Autofluorescence can be brought on by certain endogeous tissue
constituents, ie. fibronectin, lipofuscin and elastin, as well as by
fixation in aldehydes.
You don't say if your sections are fixed or not. If so, you should look at
using sodium borohydride (0.5mg/ml in PBS) for 5 minutes (glutaraldehyde)
or PBS plus a few drops of 1M glycine(formaldehyde) to block any reactive
groups. ***sodium borohydride is flammable on contact with water, and
harmful by ingestion, inhalation etc.Take adequate precautions***
Another thing to consider is reducing the section thickness, if possible,
as the intensity of autofluorescence is related to this.
You also don't mention what fluorochromes you are using. It may be
worthwhile trying a fluorochrome of a longer wavelength as there is less
likelihood of any spectral overlap with the endogenous material. As I
mentioned in an earlier posting today. we have had good results switching
to the Alexa dyes (Molecular Probes).
Regional Histology Operations Manager
Bon Secours Helath Partners Laboratories
5801 Bremo Road
Richmond, VA 23226
______________________________ Reply Separator _________________________________
Author: "Miller, Barbara" at BSHSIBTW
Date: 4/12/02 11:58 AM
I am having trouble with autofluorescence in frozen liver
anyone have a way to minimize or remove this type of
advice in re this would be appreciated.
Dept. of Gastroenterology
Roudebush VA Medical Center
<< Previous Message | Next Message >>