[Histonet] Cytochrome Oxidase Stain
| From: | "Featherstone, Annette" |
PROCEDURE:
Fixation: frozen tissue
Technique: Unfixed frozen section at 10 microns thick.
Quality Control: Muscle tissue
Solutions:
Reagents:
1. 0.1 M Sodium Acetate buffer pH 5.6
Sodium Acetate Trihydrate--------------------------1.360g
Distilled water------------------------------------------100ml
Adjust pH to 5.6 with acetic acid. Store in refrigerator.
2. 1% Manganese Chloride
Manganese chloride------------------------------------1.0g
Distilled water-------------------------------------------100ml
3. 0.1% Hydrogen Peroxide
30% Hydrogen peroxide---------------------------------0.1ml (50ul)
Distilled water---------------------------------------------29.9ml (14.95)
Mix just before use.
4. 1% Cupric Sulfate
Cupric Sulfate---------------------------------------------1g
Distilled water---------------------------------------------100ml
5. 1.0 N Sodium Hydroxide
Sodium Hydroxide--------------------------------------4g
Distilled water-------------------------------------------100ml
6. Incubating medium
DAB-------------------------------------------------------0.03g
0.1 M sodium acetate buffer--------------------------13.5ml
1% manganese chloride--------------------------------1.5ml
0.1% hydrogen peroxide-------------------------------0.15ml
Mix together just before use. Adjust pH to 5.5 with NaOH. Filter before use.
Procedure:
1. Cut frozen sections at 10 microns.
2. Pour incubating solution over slides. Cover to prevent evaporation.
3. Incubate in 37°C oven for 1 hour.
4. Rinse in two changes distilled water, 5-10 seconds each.
5. Place in 1% cupric sulfate for 5 minutes.
6. Rinse in distilled water, 2-3 changes, 30 seconds total.
7. Dehydrate through graded alcohols, clear in xylene and coverslip.
Results:
Mitochondria------------------------------Brown
Type I fibers-------------------------------Darker brown
Type II fibers------------------------------Lighter brown
Myofibrils----------------------------------Unstained
DISTRIBUTION:
This policy shall be distributed to all pathologists, technical and secretarial staff of the Department of Pathology and Laboratory Medicine.
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of
histonet-request@lists.utsouthwestern.edu
Sent: Thursday, March 23, 2006 11:14
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 28, Issue 37
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Today's Topics:
1. RE: CAP - ANP.11713 Phase II (Weems, Joyce)
2. guidelines for Her2 i nterpretation (Goodwin, Diana)
3. RE: CAP - ANP.11713 Phase II (Horn, Hazel V)
4. The MT thing (Orr, Rebecca)
5. DAB in tablet form (meint002)
6. Fixation sensitive IHC markers (Thomas Crowell)
7. RE: Fixation sensitive IHC markers (Sebree Linda A.)
8. RE: CAP - ANP.11713 Phase II (Sebree Linda A.)
9. Re: DAB in tablet form (Andrea T. Hooper)
10. Re: guidelines for Her2 interpretation (Patti Loykasek)
11. p16 antibody (Dick Paulson [Source Medical Products])
12. Contaminated Alcohols from VIP processors (bamoe@gundluth.org)
13. Avoiding Re: [Histonet] DAB in tablet form (Gayle Callis)
14. Re: DAB in tablet form (Rene J Buesa)
15. RE: Contaminated Alcohols from VIP processors (Charles.Embrey)
16. CAP Standard ANP.11713 - Phase II (Jeff Silverman)
17. RE: Fixation sensitive IHC markers (Luis Chiriboga)
18. RE: Contaminated Alcohols from VIP processors (Monfils, Paul)
19. National Search for Histology Supervisor at the National
Cancer Institute; up to $84,559 per year
(Nellis, Kevin Lee (NIH/NCI) [E])
20. Re: Training Med Techs - some candid comments (Jennifer MacDonald)
21. MAK 6 HELP PLEASE!!!!!!!!!!! (Jesus Ellin)
22. Fw: Thermo/ Leica (MattG)
23. RE: Contaminated Alcohols from VIP processors (Bonner, Janet)
24. RE:JOB DESCRIPTION FOR HISTOLOGY ASSISTANT (mprice26@juno.com)
----------------------------------------------------------------------
Message: 1
Date: Wed, 22 Mar 2006 12:37:15 -0500
From: "Weems, Joyce"
Subject: RE: [Histonet] CAP - ANP.11713 Phase II
To: "Demarinis, Carolyn" ,
Message-ID:
<83AACDB0810528418AA106F9AE9B7F7E01D34E34@sjhaexc02.sjha.org>
Content-Type: text/plain; charset="US-ASCII"
We give the control to the pathologist daily - who initials
satisfactory or not... problems are dealt with by techs usually before
path gets here tho.
Joyce
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Demarinis, Carolyn
Sent: Wednesday, March 22, 2006 11:34 AM
To: HISTONET@PATHOLOGY.SWMED.EDU
Subject: [Histonet] CAP - ANP.11713 Phase II
There is a new question on the CAP checklist:
ANP.11713 - Phase II
"Is there documented evidence of daily review of technical quality of
histologic preparations by the pathologist?"
Our present procedure is the supervisor/technologist documents daily
review, but it is very clear that CAP is requiring the pathologist
to initial a document daily proving they have reviewed the technical
quality of slides.
How are other hospitals satisfying this requirement? It seems crazy to
make a pathologist initial a document daily,
when it is evident they review hundreds of slides a day, and they would
certainly report a problem to the supervisor when the stain is
unacceptable.
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information 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. Saint Joseph's Health System, Inc.
------------------------------
Message: 2
Date: Wed, 22 Mar 2006 12:46:10 -0500
From: "Goodwin, Diana"
Subject: [Histonet] guidelines for Her2 i nterpretation
To:
Message-ID:
<80CDD9C3FEEAFD4982B114C4A6DFD00EE4A3D4@uphsmbx2.UPHS.PENNHEALTH.PRV>
Content-Type: text/plain; charset="US-ASCII"
I have been out of the loop with Her- 2 for a while, and the last I
knew, it was left up to the individual labs to establish standards among
the interpreting physicians for interpretation, pretty much based on
the original Herceptest guidelines. Have any other standards been
established? If so, would someone please provide me with a reference?
Much obliged,
Diana Goodwin
Supervisor, Anatomic Pathology
Pennsylvania Hospital
Preston 655-C
ph. 215-829-6532
fax 215-829-7564
e-mail goodwind@[pahosp.com
------------------------------
Message: 3
Date: Wed, 22 Mar 2006 11:54:56 -0600
From: "Horn, Hazel V"
Subject: RE: [Histonet] CAP - ANP.11713 Phase II
To: "Demarinis, Carolyn" ,
HISTONET@PATHOLOGY.SWMED.EDU
Message-ID:
<9AE8AA9E1F644B4AA6C155FB6FD51C63038BDFFC@EMAIL.archildrens.org>
Content-Type: text/plain; charset=us-ascii
We send out a QC sheet daily with slides for the day. The docs sign
off on this sheet noting the quality of the slides and return it to our
lab. On the sheet it is just a matter of them checking off the slide
review and if they are acceptable or not.
HH
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Demarinis, Carolyn
Sent: Wednesday, March 22, 2006 10:34 AM
To: HISTONET@PATHOLOGY.SWMED.EDU
Subject: [Histonet] CAP - ANP.11713 Phase II
There is a new question on the CAP checklist:
ANP.11713 - Phase II
"Is there documented evidence of daily review of technical quality of
histologic preparations by the pathologist?"
Our present procedure is the supervisor/technologist documents daily
review, but it is very clear that CAP is requiring the pathologist
to initial a document daily proving they have reviewed the technical
quality of slides.
How are other hospitals satisfying this requirement? It seems crazy to
make a pathologist initial a document daily,
when it is evident they review hundreds of slides a day, and they would
certainly report a problem to the supervisor when the stain is
unacceptable.
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------------------------------------------------------
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 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.
==========================3D=========================3D=========================3D==
------------------------------
Message: 4
Date: Wed, 22 Mar 2006 12:33:00 -0600
From: "Orr, Rebecca"
Subject: [Histonet] The MT thing
To:
Message-ID:
Content-Type: text/plain; charset="US-ASCII"
AW I just love you guys!
Everyone has good points and I appreciate Mike's input.
The thing is I still don't have a supervisor and
I'll be very sad to leave my cushy IHC queendom to go do it myself.
Let's be done with this and move on.
Becky Orr CLA,HT(ASCP)
IHC Lead
Evanston Northwestern Healthcare
847-570-2771
------------------------------
Message: 5
Date: Wed, 22 Mar 2006 13:19:26 CST
From: meint002
Subject: [Histonet] DAB in tablet form
To: histonet@lists.utsouthwestern.edu
Message-ID: <200603221919.k2MJJQ8t021230@vanguard.software.umn.edu>
Content-Type: TEXT/plain; CHARSET=US-ASCII
Dear Histos,
Due to the hazards of working with 3,3' diaminobenzidine tetrahydrochloride
(DAB) for muscle enzyme staining, I am considering purchasing DAB as either
5mg, or 10mg tablets. Some companies have them dissolve in water and others
in a buffer solution.
I am looking for input from anyone who has used these tablets. Do you like
them, do they work well, staining results, which company, etc.... I know
that you can also get DAB in sealed separate vials, but it is much more
expensive and we are in an academic research setting where cost is a
consideration.
I am particularly interested in using them in the Cytochrome Oxidase stain.
I have tried many procedures for this stain and my Dr. is not all that
pleased with the results. If anyone has a good procedure for this stain, I
would be grateful if you would send me a copy.
I am revamping this lab and have had a lot of questions for the histonet that
you have been very helpful in addressing.
Thanks for all the suggestions in advance.
Joyce Meints
Histologist
University of Minnesota
Paul and Sheila Wellstone Muscular Dystrophy Center
MMC 206
420 Washington Ave. SE
Minneapolis, MN 55455-0392
e-mail: meint002@umn.edu
Fax: 612-625-8488
lab phone: 612-626-4703
------------------------------
Message: 6
Date: Wed, 22 Mar 2006 14:40:35 -0500
From: Thomas Crowell
Subject: [Histonet] Fixation sensitive IHC markers
To: histonet@lists.utsouthwestern.edu
Message-ID:
Content-Type: text/plain; charset="US-ASCII"
Dear Histonetters,
Can anyone suggest an antibody that I could use on human FFPE samples that
shows relative degrees of reactivity, based on variable factors such as
fixation time or tissue autolysis? I would like to assess the reactivity=20
of commercially available tissue bank samples.
Any suggestions would be appreciated.
Tom Crowell
Biogen Idec
Cambridge, MA
------------------------------
Message: 7
Date: Wed, 22 Mar 2006 14:00:54 -0600
From: "Sebree Linda A."
Subject: RE: [Histonet] Fixation sensitive IHC markers
To: "Thomas Crowell" ,
Message-ID:
Content-Type: text/plain; charset="US-ASCII"
Our pathologists have relied on a vimentin stain to assess the quality
of fixation and processing.
Linda Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-3224
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX: (608)262-7174
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Thomas
Crowell
Sent: Wednesday, March 22, 2006 1:41 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Fixation sensitive IHC markers
Dear Histonetters,
Can anyone suggest an antibody that I could use on human FFPE samples
that
shows relative degrees of reactivity, based on variable factors such as
fixation time or tissue autolysis? I would like to assess the
reactivity
of commercially available tissue bank samples.
Any suggestions would be appreciated.
Tom Crowell
Biogen Idec
Cambridge, MA
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 8
Date: Wed, 22 Mar 2006 14:10:49 -0600
From: "Sebree Linda A."
Subject: RE: [Histonet] CAP - ANP.11713 Phase II
To: "Rebecca Barnhart" ,
Message-ID:
Content-Type: text/plain; charset="US-ASCII"
In our lab (IHC & ISH), every case that goes out to the pathologist is
accompanied by the original request form that has a "QA" form on the
back. The pathologist is obligated to select "satisfactory" or
"unsatisfactory" in response to the statement: "I have reviewed the
patient and positive/negative control slides associated with the
immunostains requested on this case and find them
satisfactory/unsatisfactory" and initial his/her response.
Linda Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
A4/204-3224
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX: (608)262-7174
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca
Barnhart
Sent: Wednesday, March 22, 2006 11:44 AM
To: histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] CAP - ANP.11713 Phase II
I created a database that we enter our daily count (the date, first and
last case number, the number of blocks and slides). Also there are 2
prompts to be checked by the pathologist, one for H & E and the other
for cytology stain. We run a control as the first and last slide
everyday. If the stain is ok he puts a check in both boxes, if there is
a problem there is a spot to enter comments. I started doing it this
way so that if there is a problem with the stain I can easily go back
and see what cases it affected. I have a report that I can print to
show the quality of stain if an inspector request it. Previously we had
a sheet the pathologist kept at his desk and check each day if the stain
was ok and make comments if not.
I also use the spot for comments to document if we have changed
something (trying a different stain, new paraffin, added time to a step
on the processor or stainer, etc). So if there are questions or we need
to compare slides (because of a new product or new procedure) I have a
report that I run and I can easily pull the slides.
I also use this database to enter all statistical information. So at
any time I can give the pathologist or manager any number for anatomic
pathology.
>>> "Demarinis, Carolyn" 3/22/2006
11:34:14 AM >>>
There is a new question on the CAP checklist:
ANP.11713 - Phase II
"Is there documented evidence of daily review of technical quality of
histologic preparations by the pathologist?"
Our present procedure is the supervisor/technologist documents daily
review, but it is very clear that CAP is requiring the pathologist
to initial a document daily proving they have reviewed the technical
quality of slides.
How are other hospitals satisfying this requirement? It seems crazy
to
make a pathologist initial a document daily,
when it is evident they review hundreds of slides a day, and they
would
certainly report a problem to the supervisor when the stain is
unacceptable.
_______________________________________________
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: 9
Date: Wed, 22 Mar 2006 15:41:08 -0500
From: "Andrea T. Hooper"
Subject: Re: [Histonet] DAB in tablet form
To: Histonet , meint002
Message-ID:
Content-Type: text/plain; charset=iso-8859-1; format=flowed
I too am in academic labs so relate to the cost
concern; however, I strongly recommend DAB in
liquid kit form - either DAB+ from DAKO or from
Zymed. They are liquid ready to use concentrates
diluted into buffer either supplied with the kit
or you provide yourself.
I have used both for years and years now with
excellent results. Worth the extra bucks (about
80-100 dollars for 110 mls which is actually
quite a lot) for consistent results time after
time after time - without the hazards of working
with powder or solids.
Good luck!
Andrea
>Dear Histos,
>
>Due to the hazards of working with 3,3í diaminobenzidine tetrahydrochloride
>(DAB) for muscle enzyme staining, I am considering purchasing DAB as either
>5mg, or 10mg tablets. Some companies have them dissolve in water and others
>in a buffer solution.
>
>I am looking for input from anyone who has used these tablets. Do you like
>them, do they work well, staining results, which company, etc.... I know
>that you can also get DAB in sealed separate vials, but it is much more
>expensive and we are in an academic research setting where cost is a
>consideration.
>
>I am particularly interested in using them in the Cytochrome Oxidase stain.
>I have tried many procedures for this stain and my Dr. is not all that
>pleased with the results. If anyone has a good procedure for this stain, I
>would be grateful if you would send me a copy.
>
>I am revamping this lab and have had a lot of questions for the histonet that
>you have been very helpful in addressing.
>
>Thanks for all the suggestions in advance.
>
>
>Joyce Meints
>Histologist
>
>University of Minnesota
>Paul and Sheila Wellstone Muscular Dystrophy Center
>MMC 206
>420 Washington Ave. SE
>Minneapolis, MN 55455-0392
>
>e-mail: meint002@umn.edu
>Fax: 612-625-8488
>lab phone: 612-626-4703
--
------------------------------
Message: 10
Date: Wed, 22 Mar 2006 12:45:12 -0800
From: Patti Loykasek
Subject: Re: [Histonet] guidelines for Her2 interpretation
To: "Goodwin, Diana" , histonet
Message-ID:
Content-Type: text/plain; charset="US-ASCII"
Diana, yes the standard is the Herceptest guideline, as far as I know. I
haven't looked at some of the more recent FDA approved tests.
I highly recommend correlating your scores with Her2 FISH results. Also, if
you have >1 pathologist, the pathologists should periodically review cases
together so that their eyes stay "calibrated" to the same level of
staining/scoring.
Patti Loykasek BS, HTL, QIHC
PhenoPath Laboratories
Seattle, WA
I have been out of the loop with Her- 2 for a while, and the last I
> knew, it was left up to the individual labs to establish standards among
> the interpreting physicians for interpretation, pretty much based on
> the original Herceptest guidelines. Have any other standards been
> established? If so, would someone please provide me with a reference?
>
> Much obliged,
> Diana Goodwin
> Supervisor, Anatomic Pathology
> Pennsylvania Hospital
> Preston 655-C
> ph. 215-829-6532
> fax 215-829-7564
> e-mail goodwind@[pahosp.com
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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you are not the intended recipient, please contact the sender by e-mail
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------------------------------
Message: 11
Date: Wed, 22 Mar 2006 14:03:34 -0600
From: "Dick Paulson [Source Medical Products]"
Subject: [Histonet] p16 antibody
To:
Message-ID:
Content-Type: text/plain; charset="iso-8859-1"
Dr. Cartun,
Diagnostic BioSystems can provide the p16 antibody.
Clone(JC8) Isotype(IgG2a)
If you have any questions, please contact me directly.
Source Medical Products
Dick Paulson
dpconsult@earthlink.net
Phone (800) 393-6345
---- Original Message ----
We have been using Novocastra's p16 antibody (distributed by Vision
BioSystems here in the USA). I have been told that their antibody is
back-ordered. I know DAKO sells a p16 kit, but I'm only interested in
getting the antibody. Are there any other sources for p16 that work in
formalin-fixed, paraffin-embedded tissues? Thank you.
Richard
Richard W. Cartun, Ph.D.
Director, Immunopathology & Histology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT 06102
(860) 545-1596
(860) 545-0174 Fax
------------------------------
Message: 12
Date: Wed, 22 Mar 2006 14:58:31 -0600
From: bamoe@gundluth.org
Subject: [Histonet] Contaminated Alcohols from VIP processors
To: histonet@lists.utsouthwestern.edu
Message-ID:
Content-Type: text/plain; charset=US-ASCII
Hello all -
Has anyone had experience with contaminated alcohols coming off a
Tissue-Tek VIP processor?
We have two processors - one is a Miles VIP 3000, purchased in 1993, the
other is a Sakura VIP E300 purchased in 1996.
We have started experiencing the problem on both machines.
Our processing schedule utilizes 2 stations each of formalin, 80% ETOH,
recycled 95% ETOH, absolute ETOH (pure, not reagent grade), xylene, 3 or 4
paraffin stations, 1 cleaning xylene, 1 cleaning absolute ETOH.
The formalins and 80% are not contaminated, however the 95% and absolutes
show cloudiness when flushed with water down the drain. (We are assuming
that it's xylene causing this, but have not done analysis to confirm.)
Preventative maintenance was just done last week on both machines (by our
in-house biomed personnel) and we still have the cloudiness in the 95% and
absolute alcohols.
We have ruled out any problems with our recycler and know that the alcohols
are clean when they are put on the processor.
Any comments would be greatly appreciated!
Barb Moe
Gundersen Lutheran Medical Center
La Crosse WI
bamoe@gundluth.org
------------------------------
Message: 13
Date: Wed, 22 Mar 2006 14:27:28 -0700
From: Gayle Callis
Subject: Avoiding Re: [Histonet] DAB in tablet form
To: "Andrea T. Hooper" ,
Histonet@lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20060322142207.01b3db88@gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
I totally agree with Andrea. Another superb two part DAB substrate kit is
Pierce DAB (brown) $65 for 250 ml kit or Pierce CN/DAB (black) $78 for 250
ml kit, and you can make up as much or little as you want for number of
slides in a run.
Acadamic labs often get discounted pricing and free shipping other than
hazardous material costs, this can be purchased through Fisher and probably
VWR too.
At 01:41 PM 3/22/2006, you wrote:
>I too am in academic labs so relate to the cost concern; however, I
>strongly recommend DAB in liquid kit form - either DAB+ from DAKO or from
>Zymed. They are liquid ready to use concentrates diluted into buffer
>either supplied with the kit or you provide yourself.
>
>I have used both for years and years now with excellent results. Worth the
>extra bucks (about 80-100 dollars for 110 mls which is actually quite a
>lot) for consistent results time after time after time - without the
>hazards of working with powder or solids.
>
>Good luck!
>Andrea
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
406 994-4303 (FAX)
------------------------------
Message: 14
Date: Wed, 22 Mar 2006 13:30:00 -0800 (PST)
From: Rene J Buesa
Subject: Re: [Histonet] DAB in tablet form
To: meint002 , histonet@lists.utsouthwestern.edu
Message-ID: <20060322213000.38655.qmail@web61221.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Joyce:
I used for years the tablets with excellent results. I prefer the tablets and to dissolve them when needed.
René J.
meint002 wrote:
Dear Histos,
Due to the hazards of working with 3,3' diaminobenzidine tetrahydrochloride
(DAB) for muscle enzyme staining, I am considering purchasing DAB as either
5mg, or 10mg tablets. Some companies have them dissolve in water and others
in a buffer solution.
I am looking for input from anyone who has used these tablets. Do you like
them, do they work well, staining results, which company, etc.... I know
that you can also get DAB in sealed separate vials, but it is much more
expensive and we are in an academic research setting where cost is a
consideration.
I am particularly interested in using them in the Cytochrome Oxidase stain.
I have tried many procedures for this stain and my Dr. is not all that
pleased with the results. If anyone has a good procedure for this stain, I
would be grateful if you would send me a copy.
I am revamping this lab and have had a lot of questions for the histonet that
you have been very helpful in addressing.
Thanks for all the suggestions in advance.
Joyce Meints
Histologist
University of Minnesota
Paul and Sheila Wellstone Muscular Dystrophy Center
MMC 206
420 Washington Ave. SE
Minneapolis, MN 55455-0392
e-mail: meint002@umn.edu
Fax: 612-625-8488
lab phone: 612-626-4703
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
---------------------------------
Yahoo! Messenger with Voice. PC-to-Phone calls for ridiculously low rates=2E
------------------------------
Message: 15
Date: Wed, 22 Mar 2006 15:33:19 -0600
From: "Charles.Embrey"
Subject: RE: [Histonet] Contaminated Alcohols from VIP processors
To:
Cc: Histonet@pathology.swmed.edu
Message-ID:
Content-Type: text/plain; charset="us-ascii"
I had the same problem and worked with my sales rep. I was finally told
that what I was reporting was impossible. When it was verified by
equipment repair, my rep told me that the bottom line was that Sakura
sold me a tissue processor and it did process tissue. They made no
promise that I would be able to recycle the reagents so the matter was
closed. Needless to say, I replaced the VIP with a TBS unit and now
have no problem recycling my alcohol.
Charles Embrey PA(ASCP)
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
bamoe@gundluth.org
Sent: Wednesday, March 22, 2006 2:59 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Contaminated Alcohols from VIP processors
Hello all -
Has anyone had experience with contaminated alcohols coming off a
Tissue-Tek VIP processor?
We have two processors - one is a Miles VIP 3000, purchased in 1993, the
other is a Sakura VIP E300 purchased in 1996.
We have started experiencing the problem on both machines.
Our processing schedule utilizes 2 stations each of formalin, 80% ETOH,
recycled 95% ETOH, absolute ETOH (pure, not reagent grade), xylene, 3 or
4
paraffin stations, 1 cleaning xylene, 1 cleaning absolute ETOH.
The formalins and 80% are not contaminated, however the 95% and
absolutes
show cloudiness when flushed with water down the drain. (We are assuming
that it's xylene causing this, but have not done analysis to confirm.)
Preventative maintenance was just done last week on both machines (by
our
in-house biomed personnel) and we still have the cloudiness in the 95%
and
absolute alcohols.
We have ruled out any problems with our recycler and know that the
alcohols
are clean when they are put on the processor.
Any comments would be greatly appreciated!
Barb Moe
Gundersen Lutheran Medical Center
La Crosse WI
bamoe@gundluth.org
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Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 16
Date: Wed, 22 Mar 2006 17:08:34 -0500
From: Jeff Silverman
Subject: [Histonet] CAP Standard ANP.11713 - Phase II
To: cdemarinis@SARATOGACARE.ORG
Cc: histonet@lists.utsouthwestern.edu
Message-ID: <000001c64dfd$297e3740$6401a8c0@jeffrey028c8d9>
Content-Type: text/plain; charset=us-ascii
Previously, either myself of the lead tech for the day would examine a
control slide- a tonsil or fallopian tube from the day's workload, to
document proper staining and processing. To comply with this standard during
our December survey, we created a daily form (yet another) distributed to
whichever of our three pathologists is signing out that day. Any
deficiencies or problems in a specific slide including staining issues,
folds, wrinkles, or scratches, incomplete sections, misoriented tissues,
floaters or contaminants, bubbles under the coverslip, mislabeling are
listed by the docs. The histotech involved must address the problem and
document what was done to remedy the situation. This provides a tool for
continuing quality improvement so trends can be identified and individuals
with chronic problems can be retrained/counseled.
Jeffrey Silverman HT HTL QIHC (ASCP)
Pathologists' Assistant
Southside Hospital
Bay Shore New York USA
------------------------------
Message: 17
Date: Wed, 22 Mar 2006 18:02:36 -0500
From: Luis Chiriboga
Subject: RE: [Histonet] Fixation sensitive IHC markers
To: Thomas Crowell ,
histonet@lists.utsouthwestern.edu
Message-ID:
Content-Type: text/plain; charset=us-ascii
you might want to take a look at:
"Assessment of antigen damage in immunohistochemistry. the Vimentin internal
control" by Hector Battifora. American Journal of Clinical Pathology
November 1991 96(5) 669.
Hope it helps
Luis
____________________________________
Luis Chiriboga Ph.D.
NYU Cancer Institute and
Bellevue Hospital Center
New York University School Of Medicine
Department Of Pathology 4W27
462 First Avenue
New York, N.Y. 10016
W(212) 562-4667.
F(212) 263-2041
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Thomas
Crowell
Sent: Wednesday, March 22, 2006 2:41 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Fixation sensitive IHC markers
Dear Histonetters,
Can anyone suggest an antibody that I could use on human FFPE samples that
shows relative degrees of reactivity, based on variable factors such as
fixation time or tissue autolysis? I would like to assess the reactivity
of commercially available tissue bank samples.
Any suggestions would be appreciated.
Tom Crowell
Biogen Idec
Cambridge, MA
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------------------------------
Message: 18
Date: Wed, 22 Mar 2006 18:20:21 -0500
From: "Monfils, Paul"
Subject: RE: [Histonet] Contaminated Alcohols from VIP processors
To: "'histonet@lists.utsouthwestern.edu'"
Message-ID:
<09C945920A6B654199F7A58A1D7D1FDE017176AA@lsexch.lsmaster.lifespan.org>
Content-Type: text/plain; charset="iso-8859-1"
I have noticed the same thing, and I don't think the cloudiness is due to
xylene. I believe it is due to water-insoluble substances, particularly
lipids, which have leached out of the tissue samples into the stronger
alcohols. Because these substances are water-insoluble they don't tend to
dissolve in the formalin or the weaker alcohols where a lot of water is
present, but they do dissolve somewhat in the stronger alcohols. Also,
because they are water-insoluble, they precipitate (become cloudy) when
water is added to the alcohol, as in flushing down the drain.
Because I'm in a service research lab, I often run one kind of tissue per
processing run, and sometimes only one kind of tissue for several days. I
have noticed that the degree of cloudiness relates not only to the number of
specimens I have run since the last solvent change, but also very much to
the kind of tissues that were processed. I can run kidney or lung or
muscle specimens all week and there is minimal cloudiness when I discard the
alcohols. But if I process a large number of skin or breast or colon or
other fatty samples, the alcohol turns milky white when it hits the water=2E
> ----------
> From: histonet-bounces@lists.utsouthwestern.edu on behalf of
> bamoe@gundluth.org
> Sent: Wednesday, March 22, 2006 12:58 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Contaminated Alcohols from VIP processors
>
>
>
>
>
> Hello all -
>
> Has anyone had experience with contaminated alcohols coming off a
> Tissue-Tek VIP processor?
>
> We have two processors - one is a Miles VIP 3000, purchased in 1993, the
> other is a Sakura VIP E300 purchased in 1996.
> We have started experiencing the problem on both machines.
>
> Our processing schedule utilizes 2 stations each of formalin, 80% ETOH,
> recycled 95% ETOH, absolute ETOH (pure, not reagent grade), xylene, 3 or 4
> paraffin stations, 1 cleaning xylene, 1 cleaning absolute ETOH.
>
> The formalins and 80% are not contaminated, however the 95% and absolutes
> show cloudiness when flushed with water down the drain. (We are assuming
> that it's xylene causing this, but have not done analysis to confirm.)
>
> Preventative maintenance was just done last week on both machines (by our
> in-house biomed personnel) and we still have the cloudiness in the 95% and
> absolute alcohols.
>
> We have ruled out any problems with our recycler and know that the
> alcohols
> are clean when they are put on the processor.
>
> Any comments would be greatly appreciated!
>
> Barb Moe
> Gundersen Lutheran Medical Center
> La Crosse WI
> bamoe@gundluth.org
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
------------------------------
Message: 19
Date: Wed, 22 Mar 2006 18:25:57 -0500
From: "Nellis, Kevin Lee \(NIH/NCI\) [E]"
Subject: [Histonet] National Search for Histology Supervisor at the
National Cancer Institute; up to $84,559 per year
To:
Message-ID:
Content-Type: text/plain; charset="us-ascii"
Dear Collegues:
The NCI Laboratory of Pathology is are actively recruting for a
Histology Supervisor position. The job announcement is available on the
www.USAJobs.gov website. The position number is NCI-06-116352. You may
review the full details at:
http://jobsearch.usajobs.opm.gov/getjob.asp?JobID=41029650
There are three different government salary ranges for this position
based on the level of experience of the candidate. Please see the full
positing for details for examples of specialized experience. If a GS-10
level candidate is selected, the selected individual will be placed in a
Career Development Program. A successful candidate can be promoted to
the full level of a GS-12, during his/her career. The salary ranges
including locality pay are:
GS-10, Salary range $ 49,397 - $ 64,213
GS-11, Salary range $ 54,272 - $ 70,558
GS-12, Salary range $ 65,048 - $ 84,559
RELOCATION EXPENSES AND RECRUITMENT BONUS MAY BE PAID.
Interested parties may apply online at on the www.USAJobs.gov website or
you send an application directly to Ernesto Corrales. Be sure to read
the announcement carefully and supply all the information. Applicants
MUST submit a supplemental statement to their resume or application
addressing each of the specific KSAs. Only the most highly qualified
candidates will be referred to the hiring manager. The application
deadline is Tuesday, April 11, 2006.
For information about the NIH and surrounding areas, please see
www.nih.gov
For information about benefits and awards please see:
http://hr.od.nih.gov/default.htm
Thank you for your interest. Please forward this e-mail to anyone that
might consider this exciting opportunity.
Kevin L. Nellis, M.S., M.T. (A.S.C.P.)
Clinical Laboratory Manager
Laboratory of Pathology
Center for Cancer Research
National Cancer Institute
National Institutes of Health
U.S. Department of Health and Human Services
10 Center Drive, Room 2A33, MSC 1500
Bethesda, MD 20892
OFFICE: (301) 594-9532
FAX: (301) 402-0043
http://home.ncifcrf.gov/ccr/lop/Clinical/default.asp
------------------------------
Message: 20
Date: Wed, 22 Mar 2006 22:12:57 -0800
From: Jennifer MacDonald
Subject: Re: [Histonet] Training Med Techs - some candid comments
To: "Mike Kirby"
Cc: "Histonet \(E-mail\)"
Message-ID:
Content-Type: text/plain; charset="ISO-8859-1"
Dear Mr. Kirby,
I do not usually get involved in the mu but your response has changed that. Your Becky and to all Histotechs. Becky was not ins interpretation is that she is frustrated with Hi overlooked for management positions in favor of Med Techs.&n train a Med Tech with no histology experience if you have a very c apable Histotech available? She did not mean to imply that
Histotechn meant that o train somebody on t correct me if I am wrong.
As a Med Tech, I have worked in all areas of those fancy biochemistry and hematology analyzers, as matching blood for an ER patient=2E I know what stress i work in histology where things can also go wrong and depend the laboratory the turn-around-time expectations can be unreasonable . What we do in Histology can also impact a patient's health. A lthough the Pathologist makes the diagnosis, it is made on slides that
we h grievous pr your ability hands throughout
< gruelin Techs. Many h
There are many of us wh considered a boring "dead end division" field because we truly enjoy wh the respect we get from people like
Jennifer MacDonald
-----histonet-bounces@lists.utsouthwestern.edu wrote
To: "Histonet \(E-mail\)"
Fro Sent by: histonet-bounces@ Date: 03/22/2006 05:57AM
Subject: [Histonet] Dear Ms Orr.
You ma one!
Cli Your comment " Training a MT to be train a policeman to be a fireman" rather s I would like to follow with a counter
comment, "C can be shown how to the flames.
Granted, His requiring fine manual dext better than operating a high output Bi analyser or cross-matching a pint of blood, where a can kill a patient. Plus you operate under a fraction of the stress we are subjected to - try working for a full day, and then
doi expected to report f It's a gross insult to insinuate that ours is a "career".
As students, we spent 5 - 6 mo was available in the lab - Chempath, Ha Cyto, Immunology, Human genetics, blood transfusio even Histopath. When we passed our finals, we were allowed to c hoose which discipline we wanted to further our careers, and each
year, wit Histopath, as it was cons (Yes, pun intended)=2E
As fo Dept, then you c systems apply, regardless o practical applications of the work in hand Histopathology is just one of the services in the medi you don't walk on water, and neither do we. As far as I am conce rned, once trained as a general Med Tech, you can be trained in
virtually a thumbs" me, who after 35 half decent section or make a passa good as a manager!)
End of tirade - climb into bunker, to await the verbal unleashed................
Mr.M.Kirby
J South Africa
-----Original Message-----
Fr [[1]mailto:histonet-bounces@lists.utsou Of Orr, Rebecca
Sent: 17 March 2006 17:04
To Cc: &n Subject: [Histonet] Training Med Techs
H I would appreciate any feedback from those of you who may to train MT's (ASCP) to work in Histology.
They would be train them into Anatomic Patho Candid comments welcome, especial histology!
To me it would be like trying to fireman, it's a career, not a job, right?
We see a HT shortage in the Chicago area, but I am unsure how to
addre Degreed individuals have proven critical thinkin traditional education pathway, so I see the advantages, but ignore very capable HT managers with proven management and
organizationa an issue with me.I mean it's not like Non degreed HT's are stooopid
or something.
< Becky
Becky Orr CLA,HT(ASCP)
IHC Lead
Eva 847-570-2771
____ ______________________ 5F__ Histonet mailing Histonet@lists.utsouthwestern.edu
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[3]http://li
References
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3. 3D"http://lis=/
------------------------------
Message: 21
Date: Thu, 23 Mar 2006 05:34:29 -0700
From: "Jesus Ellin"
Subject: [Histonet] MAK 6 HELP PLEASE!!!!!!!!!!!
To: histonet@lists.utsouthwestern.edu
Message-ID:
Content-Type: text/plain; charset=us-ascii
Hello everyone I need some really big help we are trying to add an additional antibody to our system MAK6 from Zymed. We are using Ventana stainers, one is a Benchmark the other is LT. If anyone out there is using this antibody on these stainers and having success, PLEASE!!!!!!!!!!!! let me have your protocol for pariffin embeded and formalin fixed tissue.
Jesus Ellin
Yuma Regional Medical Center
------------------------------
Message: 22
Date: Thu, 23 Mar 2006 15:00:04 -0000
From: "MattG"
Subject: [Histonet] Fw: Thermo/ Leica
To:
Message-ID: <001501c64e8a$785f4240$6501a8c0@acer311vpbceh0>
Content-Type: text/plain; format=flowed; charset="iso-8859-1";
reply-type=response
Does anyone have a current email address for Pauline Fisher of Thermo
(formerly Shandon)?
I'd also be grateful if someone could send me an electronic version of the
user manual for the Leica TP1050.
If there are any UK based Leica or Thermo reps on here, could one of you
please drop me an email as I have a couple of questions I'd like to ask.
Thanks,
Matt Griffiths
___________________________________________________________
Win a BlackBerry device from O2 with Yahoo!. Enter now. http://www.yahoo.co.uk/blackberry
------------------------------
Message: 23
Date: Thu, 23 Mar 2006 10:34:15 -0500
From: "Bonner, Janet"
Subject: RE: [Histonet] Contaminated Alcohols from VIP processors
To: bamoe@gundluth.org, histonet@lists.utsouthwestern.edu
Message-ID:
Content-Type: text/plain; charset=iso-8859-1
I experienced this problem before and discovered the 95% ETOH was the culprit ( the 100%s could be white due to the 95% carried over into them). The company is going to deny this of course, but our supplier later came back to say there was a manufacturing problem that was not picked up on in their QC. Try another brand before you do anything, or call your supplier to ask if there are any other reported problems. -Janet
________________________________
From: histonet-bounces@lists.utsouthwestern.edu on behalf of bamoe@gundluth.org
Sent: Wed 3/22/2006 3:58 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Contaminated Alcohols from VIP processors
Hello all -
Has anyone had experience with contaminated alcohols coming off a
Tissue-Tek VIP processor?
We have two processors - one is a Miles VIP 3000, purchased in 1993, the
other is a Sakura VIP E300 purchased in 1996.
We have started experiencing the problem on both machines.
Our processing schedule utilizes 2 stations each of formalin, 80% ETOH,
recycled 95% ETOH, absolute ETOH (pure, not reagent grade), xylene, 3 or 4
paraffin stations, 1 cleaning xylene, 1 cleaning absolute ETOH.
The formalins and 80% are not contaminated, however the 95% and absolutes=20
show cloudiness when flushed with water down the drain. (We are assuming
that it's xylene causing this, but have not done analysis to confirm.)
Preventative maintenance was just done last week on both machines (by our=20
in-house biomed personnel) and we still have the cloudiness in the 95% and
absolute alcohols.
We have ruled out any problems with our recycler and know that the alcohols
are clean when they are put on the processor.
Any comments would be greatly appreciated!
Barb Moe
Gundersen Lutheran Medical Center
La Crosse WI
bamoe@gundluth.org
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 24
Date: Thu, 23 Mar 2006 16:11:00 GMT
From: "mprice26@juno.com"
Subject: [Histonet] RE:JOB DESCRIPTION FOR HISTOLOGY ASSISTANT
To: HISTONET@LISTS.UTSOUTHWESTERN.EDU
Message-ID: <20060323.081133.783.179@webmail01.nyc.untd.com>
Content-Type: text/plain
HI HISTONETTERS,
Can someone be so kind as to share a job description with me for a histology dept.assistant/aide (Non Registered).
Thank you.
Marsha Price
------------------------------
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End of Histonet Digest, Vol 28, Issue 37
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