[Histonet] How to Reduce Tissue Autofluorescence

[John Kiernan]

There's a very brief article (downloadable PDF) from 2002 about suppressing autofluorescence, with a few references, at https://www.researchgate.net/publication/10971457_Suppressing_autofluorescence
[https://i1.rgstatic.net/publication/10971457_Suppressing_autofluorescence/links/00463520275ec6a5f1000000/largepreview.png]<https://www.researchgate.net/publication/10971457_Suppressing_autofluorescence>
(PDF) Suppressing autofluorescence - ResearchGate<https://www.researchgate.net/publication/10971457_Suppressing_autofluorescence>
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This PDF file also has short peer-reviewed histotechnical tips on 5 other topics. Fun for all there.

For something more recent on autofluorescence, try:
Davis AS, Richter A, Becker S, Moyer JE, Sandouk A, Skinner J, Taubenberger JK (2014) Characterizing and diminishing autofluorescence in formalin-fixed paraffin-embedded human respiratory tissue. J. Histochem. Cytochem. 62: 405-423.
They compared 9 procedures and favoured 3: sodium borohydride, Sudan black B and another dye called eriochrome black T. The last-named dye is CI 14645, Mordant black 11, a monoazo  dye very briefly described on page 108 in Conn's 9th edn (1977) with the preferred name chromogen black ETOO; it's not in Conn's 10th edn (2002). Sodium borohydride reacts with aldehydes and probably reduces fixative-induced fluorescence of proteins and the native fluorescence of lipofuscins. The black dyes may work by absorbing more weakly emitted light. Sudan black B can stain lipofuscin black even in in paraffin sections.

Using “home brew” reagents is always the best way to go, because you can avoid buying simple products sold at high prices with fancy names. Avoid trying anyone's unexplained "working protocol" because annotated pieces of paper get passed along in labs and can induce well educated people to do things that are obviously wrong.

It is necessary to know the reason for each step in a lab procedure. You identify as a research assistant, so you must have a boss. Probably your boss should be online along with you, asking histonetters for advice about reducing autofluorescence.

That's quite enough from me, on 9 Feb 2020.
John Kiernan  (Anatomy, UWO, London, Canada)
= = =
________________________________
From: Arun Jyothi S.P via Histonet <histonet at lists.utsouthwestern.edu>
Sent: 06 February 2020 10:18
To: histonet at lists.utsouthwestern.edu <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] How to Reduce Tissue Autofluorescence

Dear All,

Kindly share your working protocol using  “home brew” reagents to reduce
tissue auto-fluorescence.

Thank you
Arun Jyothi S.P.
Research Assistant
Cancer Research
RGCB
Trivandrum
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