[Histonet] Autofluorescence and literature for getting rid of the problem

[gayle callis]

Richard, 

 

You will find what you need in a free pdf on autofluorescence from Wright
Imaging Facility in Toronto Canada.  They have a website, then download
Autofluorescence:  Causes and Cures.  Also, if you can't get rid of
autofluorescence , use a near infrared fluorophore e.g. Alexa 750.  There is
no autofluorescence seen in the NIR range.  Just Google the title and it
will come up instantly.  They also have a pdf on Mounting Media for
fluorescence work.

 

 

For the little FFPE fluorescent work we do, you can try 100 to 300 mM
glycine in TRIS buffer pH 7.4 for 20 to 30 minutes before embarking on
immunostaining.   Glycine binds free aldehydes to reduce the
autofluorescence but doesn't always work 100%.  It may reduce the problem
but not totally eliminate it.  

 

There are other references on getting rid of autofluorescence, one for GFP,
a review, but it applies to FFPE tissue even not containing GFP.   I will be
happy to  send the pdf if you wish.   

 

Gayle M. Callis

HTL/HT/MT(ASCP) 

Bozeman MT