RE: Daily Digest
Coagulation factor XIIIa stains tissue resident macrophages and activated
macrophages engaged in tissue remodelling. I use 1:400 dulution in PBS with
1% BSA after 30 minute trypsin digestion of FFPE tissue.Calbiochem in La
Jolla Calif sells the rabbit ant human factor IIIa.
Jeff Silverman
Southside Hospital
Bay Shore NY
-----Original Message-----
From: HistoNet Server [mailto:histonet@pathology.swmed.edu]
Sent: Monday, October 01, 2001 11:58 PM
To: HistoNet Server
Subject: Daily Digest
----------------------------------------------------------------------
Date: 1 Oct 2001 03:32:16 -0500
From: Jenny Molde
Subject: Macrophages
Dear Histonetters,
We are currently working with polyurethane synthetic materials and are
having problems with distinguishing between activated and non activated
macrophages. So far we have tried CD68, Ham 56. Is there a specific marker
that would only pick up the above mentioned markers? Any help would be
greatly appreciatted. Many thanks.
Jenny Molde
University of Cape Town
South Africa
----------------------------------------------------------------------
Date: 1 Oct 2001 04:20:23 -0500
From: Phil Bergin
Subject: Re: Macrophages
We use the Dako clone Ber-MAC3 anti human CD163 which seems to be working
quite
well for identifying tissue resident macrophages. Its supposed to be good
for
differentiating between non-activated and activated monocytes.
Philip Bergin
Goteborg Universitet
Goteborg, Sweden
Jenny Molde wrote:
> Dear Histonetters,
> We are currently working with polyurethane synthetic materials and are
> having problems with distinguishing between activated and non activated
> macrophages. So far we have tried CD68, Ham 56. Is there a specific
marker
> that would only pick up the above mentioned markers? Any help would be
> greatly appreciatted. Many thanks.
>
> Jenny Molde
> University of Cape Town
> South Africa
----------------------------------------------------------------------
Date: 1 Oct 2001 04:49:28 -0500
From: Robert Geske
Subject: RE: Macrophages
Phil,
in your experience with the antibody have you seen any correlation between
immunoreactive cells and macrophage cell morphology (ruffeled cytoplasmic
membrane borders, increase vacuolation, nuclear changes) after routine
staining techniques ?
regards,
rob
- -----Original Message-----
From: Phil Bergin
To: Jenny Molde; histonet@pathology.swmed.edu
Sent: 10/1/01 3:42 AM
Subject: Re: Macrophages
We use the Dako clone Ber-MAC3 anti human CD163 which seems to be
working quite
well for identifying tissue resident macrophages. Its supposed to be
good for
differentiating between non-activated and activated monocytes.
Philip Bergin
Goteborg Universitet
Goteborg, Sweden
Jenny Molde wrote:
> Dear Histonetters,
> We are currently working with polyurethane synthetic materials and are
> having problems with distinguishing between activated and non
activated
> macrophages. So far we have tried CD68, Ham 56. Is there a specific
marker
> that would only pick up the above mentioned markers? Any help would
be
> greatly appreciatted. Many thanks.
>
> Jenny Molde
> University of Cape Town
> South Africa
***************************************************************************
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may contain confidential and/or privileged material. If you are not the
intended recipient, please do not read, copy, use or disclose this
communication and notify the sender. Opinions, conclusions and other
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business of my company shall be understood as neither given nor endorsed by
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***************************************************************************
----------------------------------------------------------------------
Date: 1 Oct 2001 05:51:50 -0500
From: "Ryan, Genoula K Ms USAMH"
Subject: IHC
To HistoNetters,
Our new Chief of Pathology would like us to do a few immunos. We are not
set up to do any. Is there a cheap way to do immunos? We don't want to
purchase a Ventanna or anything fancy. Our workload is small and he only
wants the basics (i.e., S100, CK, ER/PR). Any suggestions on how to get
this up and running will be greatly appreciated. Also if there are any good
reference books for IHC. Thanks in advance,
Genie K. Ryan,
Histology Supervisor
HMEDDAC
DSN: 371-2653
49-6221-172653
Genoula.Ryan@hbg.amedd.army.mil
----------------------------------------------------------------------
Date: 1 Oct 2001 06:25:17 -0500
From: Walzer Susan
Subject: RE: Tech Grossing
We have a daily QC checklist that the pathologist signs and one item is
"grossing by non-pathologist". He can check "good", "bad" or write comments.
- -----Original Message-----
From: Jennifer MacDonald [mailto:jmacdona@mtsac.edu]
Sent: Thursday, September 27, 2001 4:29 PM
To: histonet@pathology.swmed.edu
Subject: Tech Grossing
Does anyone use a checklist when a histotech is doing the grossing?
Apparently this is a list that the pathologist can check off when training
or testing the competency of the grossing ability of the histotech. Thank
you in advance.
Jennifer MacDonald
----------------------------------------------------------------------
Date: 1 Oct 2001 07:36:00 -0500
From: Phil Bergin
Subject: Re: Macrophages + 25F9, 27E10 and RM3/1
Rob,
We use the antibody to look at gross numbers of macrophages in bacterial
infected gastric tissue versus non-infected controls. So we haven't really
looked at the individual cell morphology. The 'infected' tissue macrophages
themselves do appear larger and more 'macrophage' like, but this tends to
result in more difficulties in cell counting, we just count relative stained
area of tissue to compensate.
The cells themselves stain variably. Some are beautiful, others have a
patchy
staining with the stain localised to one side of the nucleus.
We have also tried several other antibodies which are supposed to stain at
various stages of activation:
25F9, 27E10 and RM3/1 from Dianova Germany.
They do stain a specific subsets of the CD163 positive cells, but I have a
problem with non-specific and background staining. Don't suppose anyone
else
has any experience with these antibodies?
Phil
Philip Bergin
Goteborg Universitet
Goteborg, Sweden
Robert Geske wrote:
> Phil,
>
> in your experience with the antibody have you seen any correlation between
> immunoreactive cells and macrophage cell morphology (ruffeled cytoplasmic
> membrane borders, increase vacuolation, nuclear changes) after routine
> staining techniques ?
>
> regards,
> rob
>
> -----Original Message-----
> From: Phil Bergin
> To: Jenny Molde; histonet@pathology.swmed.edu
> Sent: 10/1/01 3:42 AM
> Subject: Re: Macrophages
>
> We use the Dako clone Ber-MAC3 anti human CD163 which seems to be
> working quite
> well for identifying tissue resident macrophages. Its supposed to be
> good for
> differentiating between non-activated and activated monocytes.
>
> Philip Bergin
> Goteborg Universitet
> Goteborg, Sweden
>
>
----------------------------------------------------------------------
Date: 1 Oct 2001 08:23:03 -0500
From: Nick_Madary@hgsi.com
Subject: Re: TUNEL Expertise Needed
Here is the number to a self-proclaimed TUNEL expert Frances Swain, she has
been very helpful(although I think we are going to try caspase 3).
501.686.8739 in Little Rock, Ark.
Histo-Scienti
fic Research To: Histonet
Laboratories
Subject: TUNEL Expertise Needed
09/28/01
08:17 PM
Dear Histonetters,
We have a researcher that had sheep myocardial tissue that upon necropsy he
put in 70%ETOH. When he went to do his TUNEL staining, he noted
nonspecific
staining rendering his project useless. Would he be able to 1) cut
sections
from the processed tissue and post-fix the slides in formalin or
paraformaldehyde to make his stain work? 2) If not, could he take the
remaining wet tissue that is currently in 70%ETOH and put it in a fixative
then process as normal and get results with his TUNEL staining? Your help
is greatly appreciated.
Sincerely,
Tom Galati
Histology Laboratory Director
HSRL- A GLP Compliant Contract Laboratory
137 S. Main Street
Woodstock, VA 22664
(540)459-8211
fax: (540)459-8217
tomgalati@hsrl.org
www.hsrl.org
----------------------------------------------------------------------
Date: 1 Oct 2001 10:15:14 -0500
From: "Drew Sally A."
Subject: EBV ISH
My co-worker and I were wondering what the EBer ISH workload volume is out
there for other renal transplant services. We have been pressured to bring
this test onboard, but have also been told by one of the surgeons that the
volume requested would only be 10-12 per year. This seems low, we would
have expected it to be higher since our institution is supposedly the second
largest RTX center in the nation. We're concerned about the cost of
maintaining the test for such low volume-yet we also realize that one of the
issues is the turn-around-time for sending it out.
Sally Ann Drew-MT(ASCP)
IHC/ISH Clin./Res. Laboratory
600 Highland Ave. VAH-DM223
Madison, WI 53792-2472
608-265-6596
sa.drew@hosp.wisc.edu
----------------------------------------------------------------------
Date: 1 Oct 2001 10:31:03 -0500
From: Philip Oshel
Subject: virus alert
Histonetters:
There's another computer virus going around. I suggest going to these
pages and reading about this one, and again *don't* open attachments!
http://www.antivirus.com/vinfo/virusencyclo/default5.asp?VName=TROJ_VOTE.A
http://www.pcmag.com/article/0,2997,s%253D1490%2526a%253D15056,00.asp
- --
Philip Oshel
Supervisor, AMFSC and BBPIC microscopy facilities
Department of Animal Sciences
University of Wisconsin
1675 Observatory Drive
Madison, WI 53706 - 1284
voice: (608) 263-4162
fax: (608) 262-5157 (dept. fax)
----------------------------------------------------------------------
Date: 1 Oct 2001 10:31:19 -0500
From: Dana Settembre
Subject: Re: IHC
On Mon, 1 Oct 2001, Ryan, Genoula K Ms USAMH wrote:
> To HistoNetters,
> Our new Chief of Pathology would like us to do a few immunos. We are not
> set up to do any. Is there a cheap way to do immunos? We don't want to
> purchase a Ventanna or anything fancy. Our workload is small and he only
> wants the basics (i.e., S100, CK, ER/PR). Any suggestions on how to get
> this up and running will be greatly appreciated. Also if there are any
good
> reference books for IHC. Thanks in advance,
> Genie K. Ryan,
> Histology Supervisor
> HMEDDAC
> DSN: 371-2653
> 49-6221-172653
> Genoula.Ryan@hbg.amedd.army.mil
>
>
Genie,
For a cheap way to do immunos I highly recommend that you do them using
the Shandon Sequenza, (which is sort of by hand and not really.) You'll
need their racks and their coverplates. Then I highly recommend that you
use Dako concentrated antibodies for S-100, cytokeratins and ER/PR, their
detection systems and their pretreatment systems, particularly their "Target
Retrieval System"
(TRS). Good Luck!
Dana Settembre
Immunohistochemistry Lab
Pathology Department
University Hospital
Newark, New Jersey
USA
----------------------------------------------------------------------
Date: 1 Oct 2001 10:56:55 -0500
From: Patti Loykasek
Subject: Re: IHC
There are many different avenues you could pursue to start doing immunos.
Dako has a good reference manual that covers IHC staining in general. I am
sure that several different vendors would be happy to help you set up the
basics. You need to decide whether to go with predilutes or concentrated
antibodies based on your expected usage, and your technical knowledge. I
feel that in general, concentrates are best as you can titer for your
specifically processed tissue. Titers are really not that hard to make, and
a good sales rep should help you out with that. Check with your state
histology society for upcoming IHC workshops/seminars. Perhaps even get a
list of laboratories performing IHC and set up a mentoring relationship with
them. If I can answer specific procedural questions, please email me
directly. We perform many types of IHC in our lab, and I would be happy to
try and help. Good luck.
Patti Loykasek
Phenopath Laboratories
Seattle, WA
----------------------------------------------------------------------
Date: 1 Oct 2001 10:57:12 -0500
From: Linda Hall
Subject: Re: TUNEL Expertise Needed
We've been trying other alternatives to the standard TUNEL assay. Trevigen
puts
out a nice TUNEL assay kit which utilizes a
brominated nucleotide (BrdU) which is more efficiently incorporated by
TdT than biotinylated nucleotides at sites of DNA fragmentation. It worked
quite well in my initial studies, and was more specific than the Integren
TUNEL. We've also started doing some work-ups of the Caspase 3 polyclonal
antibody from Cell Signalling.
Histo-Scientific Research Laboratories wrote:
> Dear Histonetters,
>
> We have a researcher that had sheep myocardial tissue that upon necropsy
he
> put in 70%ETOH. When he went to do his TUNEL staining, he noted
nonspecific
> staining rendering his project useless. Would he be able to 1) cut
sections
> from the processed tissue and post-fix the slides in formalin or
> paraformaldehyde to make his stain work? 2) If not, could he take the
> remaining wet tissue that is currently in 70%ETOH and put it in a fixative
> then process as normal and get results with his TUNEL staining? Your
help
> is greatly appreciated.
>
> Sincerely,
>
> Tom Galati
> Histology Laboratory Director
> HSRL- A GLP Compliant Contract Laboratory
> 137 S. Main Street
> Woodstock, VA 22664
> (540)459-8211
> fax: (540)459-8217
> tomgalati@hsrl.org
> www.hsrl.org
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----------------------------------------------------------------------
Date: 1 Oct 2001 11:25:14 -0500
From: Scott Taft
Subject: B5 fixative survey
Hello all,
Would you be kind enough to take this B5 survey?
I appreciate your input in advance.
1. Are you currently using B5 fixative?
2. Has your lab used B5 fixative in the past?
3. If you have discontinued it's use, when and why?
4. What alternative fixative are you using in it's
place?
5. Do you need to use the "de-zenk" procedure for this
substitute?
6. If you use B5; and work in a reference lab, what
percentage of cases require it's use?
Thank you,
Scott Taft HT ASCP
Tucson, AZ
__________________________________________________
Do You Yahoo!?
Listen to your Yahoo! Mail messages from any phone.
http://phone.yahoo.com
----------------------------------------------------------------------
Date: 1 Oct 2001 13:30:56 -0500
From: "Instrumedics, Inc."
Subject: Re: NSH meeting
I want to thank all of you who visited our booth at the NSH meeting. We
enjoyed demonstrating the CryoJane Tape-Transfer and Cryo-Vac-Away systems
for you and to see how they both intrigued you.
I attended a very the fine workshop on Tissue Microarrays. I was pleased to
hear the Paraffin Tape-Transfer system recommended as a way to insure that
an array is captured on a slide. If anyone needs information on how to
maximize the success rate using the PSA system please contact us.
Bernice
schiller@instrumedics.com
----------------------------------------------------------------------
Date: 1 Oct 2001 13:31:11 -0500
From: "Dawson, Glen"
Subject: RE: Forgiving
LaCinda,
There are pompous jerks on any forum such as this. I have decided that
those who would jump all over someone for mis-spelling something or talk
down to any of us for daring to speak without a pHD in hand are the lowest
form of scum. Some of these folks don't except direct emailing (you know
who you are) so direct confrontation can avoid the general histonet post, so
there is little we can do to fight them. Just take heart that there are
more good people than bad in this group and it will get you through.
Glen Dawson BS, HT & IHC (ASCP)
Lead IHC Technologist
Milwaukee, WI
- -----Original Message-----
From: LaCinda Burchell [mailto:ljb@medicine.wisc.edu]
Sent: Thursday, September 27, 2001 12:57 PM
To: tmhpath@amigo.net; Histonet@pathology.swmed.edu
Subject: Re: Forgiving
In the past I have left Histonet twice because I hate the nasty way that
some people treat others on this forum. I keep coming back because I find
the advice on matters pertaining to histology to be very valuable. Does
anybody out there remember the box on a five year old's kindergarten report
card that reads "plays well with others" ? Perhaps more adults should be
asking themselves whether or not that box would be checked in as evaluation
of them. Play nice guys, life is too short to waste all that energy being
mean. Cindy B.
>>> "Michelle D. Moore" 09/25/01 03:20PM >>>
I was on the histonet a year or two ago and I unsubscribed (spelled
correctly) because I found some of my fellow histotechs to be mean and
unforgiving about things. I feel that with all that is going on we should be
more understanding and kinder to those around us. Some people forget a
letter in a word and miss it on spell check. Maybe they are having a bad day
and the snide comments just make things worse. Please lets all try to be
nicer and give people a break for being HUMAN. I enjoy the histonet a great
deal and there is a lot of information for everyone to utilize. I'm sorry if
I offend anyone but I feel we should give each other a break.
----------------------------------------------------------------------
Date: 1 Oct 2001 13:56:53 -0500
From: "Dawson, Glen"
Subject: RE: Florida HT/HTL licensing catch-22
Here here Chuck!!!
Glen
- -----Original Message-----
From: Charles.Embrey [mailto:Charles.Embrey@carle.com]
Sent: Friday, September 28, 2001 5:21 PM
To: 'histonet@pathology.swmed.edu'
Subject: RE: Florida HT/HTL licensing catch-22
Who is everyone kidding. Licensing is the excuse the state uses to take
away more of your hard earned money.
Chuck
- -----Original Message-----
From: Rivera, Maria L. (Tallahassee) [mailto:riveraml@fdhc.state.fl.us]
Sent: Thursday, September 27, 2001 3:38 PM
To: 'Rose Richardson'
Cc: 'HistoNet@pathology.swmed.edu'
Subject: RE: Florida HT/HTL licensing catch-22
What you "see" is a perfect example of not knowing what you are speaking of.
The purpose for licensing in the State of Florida is to make sure that the
quality of care is maintained through regulation and accountability for
those who practice in ANY field of medicine.
In other words, if your livelihood depended on your license you would be
more careful or face ramifications, i.e. lose your license and therefore
your livelihood.
Right now, in most States the only "ramification" around is that a Histotech
would lose their job, then all a Histotech need to do is find another job,
where is the accountability there?
ASCP doesn't take the license away for inept Histotech's. ASCP's concern is
to make sure you are qualified, the "policing" is left to the individual
hospitals. Not so in Florida, the Department of Health takes licensing
seriously as a way to ensure the public of quality of care.
All of us at one time or another have dealt with certain Histotech's that
are the techs from hell. They don't care and other than losing their jobs
they don't pay a price for the inadequacies (headache) they leave the rest
of us, who do care.
If the case was to drive up wages, it certainly did not work in Florida,
this State is probably the lowest paying around.
M
-----Original Message-----
From: Rose Richardson [mailto:rrichar3@iupui.edu]
Sent: Thursday, September 27, 2001 2:43 PM
To: Sharron Ladd;
Subject: Re: Florida HT/HTL licensing catch-22
Licenses are supposed to lead to higher pay and better training, at least
that is the idea that has been tossed around in Indiana. ( with Florida used
as an example!) What I see with this situation you are caught in: the goal
is to limit availability of HT/HTL's to drive the wages up. Big mess for
the ones left on the job, trying to take up the slack when no one can be
hired with license! Who would want to as you say take a year off for a
training program when you have a four year degree and extensive training in
the area? This arrangement is just driving interested people out of this job
market I know that it is said that the training in the school would be
better than on the job. At least where I obtained my degree, I had to have
two semesters of general chem, two of organic chem, two of physics and an
elective from analytical or physical chem!
Rose
- ----- Original Message -----
From: "Sharron Ladd"
To:
Sent: Thursday, September 27, 2001 11:16 AM
Subject: Florida HT/HTL licensing catch-22
Please read the following. Your comments and suggestions would be
greatly appreciated!
1. I want to work as a HT/HTL in the State of Florida.
2. I have a BS degree in biology and 1 year experience working at a
university, in the faculty of medicine working with ANIMAL tissue
(processing, sectioning, staining etc.) which I don't need any special
licenses or ASCP accreditation to do.
3. Because of #2, I qualify to take both the HT and HTL exams offered by
the ASCP BOR and can sign up to do that immediately.
4. Supposing I take and pass the ASCP exam for becoming a HT, I still
need a license to work in Florida.
5. The Florida Department of Health www.doh.state.fl.us states that the
qualifications for Histology technician licensing are as follows:
" For the category of histology, applicants must have 4 hours board
approved HIV/AIDS continuing education and board certification gained by
examination in histology through the Board of Registry of the American
Society of Clinical Pathologists certification program at the
Histotechnician (HT) level with a minimum of a high school diploma or
equivalent AND COMPLETION OF A [Florida DOH] BOARD APPROVED HISTOLOGY
TRAINING PROGRAM (or FOUR years experience plus certification)."
6. If you follow me thus far, here is the catch 22. I must move to
another state to get 4 years of experience (obviously I can't get the
experience here without a license), OR I have to quit my job and sign up
for the training program. There are only 2 places in Florida that offer
the approved training program, Dianon Systems Inc. and Suncoast
Pathology and these training programs are 1 year long.
7. If you are still with me, you might be saying to yourself "oh well,
that's the price you have to pay." However, imagine you are a HT in
another state and you have worked for 3 years as a HT. You want to move
to Florida. You could not get a licence here unless you took the
training program. How ridiculous..who is going to take a training
program for something they have already been thoroughly trained to do?!?
8. SIGNIFICANCE:
Firstly, this could be why I see postings on histonet asking why there
are so many job openings in Florida.
Secondly, this is a major deterrant for people with degrees wanting to
become HT/HTLs. I have already taken 5 years of university in the
biological sciences as well as some Master's degree courses in advanced
molecular techniques. I really like histology but I not interested in
taking a 1 year technician program that I could have taken when I
finished high school.
Thirdly, will this lead to further shortages of HT/HTLs with degrees in
the work force?
----------------------------------------------------------------------
Date: 1 Oct 2001 14:30:38 -0500
From: Abizar Lakdawalla
Subject: Re: NSH meeting
Agree with Bernice, Thom Jensen gave an amazing presentation on tissue
arrays
at the NSH. The amount of support material that he had prepared as well as a
very professional video production showing excatly how to prepare a tissue
array totally blew me away!
This was my first NSH national meeting (got called in to replace someone
else
who could not make it) and I was pleasantly surprised at what a friendly and
informative meeting it was (compared to some of the other national
conferences
that I have attended). It was fun matching faces with names that I knew and
meeting the rest of the vendor community.
Kudos to the organizers,
Abizar
www.innogenex.com
"Instrumedics, Inc." wrote:
> I want to thank all of you who visited our booth at the NSH meeting. We
> enjoyed demonstrating the CryoJane Tape-Transfer and Cryo-Vac-Away systems
> for you and to see how they both intrigued you.
>
> I attended a very the fine workshop on Tissue Microarrays. I was pleased
to
> hear the Paraffin Tape-Transfer system recommended as a way to insure that
> an array is captured on a slide. If anyone needs information on how to
> maximize the success rate using the PSA system please contact us.
>
> Bernice
> schiller@instrumedics.com
----------------------------------------------------------------------
Date: 1 Oct 2001 14:31:00 -0500
From: Gayle Callis
Subject: IEC cryostat, microtome removal
Trying to recyle parts from an old IEC cryostat, pulling microtome out.
Any words of advice (nice ones please!) on how to get the handle off, or
even get this thing dismantled. Sledge hammer is next tool in line here,
unless I hear of something better.
Frustrated and a waste of a good day!
Gayle Callis
MT,HT,HTL(ASCP)
Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)
----------------------------------------------------------------------
Date: 1 Oct 2001 14:31:16 -0500
From: Mark.Lewis@thermoshandon.com
Subject: Thank You !
Hello Histonetters !
It was a pleasure meeting some of you at NSH. It's always good to put a
face to names that are seen on the Histonet.
On behalf of Thermo Shandon, I would like to thank all of you for visiting
our booth and for the pleasure of your company at the cocktail reception
on Monday Evening. It's always good to receive your input (whether good or
bad) concerning Thermo Shandon products.
Thanks again and we look forward to seeing you next year !
Mark Lewis
Product Specialist
Thermo Shandon
1-800-245-6212 ext. 4013
----------------------------------------------------------------------
Date: 1 Oct 2001 16:00:07 -0500
From: Amos Brooks
Subject: Re: IHC
Hi,
DACCA has a new version of their handbook. It is very easy to understand
and fairly short. Theory and troubleshooting are also both covered. I highly
recommend it as a "gettin yer feet wet" book.
If you know anyone in other labs that would be willing to part with
expired reagents, you could get the basics down without spending an arm and
a leg on trial runs. Most commercial IHC kits are very easy to use. Everyone
has a favorite that they swear by. Call a few sales reps and see what works
for you. They should be able to provide you with anything you need in
directions and documentation to start up.
Amos Brooks
- ----- Original Message -----
From: "Ryan, Genoula K Ms USAMH"
To: "'HistoNet Server'"
Sent: Monday, October 01, 2001 6:36 AM
Subject: IHC
> To HistoNetters,
> Our new Chief of Pathology would like us to do a few immunos. We are not
> set up to do any. Is there a cheap way to do immunos? We don't want to
> purchase a Ventanna or anything fancy. Our workload is small and he only
> wants the basics (i.e., S100, CK, ER/PR). Any suggestions on how to get
> this up and running will be greatly appreciated. Also if there are any
good
> reference books for IHC. Thanks in advance,
> Genie K. Ryan,
> Histology Supervisor
> HMEDDAC
> DSN: 371-2653
> 49-6221-172653
> Genoula.Ryan@hbg.amedd.army.mil
>
>
>
----------------------------------------------------------------------
Date: 1 Oct 2001 16:30:17 -0500
From: Gayle Callis
Subject: IEC cryostat is apart
Thanks all,
Found the right tools, cryostat is removed!
Gayle Callis
MT,HT,HTL(ASCP)
Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)
----------------------------------------------------------------------
Date: 1 Oct 2001 16:30:32 -0500
From: Tiffany Sheffield
Subject: bone
I have a really easy question and I feel silly asking it.... What is the
best H&E and Trichrome stain for decalcified paraffin embedded tissue?
Thanks all!
----------------------------------------------------------------------
Date: 1 Oct 2001 17:45:52 -0500
From: Gayle Callis
Subject: Decalcified bone staining
Any H&E works well IF you control the decalcification so that nuclei are
not damaged by acids during decalcification. By control, an endpoint test
(chemical test is easy to do) to determine when calcium is removed and
prevent overexposure of tissue to acid is how one would control
decalcification.
Eosin/Phloxine counterstain gives good tinctoral properties, but make sure
you do not overstain with this, cut back on time. Several companies now
sell this already made up, Richard Allan is one, Newcomer's is another
possibility.
Classic Masson's Trichrome with aniline blue works very well. I used to let
my sections sit overnight in room temperature Bouins rather than use hot
Bouins, sections stayed on slide quite well.
Gayle Callis
MT,HT,HTL(ASCP)
Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)
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Date: 1 Oct 2001 18:30:15 -0500
From: ajennings@unmc.edu
Subject: IHC-cheap (relatively)
I have used the Shandon Sequenza trays/holders for years....use to have the
whole thing and the Cadenza, but it is not necessary if you are just
getting started. Just the coverplates and the trays are needed. I reuse my
coverplates because they are expensive even tho it is not recommended.We
handle them with the greatest of care. The tech time and reagents that you
save more than makes up for the initial cost.I have screened monoclonals
with only 80ul of sample on whole mouse embryo sections. The fact that
gravity works with you not against you (solutions flow by displacement not
capillary action) is a benefit to cleaning up the background. And I am not
sure I understand the theory behind the whys, but I have some antibodies
that I use to use at 1:50 that I can use 1:300 if I use the coverplates.
Please feel free to contact me for more specifics or ask your
Thermo-Shandon rep to give you some info. The system is great for small use
facilities with limited funds. Anita
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Date: 1 Oct 2001 19:15:15 -0500
From: "hpmail.netservers.net"
Subject: Probes for in situ hybridation
Does anyone know where I can get Her 2 neu DNA probes.?
This would be for either fluourescence or non fluorescent type probes. The
later being the most important.
Thanks
----------------------------------------------------------------------
Date: 1 Oct 2001 19:15:30 -0500
From: RSRICHMOND@aol.com
Subject: Re: B5 fixative survey
Scott Taft in Tucson, Arizona asks:
Would you be kind enough to take this B5 survey?
1. Are you currently using B5 fixative? I use it whenever I can in my locum
tenens jobs, and perhaps half of my client - mostly small labs - use it.
2. Has your lab used B5 fixative in the past? I've been using it since it
came into widespread use about 20 years ago.
3. If you have discontinued its use, when and why? - I don't see many labs
that have discontinued it.
4. What alternative fixative are you using in its place? - I've tried
alternatives, with no success. If you can't use B-5 fixative, the best
"alternative" is to insist on overnight fixation of the specimen in neutral
buffered formalin, rather than trying to process it the same day.
5. Do you need to use the "de-zenk" procedure for this substitute? "De-zenk"
- - "dezenkerization" - the use of iodine to remove mercury pigment - is
required for all mercury containing fixatives.
6. If you use B5; and work in a reference lab, what percentage of cases
require its use? - I don't work in a reference lab, at least not often. A
very low percentage of cases. More if you fix bone marrow biopsies in it.
The
important thing is to use as little of it as possible - no more than 10 mL
per case.
Bob Richmond
Samurai Pathologist
Knoxville TN
----------------------------------------------------------------------
Date: 1 Oct 2001 19:41:45 -0500
From: Cathy Mayton
Subject: NSH Convention
Fellow Histonetters,
I wanted to thank all the folks at NSH, and all the volunteers who helped
put on a great convention in Charlotte!! In spite of the recent national
tragedy, the Convention seemed to go on without a hitch. It did seem
smaller than previous Conventions but the AV folks and all who helped with
the Convention did a super job.
I also wanted to thank the vendors who came and exhibited all the new
equipment and consumables. Thank you for all for continuing to support NSH.
I missed many of my colleagues who did not attend for whatever reason, but
hopefully next year will be better for everyone and the Long Beach
Convention will be a whopper!!
Take care,
Cathy
*********************************
GLP Compliant Laboratory
Cathy A. Mayton
Project Director
Wasatch Histo Consultants, Inc.
www.wasatchhisto.com
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Fellow Histonetters,
I wanted to thank all the folks at NSH, and all the
volunteers who helped put on a great convention in Charlotte!!
In spite of the recent national tragedy, the Convention seemed to go on
without a hitch. It did seem smaller than previous Conventions but
the AV folks and all who helped with the Convention did a super
job.
I also wanted to thank the vendors who came and exhibited all the new
equipment and consumables. Thank you for all for continuing to
support NSH.
I missed many of my colleagues who did not attend for whatever reason,
but hopefully next year will be better for everyone and the Long Beach
Convention will be a whopper!!
Take care,
Cathy
*********************************
GLP Compliant Laboratory
Cathy A. Mayton
Project Director
Wasatch Histo Consultants, Inc.
www.wasatchhisto.com
- --Boundary_(ID_q4ICEc0otHXxjImFb5fPGg)--
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Date: 1 Oct 2001 20:30:36 -0500
From: Richard Cartun
Subject: Re: Probes for in situ hybridation
I would try Zymed Laboratories (www.zymed.com) or (800) 874-4494.
R. Cartun
>>> "hpmail.netservers.net" 10/01/01 07:56PM >>>
Does anyone know where I can get Her 2 neu DNA probes.?
This would be for either fluourescence or non fluorescent type probes. The
later being the most important.
Thanks
----------------------------------------------------------------------
Date: 1 Oct 2001 20:44:26 -0500
From: Richard Cartun
Subject: IHC-Reply
Hi Genie:
I think before you embark on setting-up IHC in your laboratory you need to
answer a few questions regarding your IHC workload and your (and your
pathologist's) time commitment to perform quality control/assurance
activities
relating to IHC staining. I see too many problems (including mis-diagnoses)
from labs that are not performing these procedures on a daily basis. Also,
keep in mind that therapeutic decisions are being established on the results
of IHC staining (especially ER/PR) and you better be able to show that you
are
producing a quality product that is reproducible time after time. Just a
few
words of caution............
R. Cartun
----------------------------------------------------------------------
Date: 1 Oct 2001 23:00:48 -0500
From: Debbie Pepperal
Subject: Re: In Situ for EBV
Hi Dana
We use the EBV mRNA from Novocastra. Works great.
Zenobia Haffajee
At 11:17 AM 28/09/2001 -0400, you wrote:
>What are some commonly used kits out there for In Situ for EBV on FFPE
>human tissue?
>What are the vendors? Help me?
>
>Dana Settembre
>Immunohistochemistry Lab
>Pathology Department
>University Hospital
>Newark, New Jersey
>USA
>
>
>
----------------------------------------------------------------------
Date: 1 Oct 2001 23:13:45 -0500
From: Debbie Pepperal
Subject: Re: Probes for in situ hybridation
Hi
Ventana have FISH probes. to HER2neu..however it is an automated system and
Vysis have FISH probes too.
Zenobia Haffajee
At 04:56 PM 01/10/2001 -0700, you wrote:
>Does anyone know where I can get Her 2 neu DNA probes.?
>
>This would be for either fluourescence or non fluorescent type probes. The
>later being the most important.
>
>Thanks
>
>
>
----------------------------------------------------------------------
Date: 1 Oct 2001 23:27:42 -0500
From: Debbie Pepperal
Subject: MIB-1 replies
Hi Histonetters
thank you to all for the quick response to my MIB-1 query.
Zenobia Haffajee
Here are the messages received yesterday!
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