Re: Trypan Blue Quenching of Autofluorescence

T. Vincent Shankey (
Tue, 21 Jan 1997 17:07:01 -0600 (CST)

On Tue, 21 Jan 1997, Wallace Lauzon wrote:

> Hello,
> I've been lurking around this E-mail list for about a month now enjoying
> the discussions and the information being exchanged.
> I have recently heard of a technique used in flow based in situ PCR
> where the autofluorescence is greatly reduced by the addition of trypan
> blue. It was just a casual reference in a talk and I was unable to find
> anything more about it from the speaker.
> I was wondering if there was anyone out there with experience in this
> technique who could provide a protocol or a good reference for one.
> >
> Wallace Lauzon

I believe that the technique of quenching nonspecific fluorescence for
FITC stained specimens was developed (and patened) by Dr. Johann Kim,
while he was working with CalTag. If trypan blue is added at the same
time as the FITC-label, it will significantly reduce background staining.
While this was developed for fluorescence based immunoassays, it works
well for cytospin or tissue specimens labelled with FITC-antibodies.
Vincent Shankey
Loyola Univ. Med. Cntr.

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