Peter- I just returned from vacation so I don't know what feedback you have received regarding krypton laser use in a three laser system. We have a violet-enhanced krypton as the third laser on our modified BD bench that is interfaced to the MoFlo electronics. There are two uses for this. With GFP there are mutants which have 406nm excitation without significant 488nm excitation and mutants which have 488 excitation without significan 406nm exciation. Thus the krypton allows for two color GFP assays - Anderson, et al. PNAS Vol 93 Aug 1996, pp 8508-8511. The 406nm krypton line is also useful in immunophynotyping, allowing for efficient excitation of cascade blue. This dye, directly conjugated to various antibodies, has been in routine use in the Herzenberg lab for some time now with good results. (Cascade blue is very poorly excited by UV, but with a two step staining technique can be used for bright surface markers - old article in Cytometry on this). One drawback, of course, is cost. The Coherent Innova 302 laser is not cheap. -Marty Bigos Stanford Shared FACS Facility =========================================================================================== Hello, A customer is interested in setting up their new MoFlo with the most flexible 3-laser system for their applications. One application would be the detection of GFP. I wanted to ask if their is any advantage in the use of the wild-type GFP with a krypton laser violet excitation over the other variants? I guess the broader question is what utility is their for a krypton laser these days, especially in a 3-laser system? Thanks. Peter -- -------------------------------------------------------- Peter A. Lopez 800.822.9902 Applications Scientist 970.226.2200 Cytomation,Inc. PeterL@cytomation.com 400 E. Horsetooth Rd. Fort Collins, CO 80525 ---------------------------------------------------------
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