Losing the 458 laser line> I lost the laser line a few times myself, it usually just jumped past my field of view before I could see it. It is a deep violet color at 458. I used to tune our Innova 70 for Mithramycin.I got about 300mW max as Eugene does and used 150mW for Mithramycin cell cycle analysis. Mithramycin is easy to use and is a good nuclear stain for cell cycle or as a control instead of beads. Ernest Stapleton Division Manager Immunology and Genetics Laboratories Room 1524 Health Sciences Center St. John's, Newfoundland A1B 3V6 Canada 709-777-8654 -----Original Message----- From: AlexSchmitz [mailto:axs@mb.au.dk] Sent: Thursday, March 10, 2005 12:19 PM 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 Mon Mar 14 12:18:00 2005
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