[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-----
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Sent: Tuesday, December 14, 2004 10:14 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 13, Issue 20

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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


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------------------------------

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

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------------------------------

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

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------------------------------

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
	
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------------------------------

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

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------------------------------

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

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The information contained in this message may be privileged and/or
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------------------------------

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
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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



------------------------------

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