RE: [Histonet] RE: IHC on tissues processed for EM

From:Tony Henwood

I agree with you.

If you don't use Gill's - why are you in the Middle Ages?

I personally have used Gill's (a commercial formulation) and was not
impressed. In bowel sections, I had the mucin staining and after only a few
days both haematoxylin and alum precipitate. Performance was definitly not
up to scratch compared to the Modified Harris's we were using at the time.

This was some 10 years ago, so maybe the commercial formulations have
improved (I hope). My recommendation is to use the techniques and stains
that suit your laboratory and meet your QC requirements. Remember we must
always be on the look-out for something better.



Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318

-----Original Message-----
From: hymclab [] 
Sent: Tuesday, 30 November 2004 8:31 AM
To: 'Gudrun Lang'; Histonetliste
Subject: RE: [Histonet] RE: IHC on tissues processed for EM

I too think that no question should be lableled as "easy".  There are no
stupid questions.  I recently went through an issue about a stain that I
have been doing for over 20 years.  Couldn't figure out what was wrong.
Technical services at two companies were stumped until someone came up with
a possible answer.  I tested it and they were correct.  Someone else on the
Histonetlist just had the same problem and I responded with what we found to
be our problem.
Then I got slammed by someone who couldn't have even read the whole message.
I was answering someone'e problem about their mucicarmine stain not working.
In my response to this person I used the terms Mayer's mucicarmine and
Weigert's Hematoxylin.  This other person that slammed accused me in living
in the middle ages for using Mayer's Hematoxlyin and Weigert's hematoxlyin
and said I should try Gill's hematoxylin.  I didn't say I used Mayer's
hematoxylin, I was referring to the name of the Mucicarmine stain.  Secondly
I had never heard of using Gill's in place of Weigert's in the Mucicarmine
stain.  I e-mailed him back I thought maybe he was confused---he said he
wasn't and he said that he'd never even done a Mucicarmine stain.  I rest my
Things like that make you want to be a lurker instead of trying to help


-----Original Message-----
From: Gudrun Lang [] 
Sent: Thursday, November 25, 2004 3:09 AM
To: Histonetliste
Subject: Re: [Histonet] RE: IHC on tissues processed for EM

I feel about saying some words about your message.
You complain, that members of the histonetlist have no basic knowledgment.
But what is basic? For me, in a routine surgery histolab, EM is very, very
special for example. I belong to those people, that work in a small corner
of the whole histological techniques. But I am interested and want to learn.
You say, people have to use databases, text books etc. In my case I have no
access through my lab to those things and they are'nt cheap (and first you
have to know where to look). I am very happy about the histonetlist. Here
are specialists, experts from every field and also "workmates". Please be
tolerant with "easy" questions and problems, perhaps the person considers
the problem more difficult than you know. friendly greetings Gudrun Lang

----- Original Message ----- 
From: "Laszlo Komuves" 
Sent: Wednesday, November 24, 2004 8:44 PM
Subject: [Histonet] RE: IHC on tissues processed for EM

There are hundreds, if not thousands of papers demonstrating successful
localization of intracellular antigens at EM or LM level using tissue
samples processed conventionally for EM (i.e. PFA/GA fixation, Os
postfixation, epoxy resin embedding).

Look  for the now classical papers by Roth and Bendayan or their early
reviews. Also the EM textbooks published in the 80's have plenty of

And let me also add a personal comment of frustration: Histonet used to be
about sharing experience and wisdom, mentoring and teaching those have were
(are they still?) eager to learn or perfect their skills. I personally
learnt/benefited from many messages posted. Now people are asking advice
about coverslip sizes? So thanks to the curse of the Internet, journal
articles and text books are used only by a dedicated few, electronic
databases (libraries, PubMed, journal archives, reagent search sites, even
Google) are not searched, and now even the vendor catalogs are not opened,
because on the Histonet somebody will respond? How will users differentiate
between solid information (based on scientific knowledge, practical
experience) or just plain ignorance? Where is critical thinking? What
happened to a once honorable guild of skilled craftsmanship, when members of
the community are  lacking elementary knowledge (just a few recent
hair-raising topics: H&E staining, pH, and the list goes on and on)?  And of
course we/they are constantly offended/complain about the lack of
recognition by our/their peers.

Feel free to get angry with me. I certainly welcome arguments and
criticism.. As for me, I go back to find something useful in one of my

Laszlo Komuves

FROM:  László G. Komuves PhD
Senior Principal Scientist,
Manager, Microscopy Core Laboratory

650 Gateway Blvd., South San Francisco, CA 94080
Phone: (650) 246-6905, Fax: (650) 624-7540

Date: Tue, 23 Nov 2004 08:02:24 -0500
From: Michele French 
Subject: [Histonet] IHC on Sections Processed for EM?
Message-ID: <>
Content-Type: text/plain; format=flowed; charset=ISO-8859-1

Good Morning Histonet! A colleague of mine did some EM work and found
something interesting. Our pathologist wanted me to try to do some IHC to
further characterize what is present in the section. Our EM person said it
would never work. I did not think it was possible either, but I thought I
would ask anyway. Has anyone tried doing an immunostain on plastic (Epon)
sections from tissue fixed and processed for EM? I am always up for a
challenge, but I am really busy right now and don't want to waste my time if
there is really no hope! Thanks in advance,  Michele French

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