Hi Ann, You should always calculate drop delay from the laser that gives you a threshold signal which is usually 488 nm giving FSC as threshold. Interception with this laser beam is when detection process starts in time and is followed by sorting decision which are both calculated in delay time. Using 2-nd laser spot, you've calculated shorter drop delay which probably worked OK, especially with larger sort envelope. Regards, Sasha On Wed, 7 Jul 1999 16:14:03 +0000 Ann Atzberger <Ann.Atzberger@EMBL-Heidelberg.de> wrote: > > Hallo Flowbies, (I hope that's not a rude word) > > 2 Questions:FACS Vantage > > 1. When sorting with dual laser does one calculate the drop delay from the > 2nd laser spot or from the first?. I've been using the 2nd and sorting > seems to be o.k (haven't checked recovery) but someone else I know > calculates from the 1st. > > 2. When doing FRET analysis should the laser with the lower wavelength > always be in position 1? > > Haven't been able to find anything regarding the above anywhere. > > > slainte > > Ann > EMBL > Heidelberg > > ---------------------- Dr Sasha Sreckovic Dept Path & Micro University of Bristol University Walk Bristol, BS8 1TD, UK Sasa.Sreckovic@bristol.ac.uk 0117-928-8606
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