[Histonet] ihc problem on these Dako HSV-1,HSV-2,HBSAG
Ridhwaan Arries
arrrid702 <@t> skmc.gov.ae
Tue Dec 14 22:26:45 CST 2004
Good morning,
Could any ihc experts out there please help me in solving these problems
1. Dako HSV-1 (clone B0114)
The dilution I used was 1:100-Too much background staining on Dako
HSV CONTROL SLIDES (code #T1150).Slides were HIER and tried without HIER
With the same results.
2. Dako HSV-2 (clone B0116)
The dilution I used was 1:100-Too much background staining on Dako
HSV CONTROL SLIDES (code #T1150).Slides were treated as above.
3. Dako HBsAG (clone B0560)
The dilution used was 1:15,000-No staining at all on Dako HBsAG CONTROL
Slides (code #T1110).Slides were pretreated with and without HIER as well
All enzymes.
Anyone up to advice?
Ridhwaan Arries-Med.Tech./HISTOLOGY(Pentech-South Africa)
-----Original Message-----
From: histonet-request <@t> lists.utsouthwestern.edu [mailto:histonet-request <@t> lists.utsouthwestern.edu]
Sent: Tuesday, December 14, 2004 10:14 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 13, Issue 20
Send Histonet mailing list submissions to
histonet <@t> lists.utsouthwestern.edu
To subscribe or unsubscribe via the World Wide Web, visit
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
or, via email, send a message with subject or body 'help' to
histonet-request <@t> lists.utsouthwestern.edu
You can reach the person managing the list at
histonet-owner <@t> lists.utsouthwestern.edu
When replying, please edit your Subject line so it is more specific
than "Re: Contents of Histonet digest..."
Today's Topics:
1. galectin 4 (Kathy Cormier)
2. FW: [Histonet] Agitation of cassettes (DiCarlo, Margaret)
3. Survey: Histo in Nevada? (RCHIOVETTI <@t> aol.com)
4. CD 68 on murine brain (Favara, Cynthia (NIH/NIAID))
5. RE: CD 68 on murine brain (Elizabeth Chlipala)
6. Histonet: Movat's stain - advice needed! (Nora Berghoff)
7. RE: Microwave Devices and Proposed Guidelines (Bartlett, Jeanine)
8. storage of cryosections (Birthe Schnegelsberg)
9. Collagen IHC in mouse tissue--need antibody recommendations!
(Jack England)
10. Re: storage of cryosections (John Kiernan)
11. RE: IHC ~ Tissue Falling Off Slides (Christine Tambasco)
12. Question to assist a someone off this list (Pamela Marcum)
13. Re: storage of cryosections (Philip Oshel)
14. RE: Microwave Devices and Proposed Guidelines (Bonner, Janet)
15. unscribe (Rita Riddle)
16. Re: storage of cryosections (Gayle Callis)
17. Payscale (Brianna Jackson)
18. Position (Vicki Gauch)
19. Special coatings for slides (Gayle Callis)
20. Varicella-zoster (Cory Collins)
21. MMP-1 in Bouin's fixed tissue (Mark Elliott)
22. Collagen IHC in mouse tissue - need antibody recommendations
(Paula Pierce)
23. RE: Collagen IHC in mouse tissue - need
antibodyrecommendations
(Marshall Terry Dr, Consultant Histopathologist)
----------------------------------------------------------------------
Message: 1
Date: Mon, 13 Dec 2004 13:07:43 -0500
From: Kathy Cormier <cormier <@t> MIT.EDU>
Subject: [Histonet] galectin 4
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <5.2.1.1.2.20041213130230.01557230 <@t> po14.mit.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Hello All,
Hope your day is warm, our temp is rapidly dropping! I have a researcher
who is looking for an antibody to work on FFPE mouse tissue, for galectine
4. Any one using anything that works for FFPE, not just Westerns/flow? :)
Thanks!
Kathy Cormier
Div Comp Medicine
MIT
------------------------------
Message: 2
Date: Mon, 13 Dec 2004 14:01:22 -0500
From: "DiCarlo, Margaret" <MDiCarlo <@t> KaleidaHealth.Org>
Subject: FW: [Histonet] Agitation of cassettes
To: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<DF6AB9298498D31199CF0008C7E6C12010CA66B1 <@t> kalmb02.kaleidahealth.org>
Content-Type: text/plain; charset="iso-8859-1"
Peggy DiCarlo HT (ASCP)
Orthopedics Bone Lab
Buffalo General Hospital
100 High St.
Buffalo, NY 14203
716-859-1293
-----Original Message-----
From: DiCarlo, Margaret
Sent: Friday, November 12, 2004 15:20
To: 'Patsy Ruegg'
Subject: RE: [Histonet] Agitation of cassettes
Patsy,
I too also process most of my bone manually. I agitate on a magnetic stir
plate at room temperature with the bone sitting on a piece of plastic or
something of that nature. I never used it for fixation. I just usually let
it sit in formalin for 5-7 days depending on size and thickness. How much
less time are we talking about, with a bone section that is around 5-8mm in
thickness?
Thanks for your response.
Peggy DiCarlo HT (ASCP)
Orthopedics Bone Lab
Buffalo General Hospital
100 High St.
Buffalo, NY 14203
716-859-1293
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Patsy
Ruegg
Sent: Friday, November 12, 2004 12:36
To: 'Molinari, Betsy'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Agitation of cassettes
I too have always processed most of my tissue by hand (usually bone) and
swear by using a shaker for all steps, fixation, decal, etc. For mass
processing using a tissue processor mine has a stir bar at the bottom of
the tank and I do choose vacuum when ever possible.
Patsy
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Molinari, Betsy
Sent: Friday, November 12, 2004 6:08 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Agitation of cassettes
Hi Fran,
Happy Friday to you! We do put our cassettes on a shaker. I work in a
research lab and have the luxury of time (well, most of the time I do)
to do so. I usually just keep the cassettes on overnight. I always put
my decals on a rotator or shaker it cuts my decal time by quite a bit. I
do not have a publication, but I am someone on this site will! Betsy
Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology
Houston,TX 77030 832-355-6524
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Fran
Lemons
Sent: Thursday, November 11, 2004 2:20 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Agitation of cassettes)
Hello all, it's almost Friday!!
Has anyone out there ever heard of placing the basket of cassettes in
formalin on an agitator to stir the formalin as a way to expedite
fixation? If so, can you direct me to something published on the
subject? Also, has anyone heard of the same thing for bunions in decal
solution? Thanks ahead of time, Fran Walker
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
CONFIDENTIALITY NOTICE:
This email transmission and any documents, files,
or previous e-mail messages attached to it are
confidential and intended solely for the use of the
individual or entity to whom they are addressed.
If you are not the intended recipient, or a person
responsible for delivering it to the intended recipient,
you are hereby notified that any further review,
disclosure, copying, dissemination, distribution, or
use of any of the information contained in or attached
to this e-mail transmission is strictly prohibited.
If you have received this message in error, please
notify the sender immediately by e-mail, discard
any paper copies, and delete all electronic files
of the message. If you are unable to contact the
sender or you are not sure as to whether you
are the intended recipient, please e-mail
ISTSEC <@t> KaleidaHealth.org or call (716) 859-7777.
------------------------------
Message: 3
Date: Mon, 13 Dec 2004 14:38:12 EST
From: RCHIOVETTI <@t> aol.com
Subject: [Histonet] Survey: Histo in Nevada?
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <8c.1c0ae154.2eef49a4 <@t> aol.com>
Content-Type: text/plain; charset="US-ASCII"
Fellow Histonetters,
If you are a practicing histotechnologist in the state of Nevada and if you
can spare about 10-15 minutes to participate in a survey, I would appreciate
hearing from you.
This isn't a salary survey; it's geared more towards the general "health and
welfare" of the profession and the labs in the state (size of labs, number of
docs on staff, case load, instrumentation, usage, service and maintenance
issues and so on).
I need input from *all* histo labs, whether they're hospital labs, derm/Mohs
clinics, veterinary diagnostics labs, reference labs, academic/research labs,
etc.
You can easily complete the survey at your computer, then simply return it to
me as an attachment to an e-mail message. I also have a hard copy version
that I can send to you via snail-mail if you prefer.
Please contact me off-list if you'd like to participate or if you have any
questions.
Thanks in advance. Happy Holidays to everyone!
Cheers,
Robert (Bob) Chiovetti, Ph.D.
Tucson, AZ
------------------------------
Message: 4
Date: Mon, 13 Dec 2004 14:54:17 -0500
From: "Favara, Cynthia (NIH/NIAID)" <cfavara <@t> niaid.nih.gov>
Subject: [Histonet] CD 68 on murine brain
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<F76C9B2DA2FC4C4CA0A18E288BBCBCF70A8DB0C2 <@t> nihexchange24.nih.gov>
Content-Type: text/plain
Has anyone done any CD68 oh FFPE mouse brain?
I will search the archives and do a lit search but have not as of this time,
Thanks,
c
Cynthia Favara
NIAID/NIH/RML/LPVD
903 South 4th Street
Hamilton, MT 59840
406-363-9317
Disclaimer:
The information in this e-mail and any of its attachments is confidential
and may contain sensitive information. It should not be used by anyone who
is not the original intended recipient. If you have received this e-mail in
error please inform the sender and delete it from your mailbox or any other
storage devices. National Institute of Allergy and Infectious Diseases shall
not accept liability for any statements made that are sender's own and not
expressly made on behalf of the NIAID by one of its representatives
------------------------------
Message: 5
Date: Mon, 13 Dec 2004 13:39:04 -0700
From: "Elizabeth Chlipala" <liz <@t> premierlab.com>
Subject: RE: [Histonet] CD 68 on murine brain
To: "'Favara, Cynthia \(NIH/NIAID\)'" <cfavara <@t> niaid.nih.gov>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000201c4e153$c9489990$76d48a80 <@t> AMY>
Content-Type: text/plain; charset="US-ASCII"
Cynthia
I use serotec's antibody F4/80 for macrophages in mouse tissues. I have
not tried it on brain, but I even have done it on formalin fixed, formic
acid decalcifed mouse joints. This antibody will work on FFPE or
paraformaldehye fixed tissue with proteinase K digestion. Here is the
basic protocol.
1:1000 primary 30 minutes room temp
Proteinase K pretreatment - 5 minutes
Rabbit anti-rat 1:400 30 minutes
Envision Rabbit - 30 minutes
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz <@t> premierlab.com
www.premierlab.com
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Favara,
Cynthia (NIH/NIAID)
Sent: Monday, December 13, 2004 12:54 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] CD 68 on murine brain
Has anyone done any CD68 oh FFPE mouse brain?
I will search the archives and do a lit search but have not as of this
time,
Thanks,
c
Cynthia Favara
NIAID/NIH/RML/LPVD
903 South 4th Street
Hamilton, MT 59840
406-363-9317
Disclaimer:
The information in this e-mail and any of its attachments is
confidential
and may contain sensitive information. It should not be used by anyone
who
is not the original intended recipient. If you have received this e-mail
in
error please inform the sender and delete it from your mailbox or any
other
storage devices. National Institute of Allergy and Infectious Diseases
shall
not accept liability for any statements made that are sender's own and
not
expressly made on behalf of the NIAID by one of its representatives
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 6
Date: Mon, 13 Dec 2004 14:52:30 -0600
From: "Nora Berghoff" <NBerghoff <@t> cvm.tamu.edu>
Subject: [Histonet] Histonet: Movat's stain - advice needed!
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <s1bdacc9.048 <@t> cvm.tamu.edu>
Content-Type: text/plain; charset=US-ASCII
Hello,
I have a question about Russell's modification of Movat's pentachrome
stain.
(Ref.: Russell, HK, "A Modification of Movat's Pentachrome stain", Arch
Path, Vol. 94, Aug 1972, 187-191)
I have been staining canine intestinal sections that have been fixed in
10% neutral buffered formalin, embedded in paraffin and cut at 5um.
The stain worked pretty well and gave good results for a while. Last
week I have had the following problem:
After destaining the crocein scarlet-acid fuchsin with 5%
phosphotungstic acid and then rinsing it in 0.5% acetic acid, it kept
destaining the tissue until everything was just very pale red (one slide
was completely destained - no red stain left whatsoever and the other
stains were pale as well).
I always control the destaining under the microscope and stop when
there is still intense red color.
The color kept coming off the slide in the acetic acid solution and
also in the following rinses in absolute alcohol.
I make the solutions (phosphotungstic acid and acetic acid) fresh right
before I need them.
Does anyone have an idea as to what the problem might be? Is the acetic
acid not strong enough/the phosphotungstic acid too strong, etc?? Is it
a pH problem?
I have searched for an answer in histo books, on the internet and in
the histonet archives, but couldn't find anything helpful...
Any advice (even if it is just an idea) would be greatly appreciated!!!
Thank you so much,
Nora Berghoff
Research Assistant
Texas A&M University
College Station, TX
------------------------------
Message: 7
Date: Mon, 13 Dec 2004 17:49:09 -0500
From: "Bartlett, Jeanine" <JQB7 <@t> CDC.GOV>
Subject: RE: [Histonet] Microwave Devices and Proposed Guidelines
To: "Sandy Cheasty" <SCheasty <@t> memorialcare.org>,
<histonet <@t> pathology.swmed.edu>
Message-ID:
<CB857F6460D42E4AAEA195054A25406C050B54EA <@t> m-ncid-2.ncid.cdc.gov>
Content-Type: text/plain; charset="UTF-8"
We have a commercial (for laboratory use) microwave in our main lab. Recently we set up a small, research lab for prion work. I needed a microwave for antigen retrieval but did not have space (or money) for a commercial microwave. Our safety department approved our use of a small, litchen microwave (we got it for less than $100.00 from Target) for this purpose.
Jeanine Bartlett
CDC, Atlanta
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Sandy Cheasty
Sent: Fri 12/10/2004 2:58 PM
To: 'histonet <@t> pathology.swmed.edu'
Cc:
Subject: [Histonet] Microwave Devices and Proposed Guidelines
I know there are thousands of histology labs that use commercial microwaves
safely and have used them safely for years.
Does everyone:
-violate OSHA?
-lie to their lab managers?
There must be some way of continuing to use commercial microwave ovens for
special stains without being subversive. Don't get me wrong, I usually enjoy
being subversive.
Can someone please give me a justification for using a commercial microwave
or tell me where I can get an OSHA approved one for less than $1000?
Thanks again,
Elvis
-----Original Message-----
From: Willis, Donna [mailto:DonnaWillis <@t> texashealth.org]
Sent: Friday, December 10, 2004 7:08 AM
To: Sandy Cheasty; histonet <@t> pathology.swmed.edu
Subject: RE: [Histonet] NCCLS Microwave Device Use Proposed Guideline
It is against OSHA guideline (OSHA 29 CFR 1910.303(b)(2))to use a commercial
microwave (exp. Wal-Mart). The warranty of commercial units state that the
instrument is not to be used for anything except its original use, food
preparation. The OSHA guideline would include any equipment in the lab. If
someone gets injured the lab would be at fault not the manufacture.
Donna Willis
Histology Lab Manager
Harris Methodist Fort Worth,Tx
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sandy
Cheasty
Sent: Friday, December 10, 2004 7:40 AM
To: histonet <@t> pathology.swmed.edu
Subject: [Histonet] NCCLS Microwave Device Use Proposed Guideline
I have the GP28-P guideline and read it but am unsure about its
recommendations or limitations in using a microwave for special stains only.
Is there any consensus among histology labs as to its interpretation? Is it
still OK to use a commercial microwave from Wal-Mart without special exhaust
system as long as you don't microwave formaldehyde, osmium tetroxide, lead
acetate etc. (Paragraph 4.4) and do annual leakage tests?
Thanks!
Elvis
____________________________________________________________________________
__
Confidentiality Notice: This e-mail message, including any attachments, is
for the sole use of the intended recipient(s) and may contain confidential
and privileged information. Any unauthorized review, use, disclosure or
distribution is prohibited. If you are not the intended recipient, please
contact the sender by reply e-mail and destroy all copies of the original
message.
Sign up for your free MemorialCare Medical Information and Access Card at
http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
The information contained in this message and any attachments is intended
only for the use of the individual or entity to which it is addressed, and
may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from
disclosure under applicable law. If you are not the intended recipient, you
are prohibited from copying, distributing, or using the information. Please
contact the sender immediately by return e-mail and delete the original
message from your system.
______________________________________________________________________________
Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message.
Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 8
Date: Mon, 13 Dec 2004 15:23:52 -0600
From: "Birthe Schnegelsberg" <schnegelsberg <@t> xgene.com>
Subject: [Histonet] storage of cryosections
To: <histonet <@t> pathology.swmed.edu>
Message-ID: <NLEHIDDFJOPCHBHFEAONEEBBCAAA.schnegelsberg <@t> xgene.com>
Content-Type: text/plain; charset="iso-8859-1"
HI.
Do I actually have to add Drierite absorbent into the boxes with the cryo
sections in the -20C freezer to avoid freezing artefacts?
Thanks, birthe
------------------------------
Message: 9
Date: Mon, 13 Dec 2004 14:50:10 -1000
From: "Jack England" <joeamateur <@t> hotmail.com>
Subject: [Histonet] Collagen IHC in mouse tissue--need antibody
recommendations!
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <BAY19-F29C373DA946E3819ADB930D4AC0 <@t> phx.gbl>
Content-Type: text/plain; format=flowed
Aloha Histonetters, and happy holidays to all!
We've been trying to do immunostaining for collagen I on some murine-derived
cell lines, with next to no success. I'm almost certain that I've got
collagen in there (PSR shows plenty of red, my poor-man's polarizers make
the red stuff look birefringent, and SEM images show all kinds of fibrous
ECM), but it's not staining for collagen I or III with the antibodies we've
got (both from Sigma, produced in mouse ascites, not tested on mouse
tissue). I suspect that our antibodies don't react with mouse tissue, but
it's possible that the fixation (we use 10% NBF) or the embedding (paraffin,
which does involve heating the tissue) could be denaturing the antigenic
sites. I've tried enzymatic antigen retrieval, and it didn't work...next
step is to get ahold of mouse tails and a cryostat and try for a positive
control.
My question for anyone that can help is, assuming that the problem is that
our antibodies do not react with mouse tissue, can any of you recommend
antibodies to collagen I or III that do react with mouse tissue (preferably
FFPE mouse tissue, since we don't have a cryostat)? I've checked the U of
Iowa hybridoma bank and come up empty, and a Biocompare search gave me one
hit--I'm curious if any of you out there can recommend something beyond that
one.
Any takers?
--With warm regards and aloha,
Jack England
Tissue Genesis, Inc.
http://www.tissuegenesis.com
_________________________________________________________________
On the road to retirement? Check out MSN Life Events for advice on how to
get there! http://lifeevents.msn.com/category.aspx?cid=Retirement
------------------------------
Message: 10
Date: Mon, 13 Dec 2004 20:43:26 -0500
From: John Kiernan <jkiernan <@t> uwo.ca>
Subject: Re: [Histonet] storage of cryosections
To: Birthe Schnegelsberg <schnegelsberg <@t> xgene.com>
Cc: histonet <@t> pathology.swmed.edu
Message-ID: <41BE453E.BA6D31CA <@t> uwo.ca>
Content-Type: text/plain; charset=us-ascii
Freezing artefacts (ice crystal holes) form while
the tissue is being frozen, not during storage of
sections. Once the holes are there, because of
too-slow freezing, nothing can get rid of them.
Drierite (or a similar desiccant) ensures that
the stored sections are in a dry atmosphere.
You should let the box warm to room temperature
before you open it; otherwise water from the air
will condense on the cold sections. If they are of
unfixed tissue this will cause some damage, which may
or may not matter depending on what you intend to
do with the slides.
John Kiernan
London, Canada.
------------------------------------------------
Birthe Schnegelsberg wrote:
>
> HI.
> Do I actually have to add Drierite absorbent into the boxes with the cryo
> sections in the -20C freezer to avoid freezing artefacts?
> Thanks, birthe
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 11
Date: Tue, 14 Dec 2004 01:52:18 +0000
From: "Christine Tambasco" <immrstambo <@t> hotmail.com>
Subject: RE: [Histonet] IHC ~ Tissue Falling Off Slides
To: lpjones <@t> srhs-pa.org, Histonet <@t> lists.utsouthwestern.edu
Message-ID: <BAY12-F1A9870365009B8717B447D2AC0 <@t> phx.gbl>
Content-Type: text/plain; format="flowed"
I use poly-l-lysine slides for immunos (i make them myself) . Also i
use fisher's probe slides and the citra retreival i do in the
microwave. I put the slides in a plastic slide mailer with citra -
microwave on high for 45sec - 1min then I add more citra and microwave
3 minutes at 50% power. It works for me. Good Luck!
Christine Tambasco, HT (ASCP)
St. Mary's Hospital , Amsterdam, New York ph-5188417287
>From: "Jones, Laura" <lpjones <@t> srhs-pa.org>
>To: "Histonet (E-mail)" <Histonet <@t> lists.utsouthwestern.edu>
>Subject: [Histonet] IHC ~ Tissue Falling Off Slides
>Date: Thu, 9 Dec 2004 11:03:38 -0500
>
>Hello fellow Histonetters.
>
>We have suddenly started to have problems with tissue falling off
slides
>during IHC processing. We suspect the problem may be our slides,
although
>we have been using the same ones from Mercedes Medical for a while
and they
>have worked beautifully! We have contacted them, and they are
investigating
>and very kindly replacing our stock, but in the meantime, our
Pathologist
>has asked me to ask all of you where else we can find 25 X 75
silinated
>slides. We use the Zymed ST-5050 automated stainer, and the 1 X 3
slides
>are sometimes a bit to big to fit on the carousel.
>
>Just for additional info, we seem to be experiencing problems mostly
with
>the more alkaline retreival solutions - AR-10 (Biogenex) and EDTA
(Zymed)
>and Trilogy (Cell Marque). We use the Black & Decker steamer, and
have
>adjusted our times in retrieval from 75 minutes (!) to 20 minutes,
per
>Pathologist direction. We have also tried 20 minutes heating, then
adding
>the slides for 20 minutes of boiling, then allowing them to stand for
20-30
>minutes.
>
>Thanks in advance for all of your knowledge!
>
>The Histochicks at Sharon Regional
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 12
Date: Tue, 14 Dec 2004 09:28:24 -0500
From: "Pamela Marcum" <pmarcum <@t> polysciences.com>
Subject: [Histonet] Question to assist a someone off this list
To: <Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000e01c4e1e9$2b4a0f80$7f00a8c0 <@t> PMARCUM2K>
Content-Type: text/plain; charset="iso-8859-1"
Good Morning,
I have had several request for slides coated with different silanes or
charges for people doing cell culture, plastics and some special histology
stains. They are looking for negatively charged slides or special coatings.
Do any of you on HistoNet know anything about these slides like where I can
tell them to go or what type of coatings are available?
I am sorry to ask this however, I don't know where else to go and as
histologist I am more familiar with the traditional coatings and subbing
materials.
Thanks for the help or direction!
Pamela A Marcum
Histology/Microscopy
Product Development Manager
Polysciences, Inc.
400 Valley Road
Warrington, PA 18976
Telephone: 800-523-2575 Ext. 167
215-343-6484 Ext. 167
Fax: 800-343-3291
215-343-0214
------------------------------
Message: 13
Date: Tue, 14 Dec 2004 09:01:13 -0600
From: Philip Oshel <peoshel <@t> wisc.edu>
Subject: Re: [Histonet] storage of cryosections
To: jkiernan <@t> uwo.ca
Cc: Histonet <@t> Pathology.swmed.edu
Message-ID: <p05210602bde4ac2b2b14@[10.25.102.29]>
Content-Type: text/plain; format=flowed; charset=us-ascii
I'm afraid I have to disagree with a portion of John's email: ice
crystal damage can happen in storage.
Ice crystal growth and refreezing occurs at temperatures above about
-60 to -40 deg C, depending on the specifics of the tissue, local
environment, and all the rest of the annoying biological realities we
deal with.
If tissues are frozen rapidly enough to prevent crystal formation, or
to form only tiny crystals, the tissues/cells will experience crystal
growth if they are warmed above the recrystallization temperature.
Much of the damage is actually caused by dehydration as the growing
ice crystals pull free water out of cells and intercellular spaces,
but as crystal growth continues, it can form holes.
The preventative is to store samples in -80 deg C freezers and to
warm them too quickly for recrystallizaton and crystal growth to
occur.
But, there is an even neater trick: store the samples under vacuum in
a -80 freezer with *lots* of desiccant (Lots -- no, more ... pile it
in the chamber. More!*). Something with a high water capacity: best
is 3-4 Angstrom (4 is better than 3, 5 is no good) molecular sieve,
next best is silica gel. Leave the samples like this for a few days
(hours to weeks, depending on sample size and number -- it's
empirical, try it and see), and they will be nicely freeze-dried by
vacuum sublimation. They can then be stored at any temperature with
desiccant with no worries about ice crystals, or much of anything.
On exposure to air, the samples will immediately start rehydrating,
so they *must* be at room temperature before removing them from the
desiccant chamber, and processed right away.
Caveat: I don't think this would adversely affect antigen epitopes,
but this should be tested.
Phil
* If you think you have enough, you don't.
>Freezing artefacts (ice crystal holes) form while
>the tissue is being frozen, not during storage of
>sections. Once the holes are there, because of
>too-slow freezing, nothing can get rid of them.
>
>Drierite (or a similar desiccant) ensures that
>the stored sections are in a dry atmosphere.
>You should let the box warm to room temperature
>before you open it; otherwise water from the air
>will condense on the cold sections. If they are of
>unfixed tissue this will cause some damage, which may
>or may not matter depending on what you intend to
>do with the slides.
>
>John Kiernan
>London, Canada.
>------------------------------------------------
>Birthe Schnegelsberg wrote:
>>
>> HI.
>> Do I actually have to add Drierite absorbent into the boxes with the cryo
>> sections in the -20C freezer to avoid freezing artefacts?
>> Thanks, birthe
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
--
Philip Oshel
Supervisor, BBPIC microscopy facility
Department of Animal Sciences
University of Wisconsin
1675 Observatory Drive
Madison, WI 53706
voice: (608) 263-4162
fax: (608) 262-5157 (dept. fax)
------------------------------
Message: 14
Date: Tue, 14 Dec 2004 10:44:22 -0500
From: "Bonner, Janet" <Janet.Bonner <@t> FLHOSP.ORG>
Subject: RE: [Histonet] Microwave Devices and Proposed Guidelines
To: "'Sandy Cheasty'" <SCheasty <@t> memorialcare.org>,
"'histonet <@t> pathology.swmed.edu'" <histonet <@t> pathology.swmed.edu>
Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4127 <@t> fh2k093.fhmis.net>
Content-Type: text/plain; charset="iso-8859-1"
I've found that the issue of commercial vs industrial microwaves boils down
to a ventilation problem( possible explosion or respiratory problem on a
"stain" becoming an aerosol) as well as QA as far as temperature and time
regulation. The microwave should be vented (ours is in the back) to the
outside or a fume hood.
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Sandy
Cheasty
Sent: Monday, December 13, 2004 11:48 AM
To: 'histonet <@t> pathology.swmed.edu'
Subject: [Histonet] Microwave Devices and Proposed Guidelines
If you receive any info on this would you please let me know. We
are currently using a microwave from Wal-Mart. There has got to be some
kind of exception somewhere. What about labs that could not justify a lab
microwave but has a commercial one that works just as good for special
stains only. Thanks, amy
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Sandy
Cheasty
Sent: Friday, December 10, 2004 2:58 PM
To: 'histonet <@t> pathology.swmed.edu'
Subject: [Histonet] Microwave Devices and Proposed Guidelines
I know there are thousands of histology labs that use commercial microwaves
safely and have used them safely for years.
Does everyone:
-violate OSHA?
-lie to their lab managers?
There must be some way of continuing to use commercial microwave ovens for
special stains without being subversive. Don't get me wrong, I usually enjoy
being subversive.
Can someone please give me a justification for using a commercial microwave
or tell me where I can get an OSHA approved one for less than $1000?
Thanks again,
Elvis
-----Original Message-----
From: Willis, Donna [mailto:DonnaWillis <@t> texashealth.org]
Sent: Friday, December 10, 2004 7:08 AM
To: Sandy Cheasty; histonet <@t> pathology.swmed.edu
Subject: RE: [Histonet] NCCLS Microwave Device Use Proposed Guideline
It is against OSHA guideline (OSHA 29 CFR 1910.303(b)(2))to use a commercial
microwave (exp. Wal-Mart). The warranty of commercial units state that the
instrument is not to be used for anything except its original use, food
preparation. The OSHA guideline would include any equipment in the lab. If
someone gets injured the lab would be at fault not the manufacture.
Donna Willis
Histology Lab Manager
Harris Methodist Fort Worth,Tx
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sandy
Cheasty
Sent: Friday, December 10, 2004 7:40 AM
To: histonet <@t> pathology.swmed.edu
Subject: [Histonet] NCCLS Microwave Device Use Proposed Guideline
I have the GP28-P guideline and read it but am unsure about its
recommendations or limitations in using a microwave for special stains only.
Is there any consensus among histology labs as to its interpretation? Is it
still OK to use a commercial microwave from Wal-Mart without special exhaust
system as long as you don't microwave formaldehyde, osmium tetroxide, lead
acetate etc. (Paragraph 4.4) and do annual leakage tests?
Thanks!
Elvis
____________________________________________________________________________
__
Confidentiality Notice: This e-mail message, including any attachments, is
for the sole use of the intended recipient(s) and may contain confidential
and privileged information. Any unauthorized review, use, disclosure or
distribution is prohibited. If you are not the intended recipient, please
contact the sender by reply e-mail and destroy all copies of the original
message.
Sign up for your free MemorialCare Medical Information and Access Card at
http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
The information contained in this message and any attachments is intended
only for the use of the individual or entity to which it is addressed, and
may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from
disclosure under applicable law. If you are not the intended recipient, you
are prohibited from copying, distributing, or using the information. Please
contact the sender immediately by return e-mail and delete the original
message from your system.
____________________________________________________________________________
__
Confidentiality Notice: This e-mail message, including any attachments, is
for the sole use of the intended recipient(s) and may contain confidential
and privileged information. Any unauthorized review, use, disclosure or
distribution is prohibited. If you are not the intended recipient, please
contact the sender by reply e-mail and destroy all copies of the original
message.
Sign up for your free MemorialCare Medical Information and Access Card at
http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Note: The information contained in this message may be privileged and
confidential and protected from disclosure. If the reader of this
message is not the intended recipient, or an employee or an agent
responsible for delivering this message to the intended recipient, you
are hereby notified that any dissemination, distribution or copying
of this communication is strictly prohibited. If you have received
this communication in error, please notify us immediately by replying
to the message and deleting it from your computer. Thank you.
____________________________________________________________________________
__
Confidentiality Notice: This e-mail message, including any attachments, is
for the sole use of the intended recipient(s) and may contain confidential
and privileged information. Any unauthorized review, use, disclosure or
distribution is prohibited. If you are not the intended recipient, please
contact the sender by reply e-mail and destroy all copies of the original
message.
Sign up for your free MemorialCare Medical Information and Access Card at
http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
The information contained in this message may be privileged and/or
confidential and protected from disclosure. If the reader of this message
is not the intended recipient or agent responsible for delivering this
message to the intended recipient, you are hereby notified that any
dissemination, distribution or copying of this communication is strictly
prohibited. If you have received this communication in error, please notify
the sender immediately by replying to this message and deleting the material
from any computer.
------------------------------
Message: 15
Date: Tue, 14 Dec 2004 10:56:25 -0500
From: Rita Riddle <RitaR <@t> lexhealth.org>
Subject: [Histonet] unscribe
To: "'Histonet <@t> lists.utsouthwestern.edu'"
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<CA70329E0BA39C4A974EDC102ADCE79E058021F8 <@t> mail.lmc.lexhealth.org>
Content-Type: text/plain
_________________________________
This email and any files transmitted with it may contain PRIVILEGED or
CONFIDENTIAL information and may be read or used only by the intended
recipient. If you are not the intended recipient of the email or any of its
attachments, please be advised that you have received this email in error
and that any use, dissemination, distribution, forwarding, printing, or
copying of this email or any attached files is strictly prohibited. If you
have received this email in error, please immediately purge it and all
attachments and notify the sender by reply email or contact the sender at
the number listed above if one is provided.
------------------------------
Message: 16
Date: Tue, 14 Dec 2004 09:04:37 -0700
From: Gayle Callis <gcallis <@t> montana.edu>
Subject: Re: [Histonet] storage of cryosections
To: "Birthe Schnegelsberg" <schnegelsberg <@t> xgene.com>,
Histonet <@t> lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20041214084115.01b0b540 <@t> gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Birthe,
Initially, how you snap freeze is the first defense against freezing
artefacts. Frozen section stored with a dessicant is a good idea keep the
sections dry during storage. The storage container must stay closed to
equilibrate FS to room temperature. Water condensation is the enemy. Be
sure to air dry the sections before going into storage also. A black
plastic 25 slide capacity box works well. Also, opening a box directly
from freezer and taking a few sections out, then returning unused sections
to freezer must be avoided due FS freeze thaw. Put as many slides as you
need per storage container for a staining session.
Drierite is anhydrous calcium chloride and has a tendency to exfoliate fine
CaCl2dust all over everything including your sections which bothered
me. Try 10 to 18 mesh silica gel, (Fisher# S161-500). The little beads
have blue indicator so you know when dessicant is depleted. Put Silica gel
in a nylon processing bags (Thermo Electron or StatLab) or embedding bags
aka tea bags from Fisher, fold over top, and staple on fold the keep bags
closed.
Temperature is critical for storage - if you have -80C available, use that
instead of -20C, and NEVER store sections in a self defrosting freezer
found in a refrigerator, freeze dry cycle is very damaging. Hopefully you
can store FS at temperature lower than
-20C to preserve antigenicity over a long period of time. It may work for
short term storage and robust antigens.
At 02:23 PM 12/13/2004, you wrote:
>HI.
>Do I actually have to add Drierite absorbent into the boxes with the cryo
>sections in the -20C freezer to avoid freezing artefacts?
>Thanks, birthe
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
------------------------------
Message: 17
Date: Tue, 14 Dec 2004 09:08:09 -0700
From: "Brianna Jackson" <bjackson <@t> unipathllc.com>
Subject: [Histonet] Payscale
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <auto-000019425858 <@t> fe.mail.megapathdsl.net>
Content-Type: text/plain; charset="us-ascii"
Hello,
Can histotechs working in labs with separate IHC and Histo departments tell
me if you have the same payscale? I've been told that IHC techs (even HTL
certified) cannot be paid the same as production bench techs. I was just
wondering if this is how it works in other labs.
Thanks for your time,
Brianna Jackson, BS, QIHC
Denver, CO
303-512-2220
------------------------------
Message: 18
Date: Tue, 14 Dec 2004 11:09:48 -0500
From: "Vicki Gauch" <GauchV <@t> mail.amc.edu>
Subject: [Histonet] Position
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <s1beca1e.082 <@t> amc03.amc.edu>
Content-Type: text/plain; charset=US-ASCII
I have a tech with 10 years of experience who will be relocating to the
Scottsdale,Arizona region who is seeking a Histotech position in that
area. Does anyone know of any openings? Please send any correspondence
to this e-mail address as she is not currently able to access the
Histonet.
Thank you for your help.
Vicki Gauch
Albany Medical Center
Albany, NY
------------------------------
Message: 19
Date: Tue, 14 Dec 2004 09:20:04 -0700
From: Gayle Callis <gcallis <@t> montana.edu>
Subject: [Histonet] Special coatings for slides
To: "Pamela Marcum" <pmarcum <@t> polysciences.com>,
Histonet <@t> lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20041214091203.01b4e368 <@t> gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Pam,
I believe Erie Scientific is doing some of this from what my Erie rep tells
me. Erie website has 800 number for tech services. They have some new
things for Microarray work. Also try Grace Bio-Labs, they have a website,
and you can call them for detailed discussion.
Be prepared for some pricey items.
At 07:28 AM 12/14/2004, you wrote:
>Good Morning,
>
>I have had several request for slides coated with different silanes or
>charges for people doing cell culture, plastics and some special histology
>stains. They are looking for negatively charged slides or special coatings.
>Do any of you on HistoNet know anything about these slides like where I can
>tell them to go or what type of coatings are available?
>
>I am sorry to ask this however, I don't know where else to go and as
>histologist I am more familiar with the traditional coatings and subbing
>materials.
>
>Thanks for the help or direction!
>
>Pamela A Marcum
>Histology/Microscopy
>Product Development Manager
>Polysciences, Inc.
>400 Valley Road
>Warrington, PA 18976
>Telephone: 800-523-2575 Ext. 167
> 215-343-6484 Ext. 167
>
>Fax: 800-343-3291
> 215-343-0214
>
>
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
------------------------------
Message: 20
Date: Tue, 14 Dec 2004 10:21:33 -0600
From: "Cory Collins" <CCollins <@t> propathlab.com>
Subject: [Histonet] Varicella-zoster
To: <histonet <@t> lists.utsouthwestern.edu.>
Message-ID:
<C2E3B94F4E416847A4A9C904ABC3498903A4E5 <@t> mail.propathlab.com>
Content-Type: text/plain; charset="iso-8859-1"
Howdy!! I work for a reference lab in Dallas and we are having trouble
finding control tissue for Varicella-zoster. Does anyone know of a vendor
that sells control slides for Varcilla-zoster? Or maybe know someone that
has some extra paraffin embedded tissue that is positive for
Varicella-zoster.
Thanks,
Cory
Cory Collins, HT (ASCP) QIHC
Immunohistochemistry Supervisor
ProPath
8267 Elmbrook Drive Suite 100
Dallas, TX 75247
214-638-2000 ext 2027
214-237-1730 Fax
To learn more about ProPath, please visit http://www.ProPathLab.com
______________________________________________________________________________
This e-mail may contain confidential or privileged information. If you think you have received this e-mail
in error, please advise the sender by reply e-mail and then delete this e-mail immediately.
------------------------------
Message: 21
Date: Tue, 14 Dec 2004 09:19:47 -0800
From: "Mark Elliott" <MElliott <@t> mrl.ubc.ca>
Subject: [Histonet] MMP-1 in Bouin's fixed tissue
To: <Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <s1beb043.094 <@t> mail.mrl.ubc.ca>
Content-Type: text/plain; charset=US-ASCII
Has anyone had any experience staining for MMP-1 in Bouin's fixed human
tissues? One of the Hospital's we collaborate with fixes their lungs in
Bouin's. We have tried staining with no antigen retrieval and
with/without overnight incubation in primary. Have also tried antigen
retrieval using citra pH 6 in 95 C water bath and in autoclave with no
luck. We are using MAB 1346 from Chemicon followed by APAAP technique.
Are going to try high pH antigen retrieval and possibly trypsin but
would appreciate any other suggestions.
Thanks
Mark Elliott
iCAPTURE Centre
Vancouver BC
------------------------------
Message: 22
Date: Tue, 14 Dec 2004 09:34:31 -0800 (PST)
From: Paula Pierce <contact <@t> excaliburpathology.com>
Subject: [Histonet] Collagen IHC in mouse tissue - need antibody
recommendations
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <20041214173431.64936.qmail <@t> web50308.mail.yahoo.com>
Content-Type: text/plain; charset=us-ascii
Have you tried just a histochemical stain for collagen such as Masson's Trichrome?
Paula Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
631 N. Broadway
Moore, OK 73160
405-759-3953
contact <@t> excaliburpathology.com
www.excaliburpathology.com
------------------------------
Message: 23
Date: Tue, 14 Dec 2004 17:38:18 -0000
From: "Marshall Terry Dr, Consultant Histopathologist"
<Terry.Marshall <@t> rothgen.nhs.uk>
Subject: RE: [Histonet] Collagen IHC in mouse tissue - need
antibodyrecommendations
To: "Paula Pierce" <contact <@t> excaliburpathology.com>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<FE2DB935F8BBB546B8A1BBF3459C5A1F02DC9FA4 <@t> LIL.xRothGen.nhs.uk>
Content-Type: text/plain; charset="iso-8859-1"
Masson - histochemical?
Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path
Consultant Pathologist
Rotherham General Hospital
South Yorkshire
England
terry.marshall <@t> rothgen.nhs.uk
-----Original Message-----
From: Paula Pierce [mailto:contact <@t> excaliburpathology.com]
Sent: 14 December 2004 17:35
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Collagen IHC in mouse tissue - need
antibodyrecommendations
Have you tried just a histochemical stain for collagen such as Masson's Trichrome?
Paula Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
631 N. Broadway
Moore, OK 73160
405-759-3953
contact <@t> excaliburpathology.com
www.excaliburpathology.com
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
End of Histonet Digest, Vol 13, Issue 20
****************************************
More information about the Histonet
mailing list