Hi Alexander, I tried using the 407nm line on a Krypton laser through a 505nmSP dichroic past a 500LP Barrier filter because it was all I had, but you could sense it wasn't optimal, I think I was cutting off the gist of the emission. I am going to try a 480/20 barrier filter the next time because the emission for cyan is more like 470 - 490. I was told by the microscopy people however that under a scope you can get 488 excitation to work for cyan, so I'd bet 457 would work but suboptimally, the peak for ex is around 430 with 407 and 457 being OK but equally suboptimal. Gene Manager, Flow Cytometry Facility UCONN Health 263 Farmington Ave. Farmington,Ct. 06032 http://flowcytometry.uchc.edu -----Original Message----- From: Alexander Schmitz [mailto:axs@mb.au.dk] Sent: Monday, March 15, 2004 5:47 AM To: cyto-inbox Subject: ECFP analysis on Vantage with 457nm line (secondary laserline) Dear Flowers, On a FACSVantage SE: Is anybody using the 457 line of a Spectrum 70 laser as a secondary laser to excite ECFP ? The primary laser is 488nm. What is the best filter ? 470/15 , 485/22 Do I need other filters (eg. blocking filters ?) Which beads can be used for the laser alignment ? It would be great if you could share your experiences with me. Many thanks Alex -- ______________________________________ Dr. Alexander Schmitz FACS Core Facility Department for Molecular Biology (MBI) C.F. Moellers Allee 130/3 DK 8000 Århus Denmark axs@mb.au.dk Tel: +45 8942 2676 Tel: +45 2338 2429 Mobil Fax +45 8916 6500 ______________________________________Received on Tue Mar 16 14:18:01 2004
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