.
Thanks,
Taylor Rosa
-Original Message-
From: Greg Dobbin
Sent: 07-Mar-2022 11:17 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Fixation time and ISH
Background:
I read on a vendor website that tissues stained using In situ hybridization
should not exceed 32 hours fixation
Background:
I read on a vendor website that tissues stained using In situ hybridization
should not exceed 32 hours fixation. Greater than 32 hours can produce
false negative results because the validated retrieving protocol can be
inadequate.
We in fact have seen evidence of this in our lab
Hi histonetters!
Can anyone tell me what, if any, guidelines there are for fixation time for
animal tissue with potential subsequent IHC stains. I am very familiar
with CAP guidelines for receptor testing with breast cases, but I can't
seem to find much on IHC for animal tissue.
Thanks,
Cristi
Hello Histo-land,
I'm going to start doing FISH in our lab, but I've never done it before. I read
that the PFA fixative solution should be made fresh; however, I have some PFA
that is being stored in the -80 freezer (for less than a year). Does anyone
know if this would be ok to use? And
Hi histonet,
Would like to query about which fixative do you prefer for fresh frozen section
slide before HE staining?
Thanks for your advices
Manahil
HTL/ ASCP
Sent from my iPhone
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Histonet@lists.utsouthwestern.edu
: Tuesday, April 28, 2015 11:11 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Fixation of frozen section
Hi histonet,
Would like to query about which fixative do you prefer for fresh frozen section
slide before HE staining?
Thanks for your advices
Manahil
HTL/ ASCP
Sent from my iPhone
Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Manahil
Sent: Tuesday, April 28, 2015 11:11 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Fixation of frozen section
Hi histonet,
Would like to query
...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Manahil
Sent: Tuesday, April 28, 2015 7:23 PM
To: Keyser Gerald T
Cc: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] Fixation of frozen section
What about the Absolute Methanol?
Sent from my iPhone
On Apr 28
: [Histonet] Fixation of frozen section
You can't use full concentrate alcohol on such specimen, otherwise you will
destroy the cells membrane.
For me I am using 80% Ethanol.
Best Regards,
Jamal Rowaihi Anatomic Pathology Supervisor | Al Borg Medical
Laboratories
-Original Message
What are you placing tissue blocks in before decalcification? We place blocks
for decal in a jar of Bouin's Fixative from any time on Day 1, for specimens
received in 10% NBF, until decalcification starts on Day 2. In other words, we
gross specimens received in 10% NBF, then fix overnight in
We have a brain fixing with phenol formalin, any suggestions on anything
special we need to do with the tissue before processing?
Tanya G. Abbott RT (CSMLS)
Manager Technologist, Histology/Cytology
St. Joseph Medical Center
Reading, PA 19603-0316
ph 610-378-2635
fax 610-898-5871
email:
Has anyone had any experience fixing gelatin scaffolds in glutaraldehyde vapour.
One of the people I am working with is trying to do this and was wondering if
anyone out there has had any experience doing this.
Any suggestions would be greatly appreciated.
Thanks
Mark
Dear Histonetters,
What would the effect be of fixing tissues samples in concentrated
formaldehyde instead of 10% buffered formalin? One of our researchers would
like us to prepare some bones for histology staining which have been fixed
in 37% formaldehyde by mistake and stored for 4 years. I
Using pure formaldehyde as a fixative is not a good idea (you already know
that) and the staining will be affected BUT, on the other hand, IHC will only
require stronger HIER (more time and temperature) but other than that I do
think the epitopes will be detected. So the effects will be mostly
What is the minimum time recommended for fixing routine endoscopy biopsies
before placing them on a short bx run program.
Thank you for your help
Fawn
-
This electronic message may contain information that is
confidential or
hours.
Bye
Gudrun Lang
-Ursprüngliche Nachricht-
Von: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von
ncose...@siumed.edu
Gesendet: Samstag, 19. Oktober 2013 16:51
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet
Hello Histonetters:
Our lab has a resident wanting to start a project involving drug effect on tumor
size. Since he is wanting to compare tumor size at certain time points after
drug treatment (and do IHC staining intensity), he wonders how much affect
varying fixation times will have on the
giuli.zun...@gmail.com
A: histonet@lists.utsouthwestern.edu
Inviato: Mercoledì 13 Marzo 2013 11:31
Oggetto: [Histonet] Fixation
I would like to have some informations about fetal brain fixation..
In general, it is in use the fomalin, but during the cut it is possible to
see that in the deep layer
I would like to have some informations about fetal brain fixation..
In general, it is in use the fomalin, but during the cut it is possible to
see that in the deep layer the formalin doesn't arrive...
So, Could you give some suggestion to improve this technique?!
Thanks in advance
Best
: Wednesday, March 13, 2013 6:31 AM
Subject: [Histonet] Fixation
I would like to have some informations about fetal brain fixation..
In general, it is in use the fomalin, but during the cut it is possible to
see that in the deep layer the formalin doesn't arrive...
So, Could you give some
Rene is correct, formalin fixes slowly so more time is needed. Raising
the concentration of formaldehyde will not help. Adding a small amount
of glutaraldehyde (0.25 to 0.5%) will fix more quickly but may have a
negative effect on immunostaining.
Geoff
On 3/13/2013 6:31 AM, Giulia Zunino
Good Morning-
Scenario: a specimen is received and has been in cytology fixative overnight -
a cell block is made from (very small) tissue picked out.
1.Can this be run on a program without formalin?
2. Does this need to be in formalin before going on the processor - if so
how
@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu
Sent: Wednesday, February 6, 2013 10:42 AM
Subject: [Histonet] fixation
Good Morning-
Scenario: a specimen is received and has been in cytology fixative overnight -
a cell block is made from (very small) tissue picked out.
1. Can this be run
Hello
I need the protocol for fixation, tissue processing and mouse eye sectioning if
you have it can you please send it to me.
thank you
Narjes Baazaoui
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Histonet@lists.utsouthwestern.edu
I agree with Rene.
Geoff
On 12/3/2012 12:00 AM, Daniela Bodemer wrote:
Hi all,
Our tissue processor has been shut down due to a contamination issue and
now all the tissues (mice and rat pelves) collected prior to this
happening have been sitting in 4% PFA. Some tissues more than a week,
Hi all,
Our tissue processor has been shut down due to a contamination issue and
now all the tissues (mice and rat pelves) collected prior to this
happening have been sitting in 4% PFA. Some tissues more than a week,
when we usually fix for 48 hours.
Now we are transferring the tissues to
: Daniela Bodemer daniela.bode...@mcri.edu.au
To: Histonet@lists.utsouthwestern.edu
Sent: Monday, December 3, 2012 12:00 AM
Subject: [Histonet] Fixation time
Hi all,
Our tissue processor has been shut down due to a contamination issue and
now all the tissues (mice and rat pelves) collected prior
Hey fellow histo nuts! Anyone have any good recommendations for fixing breast
tissue? We were wondering about an alcoholic formalin but are unsure if it
will hinder or mess up the SISH results.
We usually end up re-processing the block again just to get a good slide that
is semi readable.
O'Connor ama...@ypii.com
Subject: [Histonet] Fixation for breasts
To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu
Date: Monday, March 19, 2012, 11:36 AM
Hey fellow histo nuts! Anyone have any good recommendations for fixing breast
tissue? We were wondering about
Anyone has a good protocol to fix, process, embed and section mouse eye?
Prefered fixative is formalin. Any suggestions are welcome
Thank you!
Mesru Turkekul
mskcc.org
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Histonet@lists.utsouthwestern.edu
Hello to all,
I have some lactating mouse mammary tissue that was fixed in 10% formalin for
24 hours and then stored in 70% alcohol for 1 day before processing. The
processing is as follows: 80%-30min, 95%-30min, 95%-40 min, 100% X3-30 min.
each, xylene X3-40min. each, and paraffin X3-45min.
I think longer fixation AND processing. Fix for at least 48 hours in NBF
and here is my processing schedule
70% for 45 min
70% for 45 min
95% for 45 min
95% for 45 min
100% for 45 min
100% for 45 min
100% for 45 min
xylene for 45 min
xylene for 45 min
xylene for 45 min
paraffin for 45 min
-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jan
Berry
Sent: 28 January 2011 18:15
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] fixation question for IHC
Is there a difference between using paraformaldehyde
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Margaret Blount
Sent: Monday, January 31, 2011 4:01 AM
To: Jan Berry; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] fixation question for IHC
It is possible to freeze paraformaldehyde in aliquots, so you don't have
to keep
-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jan Berry
Sent: Friday, January 28, 2011 1:15 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] fixation question for IHC
Is there a difference between using paraformaldehyde
Happy Friday to All:
I have started seeing things written that states that the time of
fixation in 10% neutral buffered formalin will be changing to 72 hrs for
breast cases and this will include not only HER2,but also ER/PR
guidelines and these should be releases in 2010. Just wondering if
anyone
Fix in buffered glutaraldehyde for 2 hours, overnight is acceptable.
After multiple rinses, store in buffer in the refrigerator. Change the
buffer once a week, there are things that will grow in cacodylate
buffer. You may complete osmication if you want, then rinse and store in
buffer. I would
Dear all,
I would like to fix and preserve mouse skin tissue for processing for
transmission electron microscopy (TEM) at a later date. I was wondering
whether I can store tissue following fixation overnight in 2.5%
gluteraldehyde in 0.1M sodium cacodylate buffer and, if so, whether it can
be
Hello - A researcher is fixing 7-21 day old chicken embryos in 10% NBF.
After processing they are mushy and almost amorphous. Has anyone any
suggestions for a fixative that will give the embryos more form and still
have the tissue capable of doing IHC? I had thought of 4F1G. Any help is
If the G in 4F1G is glutaraldehyde the glut. will probably kill off the
immunoreactivity (depending on the antigen in question).
How long are the embryos being fixed? Remember, formalin works slowly so
5-7 days may be necessary.-
I would try a formalin + alcohol mix or formalin-alcohol-acetic
Use Bouin's fixative.
René J.
--- On Mon, 8/10/09, Cheryl Crowder ccrow...@vetmed.lsu.edu wrote:
From: Cheryl Crowder ccrow...@vetmed.lsu.edu
Subject: [Histonet] Fixation of embryos
To: Histonet histo...@pathology.swmed.edu
Date: Monday, August 10, 2009, 11:27 AM
Hello - A researcher
Hi histonetters,
Please help. What is difference in procedure for IHC between 10%Formalin fixed
tissue and in Bouin's fixed tissue? I used to use 10%FFPE tissue with Citrate
buffer Antigen Retrieval for my IHC. Do I have to change my protocol for the
Bouin's fixed tissue?
Hope for you soon
Hi Chana,
I'm going to highlight my answers after each of your questions below.
Though I will say that you should contact whomever may be processing the brains
for EM to see what they would prefer. Also ask them which buffer they prefer.
Karnovsky's fix is traditionally made with Sodium
Oops, My highlighted portions didn't make it through the filter. The answers
are after each question.
:-}
Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF)
Core Research Imaging Center
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
Dear Histoneters,
I am doing some IHC on rat cerebellar granular cells (NOT TISSUE) and
I need to fix them with PF.
I have gotten 4 different answers on how to go about this, and I
wanted to run this by the list to see what you think.
1) Fix in COLD, 4% PF in 1x PBS
2) Fix in COLD, 4% PF
Hi, i normally work on tissue.
I think 4% PFA in 0.1M PB is fine. so approach 3.
2009-04-08
TF
发件人: Guillermo Palchik
发送时间: 2009-04-08 05:03:13
收件人: histonet
抄送:
主题: [Histonet] Fixation question - Cerebellar granular cells
Dear Histoneters,
I am doing some IHC on rat cerebellar
Does anyone know if it is possible to fix tissue for IPX in the unbuffered
zinc formalin and then put it on a processor (after it is completely fixed)
with 10% NBF.
Will this undo exactly what you are trying to do? i.e. expose the antigenic
sites
Thanx
Maureen
Hi TF:
Your problem should not be happening.
Sounds like poor perfusion followed by immersing the whole brain (with
the wound deep in the interior) in a small volume of fix at 4 degrees C.
What fixative are you using?
Geoff
TF wrote:
Hi, I just want to fix the damaged brain tissue (2-48
into PFA for 1-2 days.
It should work...
The perfusion is not well in many conditions - in my injury model, part of the
blood supply was interrupted.
2009-01-12
TF
发件人: Geoff McAuliffe
发送时间: 2009-01-12 22:59:22
收件人: tifei
抄送: histonet
主题: Re: [Histonet] Fixation of the damaged brain
Thanks, Dont you think high pressure during perfusion leads to tissue swelling
and disrupted morphology?
2009-01-13
TF
发件人: Charles Scouten
发送时间: 2009-01-13 00:02:40
收件人: ti...@foxmail.com
抄送: histonet
主题: RE: Re: [Histonet] Fixation of the damaged brain tissue
Formaldehyde
Hi, I just want to fix the damaged brain tissue (2-48 hours after I made the
injury).
Because the injury damaged the surrounding blood supply, and after perfusion I
normally got a mass of soft, shapeless tissue surrounding the injury site...
I have also tried post-fixation of 24-48 hours before
Hi,
I was wondering about the effects of using different fixatives in
different orders. For example, some of our embryonic material (chick and
mouse) is fixed initially with 4% PFA and subsequently is exposed to
glutaraldehyde. Chick embryos subject to this are successfully labeled
using IHC
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