[Histonet] How to Reduce Tissue Autofluorescence

[Hobbs, Carl]

Hi
Do you refer to FIF?
Or...autofluorescence?
Different...as you prob know so, apologies in advance.
The Wright Cell imaging Facility ( Toronto western research Inst) pdf: very informative
FIF: I have tried Glycine, Ammonium chloride and Na-borohyd.
I got best results with Glycine
However....none are great.
Multi spectral imaging is the best way forwards but....expensive.
Sure, in Lambda mode using Zeiss Zen on a confocal gives good results ( tho I have forgotten how to do it, sigh)
After setting up you hit the fluorescence you don't want...hit the one you want....if the wavelengths are different, what you don't want you eliminate, electronically.
Vectorlabs sell an excellent kit for FIF elimination...sure over-expensive, given the ingredients.
We found that dilution of their working reagent by x2- x4 gave best results
As good as multispectral imaging, imho.
Autofl of lipofuscin is supressed using Sudan BlackB.

Good luck




Carl Hobbs FIBMS
Histology and Imaging Manager
Wolfson CARD
Guys Campus, London Bridge 
Kings College London
London
SE1 1UL
 

020 7848 6813