Re: Signal detection and sensitivity

From: Jens Fleischer (jfleischer@knuut.de)
Date: Wed May 31 2000 - 12:48:22 EST


Robert Adair McGilp wrote:

> I have an investigator who has been using a FACSort to analyse his FITC/
> PE/Cychrome labelled samples and gets good resolution of the populations
> of interest (dual positives). He feels that we have been unable to
> reproduce the results on our Vantage SE. We have moved the z axis so that
> the nozzle is a good inch above the laser intercept and this gives a good
> approximation of the results he has seen on the analyser.

Be sure to insert the appropriate filter in front of the FL3 detector.
Pe-Cy5 has its emission maximum at 675 nm. BD has a 675/40 bandpass
filter and a 650 longpass. Both should give a good resolution. In the
standard configuration you often find a 630/22 or 610/20 designed for
propidium iodide in front of FL3, which will not give good intensities
for PE-Cy5.

Jens



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