Hello Amos,
I have used the Biocare Medical Goat HRP polymer Kit, which detects Goat
primary antibodies for use on Mouse, Rat, and Human tissue. Catalog# is
GHP516G. Be sure to use some sort of detergent in your wash buffer to
prevent background from ionic interactions on the slides. I use 0.05%
Tween 20 in my wash buffer solution for any Biocare HRP polymer product
and the last step prior to the substrate chromagen should always be a
rinse in deionized water. Coincidently, my wash buffer is also my
antibody diluent. Hope that this helps. Surely, you could by there
special TBS buffer but it's more economical for me to make my own and
add the Tween 20.
Carmen Wynn, M.S., Senior Scientist
Astellas Research Institute of America, LLC. (ARIA)
Illinois Science and Technology Park
8045 Lamon Ave
Skokie, IL 60077
Direct: 847-933-7419
Main: 847-933-7400
Fax: 847-933-7401
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
histonet-request@lists.utsouthwestern.edu
Sent: Wednesday, May 07, 2008 7:39 PM
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 54, Issue 9
Send Histonet mailing list submissions to
histonet@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
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You can reach the person managing the list at
histonet-owner@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. Too many Techs... (Bonner, Janet)
2. RE: B-plus question (Weber, Susan (VHACLE))
3. Re: cryoprotection (John Kiernan)
4. Lot numbers (Webb, Dorothy L)
5. c-kit (CD117) rat and mouse tissue
(michelle.schwab-macdonald@novartis.com)
6. Shirley Phua is out-of-office ... (Shirley PHUA)
7. RE: GLP question (Trajkovic, Dusko)
8. RE: Lot numbers. . (Henry, Charlene)
9. RE: B-plus question. . (Henry, Charlene)
10. Immunohistotechnologist position open at Kansas State Univ.
(Shelly Christenson)
11. RE: Lot numbers. . (Rene J Buesa)
12. RE: Lot numbers (Bonner, Janet)
13. RE: c-kit (CD117) rat and mouse tissue (Wright, Clarissa B CIV)
14. histonet@lists.utsouthwestern.edu (zodiac29@comcast.net)
15. glassware cleaners (Laurie Colbert)
16. RE: More on [Histonet] Looking for detailed protocol to stain
for PASin liver sections (paraffin) (Tony Henwood)
17. RE: More on [Histonet] Looking for detailed protocol to stain
for PASin liver sections (paraffin) (Tony Henwood)
18. Re: Re; Decal Process From the Past (Akemi Allison-Tacha)
19. RE: Too many Techs... (jstaruk)
20. Polymer IHC Detection (Amos Brooks)
----------------------------------------------------------------------
Message: 1
Date: Wed, 7 May 2008 13:06:17 -0400
From: "Bonner, Janet"
Subject: [Histonet] Too many Techs...
To: "Anne van Binsbergen" , "barbara carter"
,
histonet-request@lists.utsouthwestern.edu,
"Histonet"
Message-ID:
<5F31F38C96781A4FBE3196EBC22D47807F2621@fhosxchmb006.ADVENTISTCORP.NET>
Content-Type: text/plain; charset=iso-8859-1
10,000cases? 3300 cases per tech per year, 100 cases per tech per
day? At least 300 blocks min. per day per tech.
CAP has guidelines for justifying positions. I would also have the
techs (like they need something else to do) keep track of their time for
a month. It sounds as if three techs would minimally handle your lab.
Not to mention two months worth of vacation/sick time taken per year.
Also - what is the boss seeing? Coffee breaks? Long lunch breaks?
We have 22 Techs, three shifts, six days per week, 50,000 Surgical
cases. It takes two techs worth just to cover time off per year. We do
not attend procedures and we buy our reagents.
Good luck.
________________________________
From: histonet-bounces@lists.utsouthwestern.edu on behalf of Anne van
Binsbergen
Sent: Wed 5/7/2008 12:09 PM
To: barbara carter; histonet-request@lists.utsouthwestern.edu; Histonet
Subject: Re: [Histonet] Histology Coordinator Position in Kenosha
Wisconsin
i have a few questions for you if i may.
you have 3 techs serving 3 paths and 10 000 a year
apart from the basic routine histo operations, do your 3 techs
- do any grossing
- attend at renal biopsy collection
- do frozen sections
- take macro photos
- do slide and block filing
- source cases for sendout (consults/reviews) or do any archive
searches
- wash glassware,
- do specimen discards,
- make up formalin
i am facing pressure from my (non-histo) boss who is comparing my lab
and
staff to 'other modern facilities' and he now tells me i need to cut my
staff in half!!
i need ammo and numbers from others to build my defence
cheers
Annie
2008/5/7 barbara carter :
>
> I am leaving my position as Coordinator at United Hospital System and
am
> moving out of state and trying to help them recruit a new coordinator
to
> take my place.
>
> There is a position open at United Hospital System at the Kenosha
Campus
> in Kenosha Wisconsin for a Histology Coordinator. The hospital
receives
> about 10,000 surgical specimen a year with 3 Pathologists and 3
Histologist.
>
> Anyone interested please contact Sue Wergin Laboratory Supervisor at
> 262-656-5603.
>
>
>
>
> _________________________________________________________________
> Windows Live SkyDrive lets you share files with faraway friends.
>
>
http://www.windowslive.com/skydrive/overview.html?ocid=TXT_TAGLM_WL_Refr
esh_skydrive_052008_______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
--
Anne (van Binsbergen) Hope
Abu Dhabi
UAE
_______________________________________________
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------------------------------
Message: 2
Date: Wed, 7 May 2008 13:09:45 -0400
From: "Weber, Susan (VHACLE)"
Subject: RE: [Histonet] B-plus question
To: "Paul Bradbury" , "karen adams"
, "HistoNet Server"
Message-ID:
<16C83872A53F4346AA9C3A18E3A3AAB903F76D96@VHAV10MSGA1.v10.med.va.gov>
Content-Type: text/plain; charset="us-ascii"
I may have missed this, but may I have the name of the Vendor you
purchase your B-plus from. Thanks.
Susan M Weber HT(ASCP)
Histology Supervisor
Louis Stokes Cleveland VA Medical Center
10701 East Blvd
Cleveland, Ohio 44106
(216) 791-3800 X6154
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paul
Bradbury
Sent: Tuesday, May 06, 2008 9:53 AM
To: karen adams; HistoNet Server
Subject: Re: [Histonet] B-plus question
Hi Karen,
I have been using the same sequence of reagents for several years with
great success. We routinely fix bone marrow cores for 3 hours in B-plus,
rinse in water, decalcify, rinse again, and put the cassette in with all
the other tissues for processing. B-plus contains formaldehyde anyway,
so your are not introducing a different reagent when you transfer them
to formalin during processing.
B-plus gives much better cytological detail than formalin. Nuclear
detail is crisper, granules are better preserved, hemosiderin is not
effected. CD-3, CD-20, kappa and lambda, etc. all work beautifully after
B-plus. It also has the advantage of not producing an fixation artefact
pigment like B-5 does. I think you will be happy with your decision to
change.
Paul Bradbury
Kamloops, Canada
karen adams wrote:
> Hello all....we are changing bone marrow fixative from formalin to
B-plus.
> If we fix for the required time in B-plu and then process on the VIP
w/ the
> other specimens using formalin are there any effects on the tissue
going
> from B-plus to formalin??
> Thank you in advance
>
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 3
Date: Wed, 07 May 2008 13:24:29 -0400
From: John Kiernan
Subject: Re: [Histonet] cryoprotection
To: Teisha Robertson
Cc: histonet@lists.utsouthwestern.edu
Message-ID:
Content-Type: text/plain; charset="us-ascii"
Transfer the brain, after adequate fixation in buffered
forma brain sin cutting sections. should still freeze as
q on a metal cryostat chuck, compound for adhesion, then
stand the chu CO2 and either acetone or alcohol. Mount the c in
the cryostat and wait for its temperature
equilibrate. This isn't the fastest way to freeze thing but
it's good enough for cryoprotected formaldehyde-fixed rodent
brain UWO
----- Or <tshrobertso May 5, 2008
12:41< cryoprotection
To: histonet@li
sts.utsouthwestern.edu
> i am trying to
cryopr
> brain in sucrose. can you please
to
prevent holes in my sample?
>
---------------------------------
> Be
a
> ______ _______________________ 5F
_________________
>
Hist Histonet@lists.utsouthwestern.ed
http://lists.utsouthwestern.edu/mailman/listinfo
------------------------------
Message: 4
Date: Wed, 07 May 2008 12:56:24 -0500
From: "Webb, Dorothy L"
Subject: [Histonet] Lot numbers
To: histonet@lists.utsouthwestern.edu
Message-ID:
<0E394B648E5284478A6CCB78E5AFDA27056356BB@hpes1.HealthPartners.int>
Content-Type: text/plain; charset="us-ascii"
Does everyone keep track of each reagent lot number, including, water,
formalin, alcohols, etc. as it is opened and used? What about recycled
products? I am trying to come up with a logsheet and would appreciate
any help in this area!
Also, we have recently run into problems with our freezing compound
coming off of the chucks during frozen sectioning. Any suggestions
and/or what is the method of freezing tissue that everyone uses for
their cryostat sections? Thanks fellow histotechs for helping me with
these matters that I am coming up empty on!!!!!!!!!!!!!!!!!!!!!
Dorothy Webb, HT (ASCP)
Histology Technical Supervisor
Regions Hospital, Pathology Department
640 Jackson Street, Saint Paul, MN 55101-2595
Phone: 651-254-2962
Fax: 651-254-2741
Regions Hospital is part of the HealthPartners family of care
________________________________________
This e-mail and any files transmitted with it are confidential and are
intended solely for the use of the individual or entity to whom they are
addressed. If you are not the intended recipient or the individual
responsible for delivering the e-mail to the intended recipient, please
be advised that you have received this e-mail in error and that any use,
dissemination, forwarding, printing, or copying of this e-mail is
strictly prohibited.
If you have received this e-mail in error, please immediately notify the
HealthPartners Support Center by telephone at (952) 967-6600. You will
be reimbursed for reasonable costs incurred in notifying us.
------------------------------
Message: 5
Date: Wed, 7 May 2008 13:56:46 -0400
From: michelle.schwab-macdonald@novartis.com
Subject: [Histonet] c-kit (CD117) rat and mouse tissue
To: histonet@lists.utsouthwestern.edu,
histonet-bounces@lists.utsouthwestern.edu
Message-ID:
Content-Type: text/plain; charset="US-ASCII"
I am looking for a c-kit Ab that will stain FFPE rat or mouse tissue. I
have heard that Dako has one for rat. Is anyone using it and willing to
share protocol information or does anyone have an Ab suggestion?
Thanks in Advance!! :-)
Michelle Schwab-MacDonald HT(ASCP)
Novartis, Boston, MA
------------------------------
Message: 6
Date: Thu, 8 May 2008 02:02:51 +0800
From: Shirley PHUA
Subject: [Histonet] Shirley Phua is out-of-office ...
To: histonet
Message-ID:
Content-Type: text/plain; charset=US-ASCII
I will be out of the office from 08-05-2008 to 09-05-2008.
I'll be away on 08 May 2008.
Pathologists:
I will process your requests when I return. If urgent, please forward
your
email to Henry_Kyaw@hsa.gov.sg
------------------------------
Message: 7
Date: Wed, 7 May 2008 11:16:21 -0700
From: "Trajkovic, Dusko"
Subject: RE: [Histonet] GLP question
To: "Jackie M O'Connor"
Cc: histonet@lists.utsouthwestern.edu,
histonet-bounces@lists.utsouthwestern.edu, Linke_Noelle
Message-ID:
<3AD0BD3142459B4E9B12CBEAFF2B89B2070C2917@lajamrexm01.amer.pfizer.com>
Content-Type: text/plain; charset="us-ascii"
If our lab had the same operation as Jackie O., then that is the way I
would also perform the GLP tracking. However, our set up is similar to
Kim Merriam, where each Histotech is responsible for their own study or
work assigned. One person will do the trimming, embedding, sectioning,
staining and self QC. I don't consider my slides as final product until
I sign off on the QC portion, have someone else verify and initial that
my numbers are correct (blocks and slides match and are all accounted
for) then hand the slides to the pathologist.
During my QC process, if I find that there are imperfections in the
sections (this is hypothetical, since things like that never happen to
me) such as folds, knife marks, floaters, etc. I go back to recut the
slide, restain and QC again. If I am satisfied with my new freshly
coverslipped slide, I sign off the animal and submit my slides. The
slide that I discarded is nothing more than any other section that I
discarded during my microtome sectioning before deciding that I am deep
enough in the block and it's time to collect my ribbon and place on the
water bath.
This is my story, and my opinion on the GLP process. I'm sure that
someone else might have some issues with my process, but every situation
is different. Having worked in a GLP lab for quite a few years, never
had any issues with FDA inspectors regarding any work that I have
submitted.
Dusko Trajkovic
Pfizer Inc, La Jolla
________________________________
From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com]
Sent: Wednesday, May 07, 2008 5:39 AM
To: Trajkovic, Dusko
Cc: histonet@lists.utsouthwestern.edu;
histonet-bounces@lists.utsouthwestern.edu; Kim Merriam; Linke_Noelle
Subject: RE: [Histonet] GLP question
My lab has the capability of producing 600+ slides per day. When we cut
initial slides, the microtomy is documented by the microtomist at the
time. Staining is subsequently documented - since it may be performed
by another technician. Once documentation of a procedural step is
made, that slide is already raw datum. If a recut is needed at QC, the
reason for the recut is also documented, and the original slide is
retained. The recut is subsequently documented, as well as whether or
not the desired goal (missing mammary) was achieved. Gotta love the
world of GLP. We may be documenting this to death - but it works
quite well.
------------------------------
Message: 8
Date: Wed, 7 May 2008 14:53:42 -0500
From: "Henry, Charlene"
Subject: RE: [Histonet] Lot numbers. .
To: "'Webb, Dorothy L'" ,
"histonet@lists.utsouthwestern.edu"
Message-ID:
<03E1F5968F60C5448635D49D38B283ED797B597D@SJMEMXMBS11.stjude.sjcrh.local
>
Content-Type: text/plain; charset="us-ascii"
We had put the tracking of lot numbers and expiration dates of reagents
(alcohol, xylene, methanol, formalin etc) into place just before our
last CAP inspection and I'm glad we did because we would have been sited
with a deficiency. I don't think that it is on the AP Checklist but it
is on the General Checklist. The inspector asking me about the reagent
lot numbers and expiration dates was a chemistry person and I tried to
explain to her that the volume of reagents that a Histology Lab goes
through in comparison with a Chemistry Lab is quite different. I told
her that a Histology Lab would go through more reagents than the rest of
all the clinical labs put together.
I would think it would be impossible to track the lot number of recycled
alcohols.
We don't track the lot numbers of water because we have a Millipore
system. We do track the lot number of formalin as it comes in and is put
into use; however we do not track which lot number of formalin is used
on individual cases.
My experience with the compound coming off chucks during frozen sections
has been when the chuck is too cold so we do not store our chucks in the
cryostat but keep them at room temperature. It really does not add that
much time when freezing tissue.
Charlene Henry HT (ASCP), QIHC
Anatomic Pathology Section Head
Department of Pathology
St. Jude Children's Research Hospital
901-495-3191
fax 901-495-3100
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Webb,
Dorothy L
Sent: Wednesday, May 07, 2008 12:56 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Lot numbers. .
Does everyone keep track of each reagent lot number, including, water,
formalin, alcohols, etc. as it is opened and used? What about recycled
products? I am trying to come up with a logsheet and would appreciate
any help in this area!
Also, we have recently run into problems with our freezing compound
coming off of the chucks during frozen sectioning. Any suggestions
and/or what is the method of freezing tissue that everyone uses for
their cryostat sections? Thanks fellow histotechs for helping me with
these matters that I am coming up empty on!!!!!!!!!!!!!!!!!!!!!
Dorothy Webb, HT (ASCP)
Histology Technical Supervisor
Regions Hospital, Pathology Department
640 Jackson Street, Saint Paul, MN 55101-2595
Phone: 651-254-2962
Fax: 651-254-2741
Regions Hospital is part of the HealthPartners family of care
________________________________________
This e-mail and any files transmitted with it are confidential and are
intended solely for the use of the individual or entity to whom they are
addressed. If you are not the intended recipient or the individual
responsible for delivering the e-mail to the intended recipient, please
be advised that you have received this e-mail in error and that any use,
dissemination, forwarding, printing, or copying of this e-mail is
strictly prohibited.
If you have received this e-mail in error, please immediately notify the
HealthPartners Support Center by telephone at (952) 967-6600. You will
be reimbursed for reasonable costs incurred in notifying us.
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 9
Date: Wed, 7 May 2008 14:58:49 -0500
From: "Henry, Charlene"
Subject: RE: [Histonet] B-plus question. .
To: "'Weber, Susan (VHACLE)'" , Paul Bradbury
, karen adams ,
HistoNet
Server
Message-ID:
<03E1F5968F60C5448635D49D38B283ED797B597F@SJMEMXMBS11.stjude.sjcrh.local
>
Content-Type: text/plain; charset="us-ascii"
We get our B-plus from BBC Biomedical.
Charlene
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weber,
Susan (VHACLE)
Sent: Wednesday, May 07, 2008 12:10 PM
To: Paul Bradbury; karen adams; HistoNet Server
Subject: RE: [Histonet] B-plus question. .
I may have missed this, but may I have the name of the Vendor you
purchase your B-plus from. Thanks.
Susan M Weber HT(ASCP)
Histology Supervisor
Louis Stokes Cleveland VA Medical Center
10701 East Blvd
Cleveland, Ohio 44106
(216) 791-3800 X6154
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paul
Bradbury
Sent: Tuesday, May 06, 2008 9:53 AM
To: karen adams; HistoNet Server
Subject: Re: [Histonet] B-plus question
Hi Karen,
I have been using the same sequence of reagents for several years with
great success. We routinely fix bone marrow cores for 3 hours in B-plus,
rinse in water, decalcify, rinse again, and put the cassette in with all
the other tissues for processing. B-plus contains formaldehyde anyway,
so your are not introducing a different reagent when you transfer them
to formalin during processing.
B-plus gives much better cytological detail than formalin. Nuclear
detail is crisper, granules are better preserved, hemosiderin is not
effected. CD-3, CD-20, kappa and lambda, etc. all work beautifully after
B-plus. It also has the advantage of not producing an fixation artefact
pigment like B-5 does. I think you will be happy with your decision to
change.
Paul Bradbury
Kamloops, Canada
karen adams wrote:
> Hello all....we are changing bone marrow fixative from formalin to
B-plus.
> If we fix for the required time in B-plu and then process on the VIP
w/ the
> other specimens using formalin are there any effects on the tissue
going
> from B-plus to formalin??
> Thank you in advance
>
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 10
Date: Wed, 07 May 2008 15:02:31 -0500
From: "Shelly Christenson"
Subject: [Histonet] Immunohistotechnologist position open at Kansas
State Univ.
To:
Message-ID: <4821C486.EF61.003F.0@vet.k-state.edu>
Content-Type: text/plain; charset=US-ASCII
There is a position in the Immunohistochemistry section of the Histology
Lab of the Kansas State Veterinary Diagnostic Laboratory. The
successful candidate will have a background in Immunohictochemeistry,
Immunofluorescence, and other aspects of histology . Supervisory
experience will be beneficial. A Bachelor of Science or equivalent
degree and a minimum of one year of laboratory experience is required.
This is just a short version of the job description for more information
can contact:
Dr. Brad DeBey, DVM, PhD
785/532-4481
debey@vet.k-state.edu
Also it posited online
https://www.da.ks.gov/ps/esummary/es-online/frmes1.asp
Requisition number is 160443
Feel free to contact me also.
Shelly Christenson HT(ASCP)
785/532-4464
christen@vet.k-state.edu
------------------------------
Message: 11
Date: Wed, 7 May 2008 13:03:32 -0700 (PDT)
From: Rene J Buesa
Subject: RE: [Histonet] Lot numbers. .
To: "Henry, Charlene" , "'Webb, Dorothy
L'"
,
"histonet@lists.utsouthwestern.edu"
Message-ID: <314908.74685.qm@web65707.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
The only lot numbers I kept track of were those of "critical reagents",
namely: any one dealing with any IHC procedure and the few staining
solutions bought commercially.
Ren J.
"Henry, Charlene" wrote:
---------------------------------
Be a better friend, newshound, and know-it-all with Yahoo! Mobile. Try
it now.
------------------------------
Message: 12
Date: Wed, 7 May 2008 16:08:49 -0400
From: "Bonner, Janet"
Subject: RE: [Histonet] Lot numbers
To: "Webb, Dorothy L" ,
histonet@lists.utsouthwestern.edu
Message-ID:
<5F31F38C96781A4FBE3196EBC22D47807F262B@fhosxchmb006.ADVENTISTCORP.NET>
Content-Type: text/plain; charset=iso-8859-1
We have this happen when the chuck is the temperature of the cryostat
before we put the OCT on it. The OCT freezes before it has a chance to
seep into the grooves. Try putting the OCT on the warm chuck just after
you put the chuck on the bar in the cryostat.
________________________________
From: histonet-bounces@lists.utsouthwestern.edu on behalf of Webb,
Dorothy L
Sent: Wed 5/7/2008 1:56 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Lot numbers
Does everyone keep track of each reagent lot number, including, water,
formalin, alcohols, etc. as it is opened and used? What about recycled
products? I am trying to come up with a logsheet and would appreciate
any help in this area!
Also, we have recently run into problems with our freezing compound
coming off of the chucks during frozen sectioning. Any suggestions
and/or what is the method of freezing tissue that everyone uses for
their cryostat sections? Thanks fellow histotechs for helping me with
these matters that I am coming up empty on!!!!!!!!!!!!!!!!!!!!!
Dorothy Webb, HT (ASCP)
Histology Technical Supervisor
Regions Hospital, Pathology Department
640 Jackson Street, Saint Paul, MN 55101-2595
Phone: 651-254-2962
Fax: 651-254-2741
Regions Hospital is part of the HealthPartners family of care
________________________________________
This e-mail and any files transmitted with it are confidential and are
intended solely for the use of the individual or entity to whom they are
addressed. If you are not the intended recipient or the individual
responsible for delivering the e-mail to the intended recipient, please
be advised that you have received this e-mail in error and that any use,
dissemination, forwarding, printing, or copying of this e-mail is
strictly prohibited.
If you have received this e-mail in error, please immediately notify the
HealthPartners Support Center by telephone at (952) 967-6600. You will
be reimbursed for reasonable costs incurred in notifying us.
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
==========================3D=========================3D====
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 an employee 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.
==========================3D=========================3D====
------------------------------
Message: 13
Date: Wed, 7 May 2008 13:13:04 -0700
From: "Wright, Clarissa B CIV"
Subject: RE: [Histonet] c-kit (CD117) rat and mouse tissue
To: ,
,
Message-ID:
<8072C1D1C2865C49B7F0A95FC47B7646AABF35@NMCSD-EX-VS-01.nmed.ds.med.navy.
mil>
Content-Type: text/plain; charset="us-ascii"
Michelle,
Try Lifespan Biosciences, they have several. I have not tried them, so I
can't help you with the protocol.
Kris
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
michelle.schwab-macdonald@novartis.com
Sent: Wednesday, May 07, 2008 10:57 AM
To: histonet@lists.utsouthwestern.edu;
histonet-bounces@lists.utsouthwestern.edu
Subject: [Histonet] c-kit (CD117) rat and mouse tissue
I am looking for a c-kit Ab that will stain FFPE rat or mouse tissue. I
have heard that Dako has one for rat. Is anyone using it and willing to
share protocol information or does anyone have an Ab suggestion?
Thanks in Advance!! :-)
Michelle Schwab-MacDonald HT(ASCP)
Novartis, Boston, MA
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------------------------------
Message: 14
Date: Wed, 07 May 2008 21:00:51 +0000
From: zodiac29@comcast.net
Subject: [Histonet] histonet@lists.utsouthwestern.edu
To: histonet@lists.utsouthwestern.edu
Message-ID:
<050720082100.4274.48221883000BDFE3000010B22216557996C7CD0C0E070B0196@co
mcast.net>
Content-Type: text/plain
Hello,
I have read somewhere that silver solutions should be stored in an
expolsive proof refrigerator. Is this true? Right know we just store
them in a regular house hold type fridge. Can anyone comment on this?
Thanks,
Jenny
------------------------------
Message: 15
Date: Wed, 7 May 2008 14:11:39 -0700
From: "Laurie Colbert"
Subject: [Histonet] glassware cleaners
To:
Message-ID:
<57BE698966D5C54EAE8612E8941D768302E38DDF@EXCHANGE3.huntingtonhospital.c
om>
Content-Type: text/plain; charset="us-ascii"
I have used a detergent called Contrad 70 for years to clean my
glassware, but it is no longer available from my vendor. What are
others using to clean iron solutions, muci stain, silver, etc off of
their glassware?
Laurie
------------------------------
Message: 16
Date: Thu, 8 May 2008 09:17:57 +1000
From: "Tony Henwood"
Subject: RE: More on [Histonet] Looking for detailed protocol to stain
for PASin liver sections (paraffin)
To: "Gayle Callis" , "Histonet"
Message-ID:
Content-Type: text/plain; charset="us-ascii"
I have found the following Modified PAS procedure especially useful for
cytology smears that have been fixed in ethanol:
Alcoholic PAS Stain
Mucins and glycogen are water soluble. Over-rinsing slides, especially
cytological smears, could result in excessive loss of these PAS
substances. The following variant of the PAS stain performs the
reactions in alcohol, thus decreasing the loss of these substances.
Solutions:
1. Alcoholic Schiff reagent
Basic Fuchsin 0.5g
Ethanol 80ml
Distilled Water 20ml
Hydrochloric acid 1ml
2. Alcoholic Periodic acid
Periodic acid 0.5g
95% ethanol 50ml
Procedure:
1. Fix smears in 95% ethanol or use methanol fixed air-dried
smears.
2. Place in alcoholic periodic acid 20min.
3. Wash in alcohol
4. Place in alcoholic Schiff's 20min.
5. Rinse slides in alcohol to remove excess dye.
6. Rinse slides in water.
7. Counterstain slides in Haematoxylin 2min.
8. Rinse slides in water.
9. Differentiate and Blue.
10. Dehydrate, clear and mount.
References:
Horobin and Kevill-Davis (1971) Stain Techn 46:53-58.
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle
Callis
Sent: Thursday, 8 May 2008 2:06 AM
To: Histonet
Subject: Fw: More on [Histonet] Looking for detailed protocol to stain
for PASin liver sections (paraffin)
Dr. Nasonkin was staining for glycogen (per my question) and via private
emailing, it was suggested he not use aqueous formalin or PFA fixation,
but
an alcoholic fixative to retain the glycogen in the liver, do diastase
digestion to prove it is glyogen, and also process tissues - starting
in
higher (100%) alcohols to help retain glycogen. Suggested fixatives
were
Carnoy, Gendre and alcoholic formalin.
If you have any other suggestions for him, please add to the list of to
do's
.
Gayle M. Callis
HTL/HT/MT(ASCP)
Bozeman MT
> ----- Original Message -----
> From: "Igor Nasonkin"
> To: "Gayle Callis"
> Cc:
> Sent: Tuesday, May 06, 2008 6:35 PM
> Subject: Re: [Histonet] Looking for detailed protocol to stain for PAS
> in liver sections (paraffin)
>
>
> Gayle,
> Yes, glycogen.
> Does it mean that if we fixed livers in paraformaldehyde prepared on
> PBS
> we
> lost glycogen? What if we fixed the whole liver, not section on a
slide?
> One
> cannot remove glycogen from the whole liver this way. Thank you for
this
> info,
> Igor
>
> Dr. Igor O. Nasonkin
> Research Fellow
> National Institutes of Health/NEI
> 9000 Rockville Pike, MSC 1864
> Bldg 10, Room 10B11
> Bethesda, MD 20892
> Tel: 301-443-7398 -work
> 617-388-4104 -cell
> Fax 301-480-1769
> email: nasonkini@mail.nih.gov
> http://www.nei.nih.gov/intramural/nnrl.asp
>
> On 5/6/08 8:08 PM, "Gayle Callis" wrote:
>
>> You did not say what you are trying to see in the liver? Glycogen?
Some
>> other PAS positive tissue component? If so,aqueous formalin
fixation
>> will remove the glyocgen. An alcoholic fixative helps retain
>> glycogen.
>>
>> Gayle M. Callis
>> HTL/HT/MT(ASCP)
>> Bozeman MT 59715
>>
>> ----- Original Message -----
>> From: "Igor Nasonkin"
>> To:
>> Sent: Tuesday, May 06, 2008 5:24 PM
>> Subject: [Histonet] Looking for detailed protocol to stain for PAS in
>> liver sections (paraffin)
>>
>>> I am looking for a detailed protocol for PAS staining in liver
>>> sections (paraffin-embedded). The 1st set of sections did not work,
>>> and we had no
>>> +control since we have not done it. These are 4-6wk mouse liver
>>> +sections
>>> embedded in paraffin; our lab is experienced in IHC on paraffin
>>> sections so these were done right. But PAS staining did not work.
>>> What could be main reasons? Any positive control we could use? Thank
>>> you in advance, Igor
_______________________________________________
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addressed. If you are not the intended recipient, please delete it and
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Views expressed in this message and any attachments are those of the
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------------------------------
Message: 17
Date: Thu, 8 May 2008 09:29:21 +1000
From: "Tony Henwood"
Subject: RE: More on [Histonet] Looking for detailed protocol to stain
for PASin liver sections (paraffin)
To: "Gayle Callis" , "Histonet"
Message-ID:
Content-Type: text/plain; charset="us-ascii"
Sorry,
I have tried it on methanol fixed frozen sections of liver and it works
quite well
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318
-----Original Message-----
From: Gayle Callis [mailto:gayle.callis@bresnan.net]
Sent: Thursday, 8 May 2008 9:24 AM
To: Tony Henwood
Subject: Re: More on [Histonet] Looking for detailed protocol to stain
for PASin liver sections (paraffin)
Tony,
What a delightful variation and I bet it would work on tissue sections
too.
Gayle Callis
----- Original Message -----
From: "Tony Henwood"
To: "Gayle Callis" ; "Histonet"
Sent: Wednesday, May 07, 2008 5:17 PM
Subject: RE: More on [Histonet] Looking for detailed protocol to stain
for
PASin liver sections (paraffin)
I have found the following Modified PAS procedure especially useful for
cytology smears that have been fixed in ethanol:
Alcoholic PAS Stain
Mucins and glycogen are water soluble. Over-rinsing slides, especially
cytological smears, could result in excessive loss of these PAS
substances. The following variant of the PAS stain performs the
reactions in alcohol, thus decreasing the loss of these substances.
Solutions:
1. Alcoholic Schiff reagent
Basic Fuchsin 0.5g
Ethanol 80ml
Distilled Water 20ml
Hydrochloric acid 1ml
2. Alcoholic Periodic acid
Periodic acid 0.5g
95% ethanol 50ml
Procedure:
1. Fix smears in 95% ethanol or use methanol fixed air-dried smears. 2.
Place in alcoholic periodic acid 20min. 3. Wash in alcohol 4. Place in
alcoholic Schiff's 20min. 5. Rinse slides in alcohol to remove excess
dye. 6. Rinse slides in water. 7. Counterstain slides in Haematoxylin
2min. 8. Rinse slides in water. 9. Differentiate and Blue. 10.
Dehydrate, clear and mount.
References:
Horobin and Kevill-Davis (1971) Stain Techn 46:53-58.
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory
Manager & Senior Scientist The Children's Hospital at Westmead, Locked
Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle
Callis
Sent: Thursday, 8 May 2008 2:06 AM
To: Histonet
Subject: Fw: More on [Histonet] Looking for detailed protocol to stain
for PASin liver sections (paraffin)
Dr. Nasonkin was staining for glycogen (per my question) and via private
emailing, it was suggested he not use aqueous formalin or PFA fixation,
but an alcoholic fixative to retain the glycogen in the liver, do
diastase digestion to prove it is glyogen, and also process tissues -
starting in
higher (100%) alcohols to help retain glycogen. Suggested fixatives
were
Carnoy, Gendre and alcoholic formalin.
If you have any other suggestions for him, please add to the list of to
do's .
Gayle M. Callis
HTL/HT/MT(ASCP)
Bozeman MT
> ----- Original Message -----
> From: "Igor Nasonkin"
> To: "Gayle Callis"
> Cc:
> Sent: Tuesday, May 06, 2008 6:35 PM
> Subject: Re: [Histonet] Looking for detailed protocol to stain for PAS
> in liver sections (paraffin)
>
>
> Gayle,
> Yes, glycogen.
> Does it mean that if we fixed livers in paraformaldehyde prepared on
> PBS we
> lost glycogen? What if we fixed the whole liver, not section on a
slide?
> One
> cannot remove glycogen from the whole liver this way. Thank you for
this
> info,
> Igor
>
> Dr. Igor O. Nasonkin
> Research Fellow
> National Institutes of Health/NEI
> 9000 Rockville Pike, MSC 1864
> Bldg 10, Room 10B11
> Bethesda, MD 20892
> Tel: 301-443-7398 -work
> 617-388-4104 -cell
> Fax 301-480-1769
> email: nasonkini@mail.nih.gov
> http://www.nei.nih.gov/intramural/nnrl.asp
>
> On 5/6/08 8:08 PM, "Gayle Callis" wrote:
>
>> You did not say what you are trying to see in the liver? Glycogen?
Some
>> other PAS positive tissue component? If so,aqueous formalin
fixation
>> will remove the glyocgen. An alcoholic fixative helps retain
>> glycogen.
>>
>> Gayle M. Callis
>> HTL/HT/MT(ASCP)
>> Bozeman MT 59715
>>
>> ----- Original Message -----
>> From: "Igor Nasonkin"
>> To:
>> Sent: Tuesday, May 06, 2008 5:24 PM
>> Subject: [Histonet] Looking for detailed protocol to stain for PAS in
>> liver sections (paraffin)
>>
>>> I am looking for a detailed protocol for PAS staining in liver
>>> sections (paraffin-embedded). The 1st set of sections did not work,
>>> and we had no
>>> +control since we have not done it. These are 4-6wk mouse liver
>>> +sections
>>> embedded in paraffin; our lab is experienced in IHC on paraffin
>>> sections so these were done right. But PAS staining did not work.
>>> What could be main reasons? Any positive control we could use? Thank
>>> you in advance, Igor
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
*********************************************************************
This email and any files transmitted with it are confidential and
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solely for the use of the individual or entity to whom they are
addressed.
If you are not the intended recipient, please delete it and notify the
sender.
Views expressed in this message and any attachments are those of the
individual sender, and are not necessarily the views of The Children's
Hospital at Westmead
This note also confirms that this email message has been
virus scanned and although no computer viruses were detected, The
Childrens
Hospital at Westmead accepts no liability for any consequential damage
resulting from email containing computer viruses.
**********************************************************************
*********************************************************************
This email and any files transmitted with 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, please delete it and
notify the sender.
Views expressed in this message and any attachments are those of the
individual sender, and are not necessarily the views of The Children's
Hospital at Westmead
This note also confirms that this email message has been
virus scanned and although no computer viruses were detected, The
Childrens Hospital at Westmead accepts no liability for any
consequential damage resulting from email containing computer viruses.
**********************************************************************
------------------------------
Message: 18
Date: Wed, 7 May 2008 17:11:11 -0700 (PDT)
From: Akemi Allison-Tacha
Subject: Re: [Histonet] Re; Decal Process From the Past
To: HistoNet Server , Paul Bradbury
Message-ID: <251486.25591.qm@web31304.mail.mud.yahoo.com>
Content-Type: text/plain; charset=us-ascii
Hi Paul,
I appreciate your courteous reply to the older methodologies, but as I
stated, this was just "throwing an additional tidbit into the mix". As
you are most likely aware, histology in the past was not considered an
exact science. Histotech's referred to formula's as recipes and the
results for the most part, depended on the phase on the moon and the
technical skill and knowledge of the individual. Most of the
histotech's I ran a crossed in the 60's, 70's, 80's and even the 90's
were OJT. There are histotech's that are still following method's that
were passed on to them years and years ago and aren't sure why.
This procedure was for larger bone specimens. We never put BM cores
into RDO, that solution was much too harsh for such a fragile specimen.
For BM cores, we used a gentle decal solution that had EDTA incorporated
into it. It usually only took a half hour and we never left it longer
than an hour. We rinsed the decalcified BM core in running tap water,
then placed it on the processor.
When I started doing technical support for a biotech company, I became
aware of the "old recipes" and variations in adhering to set procedures.
There were a number of histotech's that had NO idea why they were doing
a procedure, or how each chemical step impacted the final result. These
histotech's had no clue of what theory and practice was all about. It
was a wonderful opportunity to try to help my fellow histologists in any
way possible and gave me a opportunity to grow as well. A day does not
go by that I don't grow and learn something new.
Akemi Allison-Tacha, BS, HT(ASCP)HTL
Client Services Manager
PhenoPath laboratories
551 North 34th Street, Suite 100
Seattle, WA 98103-8675
Work: (206) 374-9000 ext 1053
E-Mail: akemiat3377@yahoo.com
--- On Tue, 5/6/08, Paul Bradbury wrote:
> From: Paul Bradbury
> Subject: [Histonet] Re; RINSING IN TAP WATER AFTER DECAL
> To: "HistoNet Server"
> Date: Tuesday, May 6, 2008, 11:40 PM
> With all due respect to Akemi, the practice of
> "neutralizing" the
> decalcifying acid by treating the tissue with saturated
> sodium
> bicarbonate is not a good idea.
> In her e-mail she states that "The tissue fizzes a bit
> ..." no surprise
> there. Trying to retain good morphology in bone biopsy core
> is difficult
> enough without disrupting the tissues with bubbles of
> carbon dioxide. A
> bone core is usually only 2-3 mm in diameter, so washing
> out the
> decalcifying fluid will take only a few minutes. Once the
> acid has been
> removed, decalcification will stop.
> Paul Bradbury
> Kamloops, Canada
>
> ******************************************************************
>
> > Akemi Allison-Tacha wrote:
> >> Hi All,
> >>
> >> I am going to put an additional tidbit into the
> mix. I was instructed many many moons ago, (back in 1974)
> to 1st rinse off any residual RDO or decal solution with a
> brief rinse in tap water and then place the decal specimens
> into saturated sodium bicarbonate for 10 minutes to stop the
> decal process, otherwise the specimen will continue to
> decal. The tissue fizzes a bit, when this reaction ceases,
> then rinse in running tap water for an additional 10
> minutes.
> >>
> >> Akemi Allison-Tacha, BS, HT(ASCP)HTL
> >> Client Services Manager
> >> PhenoPath laboratories
> >> 551 North 34th Street, Suite 100
> >> Seattle, WA 98103-8675
> >> Work: (206) 374-9000 ext 1053
> >> E-Mail: akemiat3377@yahoo.com
> >>
> >>
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 19
Date: Wed, 7 May 2008 20:20:36 -0400
From: "jstaruk"
Subject: RE: [Histonet] Too many Techs...
To: "'Bonner, Janet'" , "'Anne van
Binsbergen'" , "'barbara carter'"
,
,
"'Histonet'"
Message-ID:
Content-Type: text/plain; charset="us-ascii"
_____________________
Jim Staruk
Mass Histology Service
www.masshistology.com
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner,
Janet
Sent: Wednesday, May 07, 2008 1:06 PM
To: Anne van Binsbergen; barbara carter;
histonet-request@lists.utsouthwestern.edu; Histonet
Subject: [Histonet] Too many Techs...
10,000cases? 3300 cases per tech per year, 100 cases per tech per
day?
At least 300 blocks min. per day per tech.
CAP has guidelines for justifying positions. I would also have the
techs
(like they need something else to do) keep track of their time for a
month.
It sounds as if three techs would minimally handle your lab. Not to
mention
two months worth of vacation/sick time taken per year.
Also - what is the boss seeing? Coffee breaks? Long lunch breaks?
We have 22 Techs, three shifts, six days per week, 50,000 Surgical
cases. It takes two techs worth just to cover time off per year. We do
not
attend procedures and we buy our reagents.
Good luck.
________________________________
From: histonet-bounces@lists.utsouthwestern.edu on behalf of Anne van
Binsbergen
Sent: Wed 5/7/2008 12:09 PM
To: barbara carter; histonet-request@lists.utsouthwestern.edu; Histonet
Subject: Re: [Histonet] Histology Coordinator Position in Kenosha
Wisconsin
i have a few questions for you if i may.
you have 3 techs serving 3 paths and 10 000 a year
apart from the basic routine histo operations, do your 3 techs
- do any grossing
- attend at renal biopsy collection
- do frozen sections
- take macro photos
- do slide and block filing
- source cases for sendout (consults/reviews) or do any archive
searches
- wash glassware,
- do specimen discards,
- make up formalin
i am facing pressure from my (non-histo) boss who is comparing my lab
and
staff to 'other modern facilities' and he now tells me i need to cut my
staff in half!!
i need ammo and numbers from others to build my defence
cheers
Annie
2008/5/7 barbara carter :
>
> I am leaving my position as Coordinator at United Hospital System and
am
> moving out of state and trying to help them recruit a new coordinator
to
> take my place.
>
> There is a position open at United Hospital System at the Kenosha
Campus
> in Kenosha Wisconsin for a Histology Coordinator. The hospital
receives
> about 10,000 surgical specimen a year with 3 Pathologists and 3
Histologist.
>
> Anyone interested please contact Sue Wergin Laboratory Supervisor at
> 262-656-5603.
>
>
>
>
> _________________________________________________________________
> Windows Live SkyDrive lets you share files with faraway friends.
>
>
http://www.windowslive.com/skydrive/overview.html?ocid=TXT_TAGLM_WL_Refr
esh_
skydrive_052008_______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
--
Anne (van Binsbergen) Hope
Abu Dhabi
UAE
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------------------------------
Message: 20
Date: Wed, 7 May 2008 20:27:40 -0400
From: "Amos Brooks"
Subject: [Histonet] Polymer IHC Detection
To: histonet@lists.utsouthwestern.edu
Message-ID:
<582736990805071727n10f00e42w643fadfd7f7b8d0f@mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Hi,
Does anyone know of a good polymer immunohistochemical detection kit
that labels antibodise raised in animals other than mouse or rabbit? I
am
looking for a goat or rat detection kit. I know I could use a mouse anti
goat then a mouse secondary, but I'd like to simplify the process. Of
course
vendor responses are more than welcome.
Thanks,
Amos Brooks
------------------------------
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End of Histonet Digest, Vol 54, Issue 9
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