Hi Issac-
Our cryostat decon/defrost procedure goes something like this:
Remove glass door to cryochamber by pulling up and gently sliding out of track.
Allow microtome and chamber to come to room temp. Wipe down exposed surfaces
of cryochamber and microtome with 1% bleach or bleach
Hi Carol,
I have used histogel for these kinds of samples and also other small,
thin tissues like insect antennae and insect GI tracts and midguts.
Since I get all my projects already fixed in whatever fixative the
investigator chooses, rinsed and placed in 70% ETOH the histogel never
Jeff, Do they give any references for the effectiveness of their proposed
method?
Tim Morken
Supervisor, Histology / IPOX
UCSF Medical Center
San Francisco, CA
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On
Jeff,
Can you give the reference for the CAP adjustment on decon for crystats
please? We have two working cryostats and average 6 to 10 cases (not
specimens) a day. This would effectively shut us down for frozens on some
days. We have third one that is currently in for repair.
I think it is strange that we are all doing similar techniques and wind up
with different outcomes using the histogel. I would be curious how many of
us are using the equipment sold with the histogel for warming and cooling
opposed to any of us who don't. we did not purchase the equipment and I
I also think that it is strange of the way Histogel processes. I have posted on
the Histonet previously about this exact problem. I worked with Jennifer
Hofecker when she was at Vanderbilt U.(sent her my Histogel and she sent me
hers) and ended up with perfectly processed Histogel blocks at our
Hi Rene,
Could you please fax me your procedure for softening nails, you said
it's the best and we do a lot of nails.
Thanks in advance.
Joyce Wilkinson
Fax # 512.9013938
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Jeff Silverman notes that CAP is moving to more rigorous cryostat
decontamination methods - mandating a weekly shutdown and wet chemical
disinfection with a tuberculocidal agent for machines used
regularly. Cryostats will have to be brought up to room temperature
for this procedure.
This weekly
Hello Rene, we also do a lot of nails. I was hoping could fax me a copy
of your protocol as well. Thank you .215-662-6539 -Fax
Albert Santiago, HT(ASCP)
Laboratory Manager
Dermatopathology
215-662-6008/6539-office
215-662-6150-fax
The information contained in this e-mail message
Hello,
We are working on an oncology project that requires the administration of
fluorescent dyes in tumor bearing mice. The dyes are allowed to circulate
and then tumors are collected, homogenized and centrifuged for fluorescent
reading. Specifically, we are looking at the dyes in the
This sounds a lot like what I did for my master's project. What I believe I
did to correct for the autofluorescence from RBCs was to have a group of
control tumor-bearing mice perfused with saline only, take readings as you
would with the dye samples, and in the analysis subtract out the saline
For many years doing IHC on mouse bones, I have used 10% EDTA, just pHed up to
where it goes into solution, with great results for frozen section
immunofluorescence. It takes 5 days. For FFPE, we use Richard Allan Decal
solution which is HCl and EDTA. Also works great and only takes 1 hour for
After cytospinning I always dry my slides thoroughly before moving on to a IHC
or IF step. I also use Superfrost Plus slides or silane coated slides to
increase adherance. I would do that then try various fixatives.
You can make a cell pellet as you suggest. Decide ahead of time if you want to
Hi Everyone,
Any tips for keeping frozen sections of cartilage from falling off the
slides during IHC staining? We should not have to do HIER, but they still fall
of the slides quite easily.
Kim
Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA
___
Hi folks,
Has anyone experienced glycogen disappearing from previously
excellent control blocks of liver. A glycogen-laden liver should be
consistent throughout should it not? (ie we can't cut through it can
we??). Or is oxidation a possible culprit here (not heard of it)? Really
interested to get
thank you all for posting about the cryostat decontamination, I have not
revised my procedure and we are due for an inspection . I think this is just
over kill, our pathologist says we have to do it if cap says. (they also made
me throw away all of the dry drys that they said were outdated?
Hi Histonetters,
Have you ever had the problem I am having now: light or totally negative
staining after service and cleaning the machine with bleach?
To explain it better:
I have immuno-autostainer from TermoFisher that was working well before I had
service from the company. After a
Air oxidation is the most likely culprit. Will specially show in sections kept
at room temperature for more than 3 weeks (as for epitope signal).
René J.
--- On Mon, 2/22/10, Greg Dobbin gvdob...@ihis.org wrote:
From: Greg Dobbin gvdob...@ihis.org
Subject: [Histonet] glycogen degeneration???
Bleach is the harshest enemy of any biological reaction and should be avoided
as much as possible or absolutely eliminated after is used, otherwise can ruin
the best staining protocol.
René J.
--- On Mon, 2/22/10, Margaryan, Naira nmargar...@childrensmemorial.org wrote:
From: Margaryan, Naira
You should aim at obtaining 4 objectives:
1- extremely thin sections (as thin as you are able to produce);
2- absolutely wrinkle free;
3- using positivey charged slides, and
4- let the sections air dry completely before starting the IHC
René J.
--- On Mon, 2/22/10, Kim Merriam
Hello! Does anyone know who the current histology lab supervisor is at Central
DuPage Hospital in IL? (the person in charge of hiring?)
Thanks!
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In addition to Rene's comments, fixation may be an issue. Deeper parts
of the block may not be as well fixed as the more superficial parts so
the farther into the block you go ... :-(
Geoff
Greg Dobbin wrote:
Hi folks,
Has anyone experienced glycogen disappearing from previously
Are there good mouse antibodies that will work on dog and cat for the following:
Factor 8
GFAP
Mycobacterium sp
S100a
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Hi Julia,
I am sorry it's taken me months to reply to this email and hopefully it's not
too late. I am only catching up on Histonet messages recently.
I stain for VE-cadherin, CD144 in murine bone routinely. RD Systems goat
anti-mouse VE-cadherin works very well in paraffin and frozen murine
Although what you say is true, it is worth noting that there are some
antigens/antibodies more amenable to acid decal. I have experienced this myself
several times recently and been burned by thinking EDTA will always give better
IHC results. At least for murine bone marrow vasculature
We have a difficulty cutting metatarsal bone . It seems that our sections are
so dried up. I was thinking that our dehydration have something to do with this
which we have placed it in a wrong processing procedure for our large bone. The
tissue is 4 mm thick and 1-2 cm in length and width and
First of all, Mollifex or any other alkaline substance will do nothing useful
to the bone.
I tend to think that you processed the bone before it was completely
decalcified and that is the cause for an incomplete infiltration and a
subsequent difficult sectioning.
René J.
--- On Mon, 2/22/10,
I have had the exact same results, one piece processing well and
another in the same cassette shrinking and hardening, when using
1.5-2% low melting point agarose (NuSieve) instead of Histogel. I was
thinking of changing to Histogel, but then this thread started last
Looking for a good system that works in recording Work Load Units.
I've inquired to CAP; they do not have any material available at this
time nor recommendations.
I've looked at CPT codes but they only reflect billable services in
pathology; descriptions are fairly general and cannot be broken
Rene-
Can you share your nail softening technique with all of us? Merci.
This e-mail may contain confidential or privileged information. If you are not
the intended recipient, please advise the sender by reply e-mail and then
delete this e-mail immediately.
The general idea is the extreme base breaks the links in keratin. I tried all
of Rene's variations and selected 20% KOH.
Well fixed nails (NBF)
Soak in 20% KOH for 30-60 minutes (checking at 10 minute intervals) until soft
and pliable. (if you let it go to long it turns into mush)
Rinse well.
Usually only if you are working with prions.
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
rick.garnh...@memorialhealthsystem.com
Sent: Monday, February 22, 2010 6:12 PM
To:
We wear gloves to section, not necessarily for safety reasons but to
eliminate squamous cells on the slides.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949
fax (303) 682-9060
www.premierlab.com
Ship to
Dear Histonetters,
The Histotechnology Group of Queensland is holding their conference in
conjunction with AIMS (Australian Institute of Medical Scientists) on the
weekend of June 11,12, 13. 2010
in Townsville, in tropical Queensland, Australia.
This is our winter, but in the tropics
Hi all,
I would like to do an immunofluorescent double labeling with two
antibodies but 1 antibody works on acetone fixed frozen tissue but not
on formalin fixed paraffin embedded tissue (CD31 BD pharmingen 553370)
and the other one works on formalin fixed paraffin embedded tissue but
not on
Although I haven't tried it myself, others have gotten CD31 from BD to work
on FFPE tissue using the tyramide amplification system on zinc buffered
formalin fixed sections. I've generally had good luck with zinc buffered
formalin myself for many antigens so it may work for your other one.
See
what is the other marker than CD31?
May you let us know?
2010-02-23
TF
发件人： Adam .
发送时间： 2010-02-23 10:55:18
收件人： Phebe Verbrugghe
抄送： histonet
主题： Re: [Histonet] Double labeling with antibodies that need differentfixatives
Although I haven't tried it myself, others have gotten
Hello Adam,
Thank you very much for this very useful information!
Do you know whether this would also work on tissue fixed with formalin
instead of zinc buffered formalin by any chance?
Also, could you give me the recipe for the zinc formalin and can I use a
standard tissue processor for
I have no idea if it would work with regular formalin. In my experience,
many antigens work as well in zinc buffered formalin without any antigen
retrieval as regular formalin with antigen retrieval. But really, you just
have to try it yourself.
On antibodies I've gotten this to work, I use
Hi Adam,
Thanks a lot, might just give it a go on just formalin fixed tissue
first.
Phebe
From: Adam . [mailto:anonwu...@gmail.com]
Sent: Tuesday, 23 February 2010 12:01 PM
To: Phebe Verbrugghe
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet]
From:Anatech Ltd. anat...@net-link.net
To:histo...@pathology.swmed.edu
Reply-To:
Date:Mon, 21 Jun 1999 12:12:19 -0400
Content-Type:text/plain; charset=us-ascii
A comprehensive review of zinc formalin up to 1992 appears in the following
paper:
RW Dapson, 1993. Fixation for the 1990's: a review
Hi Phebe
The first place to look when working up Antibodies is the Specification sheet
from the manufacturer. If you look at the BD sheet for their CD31
Immunohistochemistry is recommended for Zinc fixed paraffin sections and
acetone fixed frozeb sections but not recommended for formalin
Hi Phebe
The first place to look when working up Antibodies is the Specification sheet
from the manufacturer. If you look at the BD sheet for their CD31
Immunohistochemistry is recommended for Zinc fixed paraffin sections and
acetone fixed frozeb sections but not recommended for formalin
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