Alex, We also use the 458 line for CFP, (we used to use the 413 line in our krypton laser but changed that to 568 in order to get DSred with CFP) And we also use an Innova 70C (-4, not Spectrum but except for UV probably similar). We get 300mw at 458. If you're in 488 and well-tuned you'll hit 476 (.5W),473 (.1W),466(70mw), and then 458 each time becoming a deeper blue as you very slowly adjust the vertical clock-wise. Gene Pizzo Manager, Flow Cytometry Facility UCONN Health Farmington, CT. 06032 http://flowcytometry.uchc.edu 860 679 7567 -----Original Message----- From: AlexSchmitz [mailto:axs@mb.au.dk] Sent: Thursday, March 10, 2005 10:49 AM To: cyto-inbox Subject: Finding the 458nm laser line of a Coherent innova Spectrum Laser70 on FACSVantageSE Hi FlowCytonauts, I ran into trouble finding the 458nm laser line of our Innova Spectrum70 Laser on our FACSVantage SE for ECFP analysis. Actually I can tune to 6 lines below the very strong 488nm line. (UV is probably not visible) The Innova handbook says: 70C-Spectrum: 488.0: 0.25Watt; 476.5: 0.10 472.2: ---- 465.8: ---- 457.9: 0.03 Multiline UV: 0.05 So which one is the 458 ?. The first visible one ? Unfortunately I currently have no strong ECFP expressing cell line for fishing in the dark. Are there any beads that are somewhat specific for that laserline ? I tried Sphero-Yellow bright with not much success... I am running the laser as secondary laser (488 primary, outsplitted UV as laser3). I used a 649LP filter and collected the reflected light with a special 473/19 filter (as discussed previously in that group). Any help is greatly appreciated ... /Alex -- --Received on Fri Mar 11 15:38:00 2005
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