In a pinch - if you want to use the 514 line on your sorter - you can probably get away with no filter in the SSC channel and perhaps just a neutral density filter in your FSC channel, since the light scatter often is so much more intense than the fluorescence. Neal Joseph Webster wrote: > > Yes, EYFP can definitely be read on a FACScan. > The intensity is less than on the sorter with 514 excitation, > but easily detectable. > > For screening clones we run them on the FACScan, only bother > changing wavelengths on the sorter for actually sorting. > If you want to do that, you need 514 filters for FSC & SSC, > also something like 545/35 for FL1; the standard 530/30 will > allow lots of 514 laser light into the fluorescence channel. > > Joseph. > > At 02:00 24/7/2001, Nancy Harvey wrote: > >Hello, > >I have an investigator who wants to check samples that have enhanced > >YFP. The ideal excitation is 514. > >Has anyone been able to detect EYFP with the 488 excitation of > >a FACScan? > > > >I could tune the primary laser on the Vantage to 514, but have only 488/10 > >filters for FSC and SSC. > > > >Any thoughts and suggestions are appreciated. > > > >Nan Harvey > >University of MS Medical Center > >Jackson, MS 39216 -- Neal Benson University of Florida nbenson@ufl.edu
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