Does anyone out there know anything about doing IHC on skin? Any published
articles?

Melissa Brennan
Silver Spring, MD

-----Original Message-----
From: HistoNet Server [mailto:HistoNet@Pathology.swmed.edu]
Sent: Saturday, March 27, 1999 11:00 PM
To: HistoNet Server
Subject: Daily Digest



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

Date: 27 Mar 1999 00:11:07 -0600
From: "J. A. Kiernan" <jkiernan@julian.uwo.ca>
Subject: Silver stain removal (& other thoughts. Was: Thanks)

On Fri, 26 Mar 1999, Inka Tertinegg wrote:

> Does anyone know how to get silver stain out of
> a white benchtop?

  Inka:  It depends how tough the bench top is.

    1. Nitric acid will remove silver (but not gold).
       Dilute the conc. acid about 50/50 with water.
       OK on epoxy paint. I don't know about formica,
       but would guess probably safe at this dilution.

    2. (From back in the old days) A paste of mercuric
       chloride and water rubbed into the black silver
       stain. More gentle on the skin than nitric acid,
       I've been told. Never really an acceptable method.

    3. Farmer's reducer, as used in photography; but for
       cleaning purposes it needs to be more concentrated.
       Crunch up about one cubic centimetre each of
       potassium ferricyanide and sodium thiosulphate in
       15-20 ml of water. Apply generously to the blackened
       bits of bench. It will remove the silver but not
       quickly. Allow 30-60 minutes and don't let it dry
       out. The slowness is a penalty for (a) spilling
       your expensive silver solution, and (b) using fairly
       harmless chemicals.
         Be sure to use ferrIcyanide and NOT ferrOcyanide,
       which will not work. Despite the -cyanide in their
       names, these are not considered dangerous poisons.

         (Cyanides, well known for their speed, are much more
          potent and also easily available to murderers in
          detective stories. When I was a lad, one of the
          other students in the lab needed to order some
          KCN for histochemical purposes. He needed about
          50 mg but the stuff was cheap and the smallest
          bottle held 500 g. We looked it up in Polson's
          Toxicology and found that this would be enough to
          kill 1500 people, if they could all be
          persuaded to swallow their 333 mg helpings. The
          smell must surely make cyanide a rather poor
          weapon for the enterprising poisoner: "bitter 
          almonds" to the novelist, but "sickly sweet" to 
          anyone working with amounts well below 10% of what's 
          supposed to be lethal.)

    4. None of these methods will remove dark stains due to
       metallic gold, from spilt "gold chloride."  Cyanide
       and oxygen would, and so would aqua regia, but these
       are not for sploshing on the bench's top. To hide
       carelessness with gold, you'll need a pot of paint.

           John A. Kiernan,
           Department of Anatomy & Cell Biology,
           The University of Western Ontario,
           LONDON,  Canada  N6A 5C1




     



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

Date: 27 Mar 1999 05:30:29 -0600
From: marsdijk@wxs.nl
Subject: usersr of PAPnet

Are there any users of the PAPnet, and what are they going to do now NSI
doesn't do to well ?

- --
met vriendelijke groet,



 John



*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*

|  John van Marsdijk            |

*  email: marsdijk@wxs.nl       *

|  Veenslag 31                  |

*  3905 SJ  Veenendaal          *

|  The Netherlands  |

*                               *

|  tel:   (+31) 06-53 44 2176   |

*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*




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

Date: 27 Mar 1999 07:45:39 -0600
From: Valleygal@aol.com
Subject: ASH Workshop

The Arizona Society for Histotechnology is proud to announce the second in a
series of workshops to be given on April 17, 1999 at Scottsdale Healthcare -
Osborn Campus. The topic of this workshop is Her2neu by IHC, Prognostic and
Therapeutic Values (3.0 contact hours) and is being sponsored by IMPATH and
Dako Corp. The speaker is Daisey Jimenez Joseph, B.A., HTL (ASCP).
The workshop will be held in the conference room at 7300 E. 4th Street and
will begin at 10AM. 
A lunch will be served courtesy of IMPATH and Dako.
There is no charge to ASH members and a $5.00 charge for non-members. 
RSVP by April 13, 1999 by contacting Carole Reeder at 602-481-4000, ext.
2199.
Space is limited for this wet workshop and lecture so make your reservations
early.
See you there!

Andi Grantham


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

Date: 27 Mar 1999 08:30:59 -0600
From: "Sandy Julsing" <sandy.julsing@unitedhealth.org>
Subject: processing GI biopsies -Reply

Kathleen,
   How do you warm your forceps for embedding?  Even if GI biopsies are
fixed
on time, processed gently and oriented carefully, if a flame or incinerator
(400 degrees!) are used to embedd and you go directly from these over heated
sources to your tiny unsuspecting gastric biopsy, you can sizzle you tissue
instantly.
    Just a thought!

>>> Kathleen Hollenbeck <lastandfirst@webtv.net> 03/25 10:41 pm >>>
We process our GI biopsies on a short cycle using a VIP tissue
processor.  Our results are not consistent.  Somedays we experience a
lot of microchatter and other days we do not.  We receive specimens from
out-patient locations in addition to our in-house surgical specimens.

Any helpful hints will be greatly appreciated.  Thanks in advance.

Kathleen Hollenbeck
Histology Supervisor
Sunrise Hospital
Las Vegas, NV





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

Date: 27 Mar 1999 09:15:48 -0600
From: "Gary W. Gill" <garywgill@email.msn.com>
Subject: RE: usersr of PAPnet

FYI, Neuromedical Systems voluntarily filed on 03/26 for bankruptcy
protection in a Wilmington, Delaware court and "therewith let go the
majority of its US workforce."  NeoPath, maker of AutoCyte, will acquire
NSI's "patent estate" and 1.4 million shares of common stock for $4 million,
pending approval.  The recently published "Duke evidence report" and JAMA
article (too little bang for the buck) probably accelerated the inevitable
demise.

Perhaps some large lab will acquire numerous deeply discounted PapNet
devices and offer a regional Pap smear screening service.  However, the
latter is what got NSI into trouble in the first place.  Or perhaps, a few
university researchers will use PapNet to conduct cytological studies.  But
where will they get the spare parts?  It's not a happy situation.

I am not a PapNet user, though I took their week long training course in
1997.  The technology is impressive, but NSI's early marketing strategy and
tactics were not well received by potential end users.  NSI persisted in
this strategy for too long to recover.  Like all automated systems, PapNet
is not perfect; it still misses abnormal cells.  It is also very expensive.
PapNet driven screening microscopy systems are very sophisticated
electronically and require daily QC/QA measures -- which is to be expected.

I wish all NSI employees well.  The few I met were competent, conscientious,
personable individuals.  Having experienced sudden and unexpected
outplacement twice in the past 10 years, I appreciate firsthand the turmoil
created in one's personal and professional life.  Becoming re-employed is
especially difficult when one's job skills are very specialized and matching
job opportunities are few and far between.

Gary Gill

> -----Original Message-----
> From: John van Marsdijk [mailto:marsdijk@wxs.nl]
> Sent: March 27, 1999 3:11 AM
> To: Histonet listserver
> Subject: usersr of PAPnet
>
>
> Are there any users of the PAPnet, and what are they going to do now NSI
> doesn't do to well ?
>
> --
> met vriendelijke groet,
>
>  John




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

Date: 27 Mar 1999 10:00:22 -0600
From: marianne brown <mbrown_biocaremedical@yahoo.com>
Subject: Re: CD-19



- --- "Nocito, Joseph" <joseph_nocito@srhc.iwhs.org> wrote:
> Hello Histo land,
> does any one know of a CD-19 antibody that works in
> paraffin.  My
> pathologists want another B-cell marker other than
> CD-20.  As always, thanks
> for your help.
> 
> Joe Nocito, B.S., HT(ASCP)QIHC
> Histology Supervisor
> Christus Santa Rosa Hospitals
> San Antonio, Texas 
Biocare Medical 1-800-799-9499 has a coctail B cell marker L26/CD20
Marianne
> 
> 
> 

===
Marianne Brown
Consultant Biocare Medical 


_________________________________________________________
Do You Yahoo!?
Get your free @yahoo.com address at http://mail.yahoo.com



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

Date: 27 Mar 1999 10:00:51 -0600
From: "D. Hammer" <hammerd@u.washington.edu>
Subject: Histonet Archives 

Marvin,

I just spent a great time on the site you inform us about below.
Fantastic job!  It was so easy to use and will be so helpful to us when
gathering info on specific topics.  I have noticed several messages on
histonet begin with ...."I know this topic has been discussed before, but
......."   Now we can just go to your site and extract the info you have
stored.

Thanks again for a terrific addition the the Histonet Service.  What a
world we live in :)

Don
PS  A tip for Histonetters who access the page by copying the
address....becareful not to include the period after the address, took me
a couple of try's before figuring out why it wouldn't open.
                                              


- ---------- Forwarded message ----------
Date: Mon, 08 Mar 1999 19:55:55 -0500
From: Marvin Hanna <mhanna@magicnet.net>
To: histonet@Pathology.swmed.edu
Subject: Histonet Archives


Hi Histonetters,

We have been running a search engine for pathologists and laboratory
professionals for the past 6 months at http://www.pathsearch.com. A few of
my friends and co-subscribers on Histonet  have asked if we could archive
Histonet, so...

There is now an archive of Histonet messages since October, 1998 on the web
server at http://www.pathsearch.com/histonet.html. They are available as
either an archive by dates or searchable by the search engine. The web
server updates the archive hourly.

We have also searched and indexed a number of other histology resources on
the Internet and provided a specialized search engine for them and
Histonet. You can search sites either collectively or individually at
http://www.pathsearch.com/histosearch.html. Histosearch is a subset of over
10, 000 pages that is included in the over 50,000 pages searched and
indexed on Pathsearch.

We have also set up a 24 hour a day chat server, the ability to publish
home pages or technical papers on the Internet in a couple of minutes, and
intelligent agent technology for both Pathsearch and Histosearch.

The Histonet archives, Histosearch, and Pathsearch are free to
histologists, pathologists, and laboratory professionals on the Internet.
We do plan on offering sponsorships on Histosearch and Pathsearch to help
cover the cost of colocating the server on a T-3 line, hardware, software
and time.

Linda and Herb have done a great service for the field of histology in
setting up and running Histonet. I hope archiving this information will
benefit the participants of Histonet.

Marvin Hanna
webmaster@pathsearch.com









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

Date: 27 Mar 1999 10:01:15 -0600
From: "Nocito, Joseph" <joseph_nocito@srhc.iwhs.org>
Subject: CD-19

Thanks to those who responded to my CD-19 question.  I told my pathologists
that there wasn't a CD-19 that works in paraffin.  The way to go seems to be
CD-79a.  I'll give that a try and see if the pathologists will be satisfied.
Thanks again

Joe Nocito, B.S., HT(ASCP)QIHC
Histology Supervisor
Christus Santa Rosa Hospitals
San Antonio, Texas 



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

Date: 27 Mar 1999 10:30:33 -0600
From: "D. Hammer" <hammerd@u.washington.edu>
Subject: Correction on Medicare Billing


Histonetters,

A couple of months ago, in response to a discussion on Medicare Billing, I
posted our way of handling the 72 hour rule when receiving specimens from
outside clients.  (I stated to comply with the Rules we were going to have
to bill the institution for the Technical Component on these as the
payment is included in the DRG paid to the orriginating Hospital.)

More recent information (an overlooked Rule) has cleared the air.  When
the DRG's were designed, for some reason Anatomical Pathology fees were
not included. Therefore Medicare may be billed for the Technical Component
by the institution performing the work.  The Professional Fee has always
been understood to be billed to Medicare.

This really lifts a burden off the billing process for both the submitting
and performing institutions.

I sure hope the APG's (APC's) will be set up in the same way for Out
Patients or we may have to bill differently when they are in place. :(

Don  
PS  I am at home and do not have the Reference # but will send it out on
Monday.                                             
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Don Hammer, Administrative Director            UNIVERSITY OF WASHINGTON 
Hospital Pathology, Box 356100                     MEDICAL CENTER
1995 NE Pacific St.                                
Seattle Washington, 98195                  ~Where Knowledge Comes To Life~ 
(206) 548-6401 Fax: (206) 548-4928         
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~







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

Date: 27 Mar 1999 11:45:41 -0600
From: "Douglas A. Porter" <porter.doug@acd.net>
Subject: Histonet Archive

Marvin Hanna wrote:

Hi Histonetters,
We have been running a search engine for pathologists and laboratory
professionals for the past 6 months at http://www.pathsearch.com. A few of
my friends and co-subscribers on Histonet  have asked if we could archive
Histonet, so...
There is now an archive of Histonet messages since October, 1998 on the web
server at http://www.pathsearch.com/histonet.html. They are available as
either an archive by dates or searchable by the search engine. The web
server updates the archive hourly.
We have also searched and indexed a number of other histology resources on
the Internet and provided a specialized search engine for them and Histonet.
You can search sites either collectively or individually at
http://www.pathsearch.com/histosearch.html. Histosearch is a subset of over
10, 000 pages that is included in the over 50,000 pages searched and indexed
on Pathsearch.
We have also set up a 24 hour a day chat server, the ability to publish home
pages or technical papers on the Internet in a couple of minutes, and
intelligent agent technology for both Pathsearch and Histosearch.
The Histonet archives, Histosearch, and Pathsearch are free to histologists,
pathologists, and laboratory professionals on the Internet.  We do plan on
offering sponsorships on Histosearch and Pathsearch to help cover the cost
of colocating the server on a T-3 line, hardware, software and time.
Linda and Herb have done a great service for the field of histology in
setting up and running Histonet. I hope archiving this information will
benefit the participants of Histonet.
Marvin Hanna
webmaster@pathsearch.com


Marvin,

I have been archiving selected Histonet emails since November of 1997.  I
have 458 to date!  If you would like to somehow add those to yours I would
be happy to assist in any way I can.  Thanks and good luck!

Doug
porter.doug@acd.net





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

Date: 27 Mar 1999 11:55:42 -0600
From: "Alexander Nader" <Alexander.Nader@univie.ac.at>
Subject: Re: CD-19

On Sat, 27 Mar 1999 09:51:01 -0600, Nocito, Joseph wrote:

>Thanks to those who responded to my CD-19 question.  I told my pathologists
>that there wasn't a CD-19 that works in paraffin.  The way to go seems to
be
>CD-79a.  I'll give that a try and see if the pathologists will be
satisfied.
>Thanks again

Be carefull with CD79a: I've seen a couple of T-cell lymphomas (mostly
peripheral large 
cell variant) staining very nicely with CD79a.


			Alexander Nader
			Alexander.Nader@univie.ac.at
			HTTP://members.teleweb.at/anader 



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

Date: 27 Mar 1999 11:56:07 -0600
From: "Alexander Nader" <Alexander.Nader@univie.ac.at>
Subject: Re: AUA1

On Fri, 26 Mar 1999 11:37:40 -0800, Technical Services wrote:

>Hi Dave,
>This is listed as a neoplastic epithelial marker, and I show that
Bionostics
>has it, their number is 508-263-3856. I also have a listing for a
>Bionostics, Inc and in case this is the company you need, their number is
>416-789-3936.
>

Does anybody know if this antibody is also sold in Europe?

			Alexander Nader
			Alexander.Nader@univie.ac.at
			HTTP://members.teleweb.at/anader 



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

Date: 27 Mar 1999 13:15:21 -0600
From: "Hewlett Bryan  (CMH)" <HEWLETT@EXCHANGE1.CMH.ON.CA>
Subject: RE: CD-19

Alexander,

Is that anecdotal or a published observation?
We have only seen anomalous reactions with CD79a on a couple of
occasions, both times with epithelia!
But then again, it's possible to stain anything with anything if you
really try hard!!

Bryan

>----------
>From: 	Alexander Nader[SMTP:Alexander.Nader@univie.ac.at]
>Sent: 	March 27, 1999 12:45 PM
>To: 	Histonet; Nocito, Joseph
>Subject: 	Re: CD-19
>
>On Sat, 27 Mar 1999 09:51:01 -0600, Nocito, Joseph wrote:
>
>>Thanks to those who responded to my CD-19 question.  I told my
pathologists
>>that there wasn't a CD-19 that works in paraffin.  The way to go seems to
be
>>CD-79a.  I'll give that a try and see if the pathologists will be
satisfied.
>>Thanks again
>
>Be carefull with CD79a: I've seen a couple of T-cell lymphomas (mostly
>peripheral large 
>cell variant) staining very nicely with CD79a.
>
>
>			Alexander Nader
>			Alexander.Nader@univie.ac.at
>			HTTP://members.teleweb.at/anader 
>
>
>


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

Date: 27 Mar 1999 22:45:42 -0600
From: "Masayuki Miyagishima, MD" <mmiyagis+@pitt.edu>
Subject: mice frozen tissue IHC protocol.

This is a very fundamental question.
What is the best or standard IHC protocol for FROZEN MICE heart and
vessel?
They are 4% paraformaldehyde perfusion fixed and sit for 2 hours in ice
cold 4 % paraformaldehyde and then treated with 30 % sucrose for
overnight, then embedded in OCT and frozen in Isopentan.  The tissue is
limited and small, but we need to do many IHCs.

1) When we cut the frozen tissue samples, we cut about 50 sections at
once and put them into -70 fridge for the future IHCs.  Do you think I
should refix them and air-dried before putting them into -70?

2) After taking out from -70 fridge, we treat the sections with 10% Goat
serum, then treat with rabbit primary antibody for 2 hours at RT, then
with goat anti-rabbit secondary antibody for 2 hours at RT.  Do you have
any modifications for mice tissue?

Thank you in advance.

- -- 
Masayuki Miyagishima, MD
University of Pittsburgh, 
Department of Surgery
412-647-2345, and ask the hospital operator to page 3228, please.


Here are the messages received yesterday!