I'm using DAB Plus chromogen from LabVision and counterstained with Harris
Hematoxylin from Sigma .Then mounted using Cytoseal XYL from
Richard-Allan Scientific.I'm dehydrated the slide through graded of alcohol
then cleared in xylene prior to mounting. After done the IHC I've stored the
slide in
What mounting media are you using? Are you dehydrating? How are you storing the
slides? What DAB are you using (ie: source)?
-Original Message-
From: shazana hilda
Date: Tue, 03 Feb 2009 19:23:16
To: McMahon, Loralee A; histonet
forum
Subject: Re: [Histonet] query regarding IHC slide
Dear Loralee,
Unfortunately I'm using DAB and yet still faded. This is the first time I'm
facing this problem.
Thank You.
Shazana Hilda ShamsuddinUniversiti Sains Malaysia
Off. no: +609- 766 4440
Fax no: +609- 765 3370
Email: hilda_1...@yahoo.com
From: "M
I'm using DAB chromogen from LabVision and counterstained with Harris
Hematoxylin from Sigma .Then mounted using Cytoseal XYL from Richard-Allan
Scientific.
Thank You.
Shazana Hilda Shamsuddin
MSc.of Molecular Pathology,
Dept. of Pathology,
School of Medical Sciences,
Universiti Sains Malaysi
Does anyone use the Leica Thermal Block? How does it work? Do you
need to prewarm it? I was told to leave it in the cryostat but that
doesn't work as it gets as cold as the chucks ... ?
Thanks,
Andrea
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You have 2 options:
1-put it in the tissue processor, in the cleaning cycle, and again process it,
or
2-melt the paraffin away and press the tissue between 2 pieces of absorbent
paper, reembed it and cut it again.
René J.
--- On Tue, 2/3/09, kristen arvidson wrote:
From: kristen arvidson
You have 2 options:
1-put it in the tissue processor, in the cleaning cycle, and again process it,
or
2-melt the paraffin away and press the tissue between 2 pieces of absorbent
paper, reembed it and cut it again.
René J.
--- On Tue, 2/3/09, kristen arvidson wrote:
From: kristen arvidson
Sub
Possibly you have allowed the slides to air dry too long before
staining.
After cutting and allowing to dry for 10 minutes, store them at -70oC
prior to staining might solve the problem
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612
Hi Scott,
"slowly freezing on dry ice" may be your problem. In my honest opinion, you
need to freeze faster. Try a different method, such as cooled isopentane,
either with LN2 (coolest method) or dry ice. You should get better results
when you freeze them faster or colder.
Good luck,
Jo Dee
~~
Hi everyone... been having some trouble lately with my
cryopreservation. Tissue looks grossly fine before embedding in OCT but
then when I cut sections on the crystat the architecture is totally
disrupted with a lot of space in between cells. It almost looks like
the cells have become so shrunken a
I have never tried that type of double staining, but DAPI also stains DNA in
the minor groove and may not interfere with your Giemsa stain. You can get
DAPI already diluted in mounting media - Vector Labs makes a nice aqueous mount
one that hardens so you don't have to use fingernail polish to
Kristen
I remember reading an article maybe in the Histology Journal awhile ago. In
there it recommended rolling and blotting the tissue with a paper towel to
remove excess paraffin and then placing the specimen in a cassette and
putting that cassette directly into formalin. I never had an issues
Hi Histonians,
I have experienced some difficulties in doing snapped freeze muscle biopsies
that need to be stained with the Modified Gomori's Trichrome by Engel, W.K.,
and Cunningham 1963.
When I stained the frozen tissue that has not been post or pre fixed, I am
getting red areas within this
NO,NO,NO, CAP does not call "grossing tissue", "processing tissue". CAP
has made two categories or specimens- Processing and grossing.
ANP.11600
1. Processing is defined as a tissue examination limited to description,
inking and cutting of the specimen (if applicable), and submission of
the entire
Hi Jennifer,
I always submerge my cassettes in molten paraffin until I embed them, so
that isn't the problem. The histogel comes out of the processor hardened
and whitish in appearance. I also melted the histogel tubes in hot water
while preembedding the embryos, so that probably isn't the proble
All,
with Histogel "drying","hardening", and the loss of specimens.
I am particularly interested in artifacts of processing with algenates, among
which is Histogel. As I am analyizing the problem for our own use I have
determined that there are variables introduced with the following:
1. Age o
Hi,
Do either of you "dry embed" your Histogel blocks? By this I mean: Are you
submerging the blocks in molten paraffin prior to embedding? Are they coming
off the processor looking different, or are you finding it after you embed the
gel blocks? Does it seem that the gel square is hard and nev
What mounting media are you using? Are the slides left to dry near a UV light
from a hood or cryostat?
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
histonet-requ...@lists.utsouthwestern.edu
Sent: Tuesd
ANP.11665 refers to a non PA or non pathologist grossing tissue. CAP calls
grossing tissue "processing specimens". There has to be a manual with all
grossing techniques such as "Ackermans' Pathology" or a manual made at the
site.
ANP.22990 deals with performing FISH & ISH, there should be tech sam
Hello,
What are people doing these days when they have to reprocess fatty tissue?
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Hi all,
I was wondering if anyone was familiar with the protocol used by
manufacturers such as Menzel-Glaser regarding placing a permanent
positive charge on their glass slides. In the case of M-G they are
called Superfrost PLUS or ULTRA PLUS slides. The reason I am asking is
because I wish t
Hello,
I'm thinking of counterstaining some DAB-stained sections with DAPI. Does
anyone know if DAPI will still bind with DAB stained cells and fluoresce with
permount?
Thanks,
Reza
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Hello all. I am excited to be a new participant of the Histonet list serve. I
hope that you can provide some assistance in a little problem I am
having.Simply, I wish to photograph blood smear slides (methanol fixed) with
both Hoescht and Giemsa. I want to find items via normal light microsc
Dear Pam,
I've had the same problem. I called Richard-Allen just after they were
"absorbed" by Thermo Fisher and got no answers. They had never heard of
such a problem and didn't know how to solve it. I have heard from another
user that had the same exact problem. I lost precious samples, E6.5
Does anyone have experience with InsituPro?
http://www.intavis.com/en/In_Situ_Detection/index.php
Tora
YourBiomed.Com wrote:
Jean,
I work at IMEB, Inc in San Marcos, CA.
I'm very familiar with the instruments listed in the responses to your question.
What it comes down to is cost, closed syste
Leslie
I contacted our collegues in Germany and was given the below nformation on
the inner demensions of the Miles adapter for the CM1850 cryostat by Leica
for use with embedding rings.
The inner dimensions of the adapter are 2,3 cm x 2,3 cm.
If you have other questions please feel free contac
Jean,
I work at IMEB, Inc in San Marcos, CA.
I'm very familiar with the instruments listed in the responses to your question.
What it comes down to is cost, closed system or open system (use your reagents
or theirs (Vendors).
The Leica BondMax has it strengths and weaknesses as does the Ventana a
We have the adapter here at IMEB, Inc.
Please go to www.imebinc.com for
our contact info and ask for Brad.
Thank you
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That's why we always keep at least one tall person on staff to reach the top
shelves, etc. :)
Claire
From: histonet-boun...@lists.utsouthwestern.edu on behalf of Judith L. Williams
Sent: Fri 1/30/2009 11:59 AM
To: Charles.Embrey
Cc: histonet
Subject: RE: [Histon
Please tell us what chromogen stains and counterstains you are using, and
the type of mounting medium for the coverslip. This will help us try to
answer your question.
Jan Shivers
UMN VDL
- Original Message -
From: "shazana hilda"
To:
Sent: Monday, February 02, 2009 8:10 PM
Subjec
10. perplexed (Angela Bitting)
histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC and ISH
Hi everyone,
I am currently in the process of evaluating IHC equipment to replace my
old Dako Autostainers. The pathologists I work for are interested in
bringing ISH into our lab. My questions a
A lot depends on the stain chosen. I have used Vector's Nova Red for the last
3 years. I have seen no fading of my older Nova Red slides.
Prof. Allen A. Smith
Barry University School of Podiatric Medicine
Miami Shores, Florida
-Original Message-
From: histonet-boun...@lists.utsouthweste
I have been handling Bouin fixed animal tissue for years. We removed the
Bouins picric acid yellow by submerging our specimens in 70% Alcohol saturated
with Lithium Carbonate. It usually takes overnight. I have had PI's bring me
the samples after rinsing their samples in Lithium Carbonate sol
Hi Angela
There was a problem with the heating pads for a certain batch of Ventana
XT's. We had uneven heating for a week or so until the rep replaced the
entire pad. You can check if your machine is one that was made in the
offending batch. No problems since.
Regards
Piero Nelva
Anatom
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