Richard, Those of us who run core facilities face this problem quite frequently. And the best part is that she will insist on doing it, and then go away saying that a) you don't know what you're doing or b) flow doesn't work. "Sometimes you just wanna smack em!" I would try to refuse to run them, but that depends on your political situation. Figure 4 in Dr. Haugland's chapter in Methods in Cell Biology V42, PtB, (page 647) suggests that you might get a bit of a signal, but I would imagine it would take a highly expressed target. I would show her that figure and do your best to explain the limitations of this approach. And either do it for free, or make it clear that you will charge her no matter what (depending on your budget and her attitude). There's no getting around the physics of fluorescence. (Maybe we need an international organization dedicated to an increased understanding of fluorecence--we could call it the Rainbow Coalition.) Sorry, I'm buggy from proofreading! ;-) +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Steve G. Hilliard (706) 542-9474 University of Georgia Cell Analysis Facility flowman@uga.edu URL: http://floweb.cb.uga.edu/ On Tue, 2 Nov 1999, Jaramillo, Richard shared the following... > optimal excitation wavelength for Texas Red as I have discovered is 590nm. > I have discussed this with her, but she insists on analysis at 514nm. I > cannot envision getting any suitable results. Does anyone out there have > any experience with Texas Red that could shed any light on this problem? > > Thank You > > Richard J. Jaramillo > LRRI > Albuquerque,NM > Ph(505)845-1217 Fax(505)845-1198 > rjaramil@lrri.org >
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