Ouch!
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cartun, Richard
Sent: Thursday, April 30, 2015 4:34 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC billing question
Effective
The specific pages in the Dako Education Guide: Immunohistochemistry
Staining Methods, Fifth Edition are: Discussion on page 32 and
references on page 33. It's in the Fixation and Processing Chapter and
says no part of the process should have temperatures above 60C.
Rick Boen, BS, HTL
...@charter.net]
Sent: Sunday, April 26, 2015 10:42 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Re: IHC and Oven Temperatures
The specific pages in the Dako Education Guide: Immunohistochemistry
Staining Methods, Fifth Edition are: Discussion on page 32 and
references on page 33. It's
Thank you. I knew it was discussed in that reference. I guess my memory isn't
totally gone!
Joelle Weaver MAOM, HTL (ASCP) QIHC
Date: Sun, 26 Apr 2015 11:42:04 -0400
From: richardb...@charter.net
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Re: IHC and Oven
The95 for HIER is in liquid, The 82in the oven is dry slides. While wet high
temps enhance HIER, It seems the high dry temp does harm epitopes (there is a
paper somewhere on this but I don't have access to it right now).
Tim Morken
Supervisor, Histology, Electron Microscopy and Neuromuscular
Date: Thu, 16 Apr 2015 14:35:59 +
CC:
Subject: [Histonet] RE: IHC Billing Question
We have not been charging for the negative control, assuming that it was just
a cost of doing business. I would be interested to hear if anyone has been
charging for their negative controls as well
We recently added HER2 IHC testing in our lab which we are required to use a
negative reagent control
For each case. Is there a cpt code for negative reagent control reimbursement?
Any information on this
Would be much appreciated!
Thanks
Brandy Burnett
Histotechnoligist, QIHC(ASCP)
CCH
Moreira'; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: IHC Billing Question
We recently added HER2 IHC testing in our lab which we are required to use a
negative reagent control For each case. Is there a cpt code for negative
reagent control reimbursement? Any information on this Would
I have never heard of anyone billing for a negative control and would
highly recommend against doing so. As mentioned, it is a cost of doing
business--which is why when CAP clarified that polymer detection systems
did not require them, most cost-conscious labs quit running them.
Regards,
Bryan
Brandy Joana
Controls are never billable, a control test does not produce a usable result
based on the patient's specimen. The results of control tests - positive or
negative only tell you that your reagents, stains, antibodies etc. are
performing as expected.
| Dawn Hanson | Lab Manager |
Karla,
We try to run 1/2 of the required cases with known negative cases and the other
1/2 with known positive cases. In addition, all cases are run with a negative
reagent control in place of the antibody using the optimized protocol for the
antibody. These are your negative reagent
Our institution is billing IHC identical to what Joyce stated.
Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Holy Redeemer Hospital Laboratory
1648 Huntingdon Pike
Meadowbrook, PA 19046
Ph: 215-938-3676
Fax: 215-938-3874
Message: 3
Date: Wed, 8 Oct 2014 14:44:54 -0500
From: Horn, Hazel
I have used the CAP published 'Quality Management in Anatomic Pathology' and it
has been very helpful with this:
For a well-characterized antibody with a limited spectrum of antigenic
targets, like chromogranin, or prostate specific antigen, the validation can be
limited. A panel of 10
Hello Walter,
Many years ago I did Timm's Silver stain on free-floating rat brain slices.
The rats were perfused before removal and freezing of the brain on dry ice. I
then would cut slices in a cryostat and float the sections in saline until
ready to stain.
I would imagine that it might be
I had to revalidate all of my antibodies when we changed our IHC
platform. I used store bought TMAs and passed my CAP inspection with
flying colors. The cost savings is enormous. Ideally, they should be
processed the same as your test tissue, but since our CAP proficiency
slides are not, I did
Braud tbr...@holyredeemer.com
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إلى: histonet@lists.utsouthwestern.edu
الموضوع: [Histonet] RE: IHC Validation with TMAs
I had to revalidate all of my antibodies when we changed our IHC
platform. I used store bought TMAs and passed my CAP inspection
: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Terri Braud
Sent: Monday, August 04, 2014 1:30 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: IHC Validation with TMAs
I had to revalidate all of my antibodies when we changed
Thank you to everyone who sent a template. This is very helpful. I do
need clarfication on the 10 cases that are required for validation. Do you
include the 10 cases on this same form, or do you have something separate
for that? These look ideal for optimization. From what I have read
online,
I document all of the worksheet process¹ and then I create a ³Validation
Report² where my technical director both reviews all slides and signs this
report.
Michael Ann Jones, HT (ASCP)
Histology Manager
Metropath
7444 W. Alaska Dr. #250
Lakewood, CO 80226
303.634.2511
mjo...@metropath.com
On
Hi Kari,
I'd suggest taking advantage of CAP proficiency testing surveys so as to
compare your lab's results to others.
Linda A. Sebree
From: histonet-boun...@lists.utsouthwestern.edu
[histonet-boun...@lists.utsouthwestern.edu] on behalf of Delray Beach
The patient results are in the final report. The 2 year requirement is for QC
documents only.
Tim Morken
Supervisor,Hisotlogy, Electron Microscopy and Neuromuscular Special Studies
UC San Francisco Medical Center
San Francisco, CA
-Original Message-
From:
Allsion
It is my understanding, and I have gone over this with many billing experts as
well and performed endless research on. For medicare appropriate billing with
multiplex anitbodies, use the GO461 for first multiplex antibody then GO462 for
each separately identified antibody that the
The G codes are for Medicare/Medicaid. G0461 - the first and G0462 - each
additional per SPECIMEN.
88342 is per BLOCK. And you understand 88343...
Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.we...@emoryhealthcare.org
www.saintjosephsatlanta.org
5665 Peachtree
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: IHC antibody optimizing validating
CAP is very clear that in order to validate a new antibody, that: once the
stain has been optimized, that for a well characterized antibody with a limited
spectrum of antigenic targets, like
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: IHC antibody optimizing validating
CAP is very clear that in order to validate a new antibody, that: once
the stain has been optimized, that for a well characterized antibody
with a limited spectrum of antigenic targets, like Chromogranin
:
Subject: [Histonet] RE: IHC antibody optimizing validating
Fortunately, they say nothing at all because if that were the case, they
would no longer be able to peddle their Proficiency Programs for IHC,
since those too, are fixed and processed elsewhere.
Terri L. Braud, HT(ASCP)
Anatomic
Lester
I would not, first of all not all antibodies that work in human will cross
react with a particular species of animal. Another thing to consider is that
most human based detection systems consist of dual link reagents, meaning that
they work on both mouse and rabbit primaries these
@lists.utsouthwestern.edu
Subject: [Histonet] RE: IHC on animals
Lester
I would not, first of all not all antibodies that work in human will cross
react with a particular species of animal. Another thing to consider is
that most human based detection systems consist of dual link reagents,
meaning that they work
Hi Cassie,
I have tried going 'requisition free a number of times in at least three
institutions. The problems I've always encountered have been with the OR
nursing staff, who frankly tell us that they are too busy to have to do one
more thing - even if they were doing the 'thing' on paper
: [Histonet] RE: IHC antibody optimizing validating
Hi Cassie,
I have tried going 'requisition free a number of times in at least three
institutions. The problems I've always encountered have been with the OR
nursing staff, who frankly tell us that they are too busy to have to do one
more thing - even
CAP is very clear that in order to validate a new antibody, that: once
the stain has been optimized, that for a well characterized antibody
with a limited spectrum of antigenic targets, like Chromogranin or PSA,
the validation can be limited. A panel of 10 positive and 10 negative
neoplasms would
guidelines for
sure- where would that land me?
Joelle Weaver MAOM, HTL (ASCP) QIHC
Date: Tue, 25 Mar 2014 15:34:30 -0400
From: tbr...@holyredeemer.com
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: IHC antibody optimizing validating
CAP is very clear that in order
Are you changing the antibody dilution? If so, I would run 2-3 positive and
negative cases and make sure that the immunoreactivity compares favorably to
what you observed before.
Richard
Richard W. Cartun, MS, PhD
Director, Histology Immunopathology
Director, Biospecimen Collection Programs
On my IHC validation documents I have a space for comments. If I change
anything in a protocol I make a note in the comments space. I haven't had any
issue with any accrediting agencies.
Barbara S. Tibbs
Histology Supervisor
Accurate Diagnostic Labs
South Plainfield, NJ
Victoria
Cartilage can be tricky, proper processing and sectioning is important
initially. The samples can sometimes be cartilage only or both cartilage and
the underlying bone, we have worked with both. It also depends upon what
species you are working with. We have worked with mouse up to
I've used many different platforms and have liked most. Currently we
are using BioCare. We LOVE our BioCare Intellipath FLX for our IHC
platform. It is very cost effective, completely open, and has very user
friendly software. BioCare has been a super company to work with.
Fabulous customer
PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: IHC
I've used many different platforms and have liked most. Currently we are using
BioCare. We LOVE our BioCare Intellipath FLX for our IHC platform. It is very
cost effective, completely open, and has very user friendly software
Try ScyTek - The ready-made price is nearly as cheap as I can make it myself.
http://www.scytek.com/default.asp
Tresa
Tresa Goins
Histopathology Section Supervisor
Montana Veterinary Diagnostic Lab
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
Hey Tom,
Although we have a higher workload than you, ~ 1500 slides/month, we offer same
day TAT with a cut-off of noon. There are a couple exceptions, i.e. triple
stains and HER2 dual ISH but otherwise, utilizing 4 Ventana Ultras, we have it
all out by 4 pm or slightly later. Our hours are
We previously had two Ventana Benchmarks and was only able to get one set of
stains off per day. We had an 11 am cut-off which worked well. We would do
overnight stains as well so at least we were able to take off stains the
following morning. We replaced the Benchmarks with two Leica Bond
Hi Colleen,
We are just wrestling with this very issue and have decided to use:
Normal colon, i.e. staining present with all four antibodies
Colon carcinoma without the mutation, i.e. staining present with all
four antibodies
Colon carcinoma(s) with the mutation for each
] RE: IHC Tissue
Hi Colleen,
We are just wrestling with this very issue and have decided to use:
Normal colon, i.e. staining present with all four antibodies
Colon carcinoma without the mutation, i.e. staining present with
all four antibodies
Colon carcinoma(s
Linda,
we do it here
Ronnie Houston, MS HT(ASCP)QIHC
Anatomic Pathology Manager
ChildLab, a Division of Nationwide Children's Hospital
www.childlab.com
700 Children's Drive
Columbus, OH 43205
(P) 614-722-5450
(F) 614-722-2899
ronald.hous...@nationwidechildrens.org
www.NationwideChildrens.org
Vincent, you might want check out CLIA 88' complex testing requirement before
you write that job description.
Cassandra Davis
cda...@che-east.org
302-575-8095
Saint Francis Hospital
Saintfrancishealthcare.org
Saint Francis Facebook Page
From:
We just stain right over the HE. No problems.
Tom McNemar, HT(ASCP)
Histology Co-ordinator
Licking Memorial Health Systems
(740) 348-4163
(740) 348-4166
tmcne...@lmhealth.org
www.LMHealth.org
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
@lists.utsouthwestern.edu) histonet@lists.utsouthwestern.edu
Sent: Friday, March 15, 2013 1:05 PM
Subject: [Histonet] RE: IHC validation
Laurie, for first time validation use controls. Don't use patient cases that
are done for initial diagnostics. If they are repeats to test the system, that
is ok.
The question
Laurie, for first time validation use controls. Don't use patient cases that
are done for initial diagnostics. If they are repeats to test the system, that
is ok.
The question is, who tested the controls? Ideally you will send a
representative sample of your slides to another lab that uses the
Hello Dorothy,
Here we require HT or HTL certification for IHC positions. I look for
individuals that want to learn and have a desire to do IHC. I prefer folks
with experience, but since experienced techs are hard to pry away from other
employers, I am willing to train the right people. I
Hi Dorothy,
Take a quick look at the NSH website for job listings and maybe another
career's website like Simplyhired.com or Monster and get some idea as to how
the jobs for IHC lead techs are listed.
The problem with making absolute requirements (BS, HTL, QIHC, etc) is there are
a set of
All you need to do is compare the new clone to your current clone and show the
new clone is concordant with the new clone for sensitivity and specificity .
You don't need to compare to the same clone elsewhere.
Tim Morken
Department of Pathology
UC San Francisco Medical Center
505 Parnassus
Thank you all for the info. I agree that in-house validation is the way
to go. The only reason we had been using an outside reference lab was
due to recommendations during a JC inspection which were applied to our
area, but not really meant for IHC. The inspector was not a histology
person, so
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Dessoye,
Michael J
Sent: Friday, May 04, 2012 9:05 AM
To: Morken, Timothy; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: IHC Validation
Thank you
Dear Colleagues
What secondary antibody or polymer would be better to use for mouse primary
antibody on FFPE porcine tissues?Also, please share your experience regarding
macrophages detection in porcine arteries. I found couple in Abcam and Serotec
(only for frozen), but your opinions is
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Galina Deyneko
Sent: Monday, April 16, 2012 1:00 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Re: IHC in porcine tissues
Dear Colleagues
What
Consensus: Vimentin
Thanks' to all who responded!!
Luis Chiriboga Ph.D, Director
OCS Experimental Pathology IHC Core Lab
NYUSOM/Bellevue Hospital Center
Department of Pathology 4w27
(212) 562-4667
luis.chirib...@nyumc.org
From: Chiriboga, Luis
Sent: Tuesday, March 06, 2012 9:02 AM
To:
I am not sure if they can do that because CAP ANP.22570, explain this.
Tunde Ajibade BS, HTL(ASCP)QIHC
Histology Supervisor
Medical Center Hospital
Odessa,TX
Tel:432-640-2348
Fax:432-640-2303
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tunde Ajibade
Sent: Thursday, February 16, 2012 10:40 AM
To: 'histonet@lists.utsouthwestern.edu'
Cc: 'histonet-boun...@lists.utsouthwestern.edu'
Subject: [Histonet] RE: IHC and negative controls?
I am not sure if they can do
Thanks to everyone for the quick responses.
I am looking at ANP.22580 rev 12/12/2006 and it clearly talks about a separate
section of patient tissue... but just a little further it talks about using
internal negative controls (The type of negative tissue control used (i.e.,
separate sections,
Sara
If you can you want to stick with an enzyme digestion such as proteinase K or
another enzyme. If the antibody requires HIER and will not work with enzyme
digestion then modify your retreival time and temp, we use 70C for 2 hours and
find that this works for most antibodies. Some
I have place the deparaffinized slides into 10% neutral buffered formalin for
about 30 minutes before the heat retrieval. Somehow this seems to work
You can also lower the temperature of your retrieval and lengthen the time.
Good Luck.
Loralee McMahon, HTL (ASCP)
Immunohistochemistry
Tonsil is the control we use
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308-1592
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
www.premierlab.com
Ship to address:
1567 Skyway Drive, Unit E
Longmont, CO 80504
Treat the tissue exactly as if it was human.
Sure, your anti CD68 may not work on pig but, it's worth a try.
LBH? Probably LDH. If so, better to measure serum levels?
Trypen blue? Prob. Trypan blue. It's a dye used for cell viability tests.
So, it seems that 2 and 3 are not IHC at all.
More
Carl is correct... treat the tissue as if it is human... i.e., use
anti-mouse IgG as your link secondary Ab for CD68 mouse primary antibody,
etc. Not all CD68s cross-react with non-primates; I can let you know which
clones I've tried when I get back into work on Monday.
Jan Shivers
IHC
The three pathology services I've worked in in the last few yers that
did immunohistochemical staining for Helicobacter pylori did it on all
gastric biopsy specimens. Usually R/O H.p. is on the requisition.
I prefer a blue stain (Giemsa or Diff-Quik II or equivalent) if I have
one slide a day to
No, the best sample for negative control is NOT a section of your positive
control tissue.
The positive control would have already been tested for spurious
cross-reactions. Why continue to test it?
The patient's tissue has not been tested. So a mirror section is used.
Does the patient's tissue
We currently do. And we have the Dako system. So everytime we print the IHC
reports it takes pages and pages. The pathologist are supposed to sign off on
them and return them to me. I am then supposed to file them. But...
The major problems are
it takes up hundreds of sheets of paper
1) NO
2) Yes...at 4C
3) Sure, if somebody is around to do thisor you want to make somebody
go in and do it ;-)
carl
carl
Carl Hobbs
Histology Manager
Wolfson Centre for Age-Related Diseases
School of Biomedical Sciences
King's College London
Guys Campus
London SE1 1UL
Tel.020
Jana,
Since you are fixing in formaldehyde, why not paraffin embed them?
Frozen sectioning of formalin fixed tissue and doing IPX on them is difficult
as well as problems with storing the tissue.
I would cut frozen of unfixed tissue and do IPX on them rather than doing IPX
on formalin fixed
Dear Neil,
I was able to find a protocol like you described published where they had done
a regular IHC protocol on free-floating tissue sections for a-MSH antibody
using 0.3% H2O2, normal serum block, etc. They go on to describe that for
P-STAT3, the tissue needed to be pretreated with 1%
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Re: IHC pretreatment with NaOH/H2O2
Dear Neil,
I was able to find a protocol like you described published where they
had done a regular IHC protocol on free-floating tissue sections for
a-MSH antibody using 0.3% H2O2, normal serum block, etc
You did not say WHICH plastic?
Immunostaining on tissues embedded in methyl methacrylate are successful
since one can totally remove this plastic from the tissue with various
solvents. There may be some stringent antigen retrieval needed . Neil Hand
had a panel of over 200 antibodies
The Leica Bond detection kits are better than LSAB. They are barcoded for the
Bondmax machine-not sure if they offer packaged another way.
Joanne Mauger HT(ASCP)QIHC
Children's Hospital of Philadelphia
From: histonet-boun...@lists.utsouthwestern.edu
Sarah Goebel asked about prolonged incubation of primary antibodies on an
IHC.
You can do this, usually without increased background or non-specific
staining, but it is imperative that you validate the procedure in comparison
to the published method that you are using. This will require you to
Imhojust do it on your tissue and include a no primary control.
If the latter is clear, don't worry about MOM.
If it is not, then we can advise further.
You don't state what Ab/tissue/detection system you are using..
This would help a lot as protocols will vary according to the tissue
Dear Ellen,
You wrote: I'm interested in performing IHC on a mouse ligament.Given
their minuscule size, in lieu of paraffin or cryostat embedding and
sectioning, we would like to embed in plastic and section at 1 micrometer.
Things we are interested in detecting include collagen type 1
Gayle Callis - thanks for the IHC zinc fixative brew - promptly copied
into my permanent files on the subject.
Bob Richmond
___
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Kim,
CD138 is a good marker for plasma cells. Contact Chris van der Loos for a
protocol. He mentioned this at NSH in Denver 2007.
Teri Johnson, HT(ASCP)QIHC
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, MO 64110
Michael: I use 1.5mm nichrome transfer loops for IHC staining of 50um
free floating sections and I move them into clean culture wells for each
solution. I purchase mine from Ted Pella. It does takes some practice
but the loops work quite well with minimal damage to the tissue.
Jean Mitchell,
Rouge, LA 70791
225-763-2564
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mitchell
Jean A
Sent: Friday, July 31, 2009 8:30 AM
To: Michael Patrick; histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE
Apologies if I have not adhered to Histonet protocol: I have changed my Browser
from IE8 to latest Firefox and am not yet familiar!
I agree with Rene: stay with the same wells and give additional, longer washes.
Imho, even experienced users suffer section damage if they change wells.
; this approach
does not require HIER.
2009-07-07
TF
发件人： Colleen Forster
发送时间： 2009-07-07 02:41:59
收件人： Johnson, Teri
抄送： histonet@lists.utsouthwestern.edu
主题： Re: [Histonet] Re: IHC on frozens
I disagree. I do heat retrieval on frozen sections that have been fixed
in a formalin fix
In response to this thread:
snip
Kimberly:
Your question has a two parts answer:
1- it cannot be done because the sections will peel off, but most importantly
2- it is not necessary since HIER was developed to undo the cross linkage
produced by the NBF fixation, and the tissues used for?FS are
I disagree. I do heat retrieval on frozen sections that have been fixed
in a formalin fix. If you use the correct protocol you can get very nice
IHC on these.
Colleen Forster HT(ASCP)QIHC
Anatomic Pathology Research Laboratory
U of MN
Johnson, Teri wrote:
In response to this thread:
We also make our own TBS for our Dako autostainer, same as Jan. I have used
PBS instead of TBS at times, when transfering a bench run where I used PBS /
tween (my standard buffer) to the machine. I wonder if TBS really does give
better results than TBS, when averaged out over all Abs? I do
Hi Meghan,
We had the same problem with Vector VIP - I stain both atherosclerotic
specimens and burns - for some reason the athero samples were fine but the
burns compeltely lost their colour. It turned out, for us, it was the
alcohol in the dehydration step that was stripping the colour. I would
Bond from Leica. It's a great machine and very user friendly. The training
was great and effective. The technical staff helps in so many ways. I've
had my machine for 3 years and would get another one if we increased our
volume.
On 2/2/09 4:16 PM, histonet-requ...@lists.utsouthwestern.edu
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