Re: [Histonet] How to Reduce Tissue Autofluorescence

[John Kiernan via Histonet]

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/00463520275ec6a5f100/largepreview.png]
(PDF) Suppressing autofluorescence - 
ResearchGate
A 'read' is counted each time someone views a publication summary (such as the 
title, abstract, and list of authors), clicks on a figure, or views or 
downloads the full-text.
www.researchgate.net
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 
Sent: 06 February 2020 10:18
To: histonet@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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Re: [Histonet] How to Reduce Tissue Autofluorescence

[Hobbs, Carl via Histonet]

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
Howevernone are great.
Multi spectral imaging is the best way forwards butexpensive.
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 
wantif 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
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[Histonet] Fw: Job Offer in El Paso

[Pamela Walker via Histonet]

From: Pamela Walker 
Sent: Saturday, February 8, 2020 10:51 AM
To: histonet@lists.utsouthwestern.edu 
Cc: Pamela Walker 
Subject: Job Offer in El Paso

Greetings Ms. Eileen A. Allison...

I saw your recent post concerning an opening in your lab. I am sorry your 
losing a great tech but are open to recieve even greater techs. I am so excited 
your lab is getting upgrades that always a plus for good productivity in the 
lab.

My name is Pam Walker. I am currently working in Austin at a high-volume lab 
for over a year. I mainly cut surgicals, derm, biopsies, prostates, specials 
and recuts. I do embed but as needed. I would definitely be interested in the 
position and I am will to relocate if hired.
I look forward to all the information concerning this great opportunity.

Pam Walker, HT
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