Fax. +44 117 928 7896 > A.Herman@bristol.ac.uk > > ---------------------- A Herman, Pathology & Microbiology A.Herman@bristol.ac.uk
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