RE: histonet button

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From:"Culberson, Cathy" <>
To:'HistoNet Server' <>
Date:Fri, 22 Oct 1999 07:18:50 -0400
Content-Type:text/plain; charset="iso-8859-1"

Please send a button this way as we were not allowed to travel.
Thanks so  much.
Cathy Culberson, MS
Supervisor, General Surgery Research

	-----Original Message-----
	From:	HistoNet Server []
	Sent:	Friday, October 22, 1999 1:01 AM
	To:	HistoNet Server
	Subject:	Daily Digest


	Date: 21 Oct 1999 00:09:41 -0500
	From: Kathy Gorham <>
	Subject: staffing survey

	Hi, I'm working on  my budget for next year and would like to show a
	justification for  an increase in staffing.  If you could take the
time to
	fill out this survey I would greatly appreciate it.

	Processing                     #per year

	Special Stains

	Staffing	            # hrs per day


	Do you assist/provide the following Services
	Frozen Sections
	Billing data entry
	Slide/Block Filing

	Thanks, Kathy Gorham, HT
		  Grande Ronde Hospital
		   LaGrande, Or. 97850


	Date: 21 Oct 1999 02:30:39 -0500
	From: "Jim Manavis" <>
	Subject: Progesterone receptor

	Can any one provide me with information on what Progesterone
receptor clones
	they are using. Which is the clone of choice and why?

	We currently use an Abbott rat anti human (clone KD68) on our FFPE
	material, which we enhance. Apart from this we get a very good

	I am currently assessing the Dako mouse monoclonal PgR (clone PgR
636). The
	staining pattern looks very similar, but with a much more florid
	and at a much better dilution.

	If PgR 636 is the marker of choice could someone supply me with the
	appropriate references/verification.



	or via Histonet


	Date: 21 Oct 1999 02:31:03 -0500
	From: "Sarah A. Jones" <>
	Subject: Histonet Buttons

	Once again I was chained to my friendly exhibitor's booth, and was
unable to
	even get upstairs to get a Histonet Button!  In fact I didn't even
	where to find them until Monday, when I finally discovered someone
who knew
	where to find them just before I had to leave for the airport!!

	So....if you would kindly consider sending one to me I would really
	appreciate it!

	Sarah A. Jones
	540 N. Santa Cruz Ave.
	PMB #297
	Los Gatos, CA 95030


	Date: 21 Oct 1999 08:45:21 -0500
	From: Cynthia Favara <>
	Subject: RE: antibody to EBV antigen LMP-2

		The only antibody I know of with those initials is Lysosomal
	Protien and they are numbered, don't know of any connection with EBV
	Cynthia Favara
	903 South 4th Street
	Hamilton, MT 59840
	PH: 406-363-9317
	FAX: 406-363-9286


	Date: 21 Oct 1999 08:45:46 -0500
	From: "Tony Henwood" <>
	Subject: Re: IF

	Dear Sally,

	>  Has anyone ever performed IF on formalin fixed paraffin embedded
tissue? If
	> yes. Is there an enzyme digesting agent that is better to use for
IF? Also I
	> think I read somewhere that 10% Neutral Buffered Formalin
	> Has anyone heard this?

	You can do IF on formalin fixed tissue just like a regular IPX (it
	only the label that is different). Use enzyme or antigen retrieval
	usual. IF on formalin fixed skin and renal (like IPX) is difficult 
	but not impossible.

	Collagen will fluoresce after formalin fixation and of course
	will as well (basis of the formalin induced fluorescence test). The 
	appropriate use of filters will decrease this problem.

	Regards, Tony

	Tony Henwood
	Senior Scientist
	Anatomical Pathology
	Royal Prince Alfred Hospital


	Date: 21 Oct 1999 10:00:48 -0500
	From: "Jennings, M. Anita" <>
	Subject: FW: FW: NSH Diary, Final

	Last one for this year. Thanks a bunch Andi. Bring back all of your
	acquired knowledge and share with the rest of us. anita

	OK Anita, one more entry for NSH 1999:
	It was a dark and rainy nite...well it was! Cold too.
	Workshops are over, and all the signs of NSH gone from the
	It always seems sad to have witnessed all the excitement of a few
days ago
	gone for another year. Now everyone is looking forward to going home
	such a great week of educational opportunities taking with them all
	things they learned from their colleagues.
	The banquet was well attended and I can't remember the all award
	but I couldn't help but feel as the accomplishments of each
recipient was
	read off that these people are extremely deserving of their
recognition by
	NSH. One award I do remember was a special award to Linda and Herb
	us Histonet! Congratulations Linda and Herb!!!
	I don't know who won time contest tonight. =;-)
	Tomorrow is the HOD Meeting - the society at work.
	This has been a memorable week, but Anita, I hope that you make it
	Wisconsin next year so you can continue on the NSH Diary tradition.
	sending a histonet button home for you.
	Over and out.



	Date: 21 Oct 1999 10:30:52 -0500
	From: "Sebree Linda A." <>
	Subject: RE: antibody to EBV antigen LMP-2

	We use Cell Marque's LMP (latent membrame protein) cocktail which
	of 4 clones:  CS1, CS2, CS3 and CS4.  I wonder if by LMP-2, you are
	referring to the CS2 clone?  I don't know that I've seen these
clones as
	separate antibodies but they probably are marketed as such
somewhere.  Maybe
	one of the vendors out there can tell you.

	Linda A. Sebree, HT
	University of Wisconsin Hospital & Clinics
	Immunohistochemistry/In Situ Hybridization Laboratory
	600 Highland Avenue
	Madison, WI  53792-2472

	FAX: (608)263-1568

	> -----Original Message-----
	> From:	Sarka Lhotak []
	> Sent:	Wednesday, October 20, 1999 7:33 PM
	> To:
	> Subject:	antibody to EBV antigen LMP-2
	> Hello all,
	>         Has anybody come across an antibody to the EBV antigen
LMP-2? Any
	> lead would be greatly appreciated. Thanks,
	> Sarka Lhotak
	> Hamilton Regional Cancer Centre
	> McMaster University
	> Hamilton, Ontario


	Date: 21 Oct 1999 11:45:34 -0500
	From: alexis <>
	Subject: Hypoxia in mouse tissue

	Hi, all!
	I am looking for a good antibody to visualise hypoxic cells in
	embedded  mouse tumors. So far I've only found HIF-1alpha from
	but it hasn't been tested for immunos.  I also found a reference to
	(2- nitroimidazole theophyllin) developed at Brunel University, UK.
but no
	mention to vendors or how to obtain it.
	I would really appreciate any help!

	Center for Transgene Research
	and the
	Department of Chemistry and Biochemistry
	University of Notre Dame
	Notre Dame, IN, USA
	219-631-3539 (phone)
	219-631-4048 (fax)


	Date: 21 Oct 1999 11:45:58 -0500
	From: Jo-Ann Bader <>

	Hi all ,

	I am looking for a stain specifically for mitocondria on paraffin
	Is there such a thing?

	Thanks in advance


	Date: 21 Oct 1999 12:46:00 -0500
	From: "J. A. Kiernan" <>

	On Thu, 21 Oct 1999, Jo-Ann Bader wrote:

	> I am looking for a stain specifically for mitocondria on paraffin
	> Is there such a thing?

	  Yes indeed. Mitochondria were discovered by staining them
	  in paraffin sections, and methods are given even in modern
	  techniques books. I haven't tried all the methods, but have
	  seen mitochondria using Baker's acid-haematein (a histochemical
	  method for choline-containing phospholipids) and with Bensley's
	  copper-haematoxylin, a traditional method. More ordinary dyes 
	  will also stain mitochondria, but not selectively enough to
	  see them properly. The most colourful selective method is
	  said to be Altmann's aniline-acid fuchsine-picric acid, but
	  I've never tried that one.

	  Mitochondria can be stained only if they have been properly
	  fixed before embedding. A mitochondrion is mostly membrane, so
	  it's necessary to make the phospholipids insoluble before
	  dehydration, clearing and embedding. This can be done by a
	  treatment with osmium tetroxide or potassium dichromate (or
	  both) after primary fixation in neutral buffered formaldehyde,
	  with or without added glutaraldehyde. Helly's fluid also
	  preserves mitochondria. The traditional fixatives for cytoplasm
	  and organelles are unbuffered mixtures of OsO4 with K2Cr2O7,
	  used on very tiny pieces of tissue. The sections need to be
	  thin - 4 mu or less.

	  The selective stains for mitochondria also colour red blood
	  cells and myelin sheaths, but these do not cause confusion.

	  It's good to hear that there's still a demand for traditional
	  staining methods for mitochondria.

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


	Date: 21 Oct 1999 13:45:59 -0500
	From: (Marija Vaicaitis)
	Subject: rapid AE1/AE3

	We routinely do an AE1/AE3 on all sentinel nodes. But now the
question came
	up-if there is a possibility of doing a rapid
	AE1/AE3 on frozen section slides.   I would appreciate any

	Marija Vaicaitis
	Pathology Lab
	Little Co. of Mary Hospital
	Evergreen Park, Il


	Date: 21 Oct 1999 14:00:37 -0500
	From: "Kellar, Eric" <kellarec@MSX.UPMC.EDU>

	The mitochondrion was identified at the end of the 19th century by
	Benda in 1903 coined the term mitochondrion, from mito, thread, and
	khondrion, granule, because these organelles appeared as
	granules under the light microscope. Only a few years later, most of
	relevant functions of mitochondria, such as energy production, cell
	respiration and inheritance, were suggested by Michaelis, Regaud and
	Kingsbury. Even the biological origin of mitochondria as a symbiotic
	between a prokaryote and a primitive eukaryote cell was hypothesized
	Portier decades before it could be more firmly established.

	The next development in mitochondrial staining after Altmann in
1890, was
	the technique developed by Kull in 1914 using the highly explosive
	aurantia.  Kull also substituted the fixative of Champy 1913 for the
	fixative specified by Altmann in 1890. On the basis that Champy
	this fixative for another purpose, the technique has been commonly
known as
	the Champy-Kull technique. (any technique in which acid fuchsin is
used as
	the primary stain). The two techniques of Champy-Kull and Parat  (in
	sections are "pre-dichromated") were brought together in 1928 by

	The more modern and commonly used techniques are given by Cain (AFIP
	and another by Takaya, both of which treat the sections in Regaud's
	prior to staining. 

	I have tried both of these methods (preferring that of Cain) and
would be
	glad to fax you a copy of either.

	Eric C. Kellar
	University of Pittsburgh Medical Center

		From: 	Jo-Ann Bader[]
		Sent: 	Thursday, October 21, 1999 12:27 PM

		Hi all ,

		I am looking for a stain specifically for mitocondria on
		Is there such a thing?

		Thanks in advance


	Date: 21 Oct 1999 14:23:41 -0500
	Subject: CD31 and CD10 

	Could some tell me a good CD31 and CD10 antibody  for formalin-fixed
	paraffin embedded tissues.  What procedures and pretreatments are



	Date: 21 Oct 1999 14:25:18 -0500
	From: Atoska Gentry <>
	Subject: cryostat

	Histonetters, especially those of you who replied to my previous
	you who recommend the Hacker/Bright cryostat which models do you
have in
	use and is your lab setting one of clinical or research?  After a
	research I've learned that Hacker manufactures a wide range or
	and I'm trying to decide which one will best suit our needs. Also
I'd like
	to know your likes and dislikes in reference to your experience with
	you can email me personally if you do not wish to do so on the
	Thanks, Atoska

	Atoska S. Gentry B.S., HT(ASCP)
	Research Assistant
	Scott-Rtichey Research Center
	College of Veterinary Medicine
	Auburn University, AL  36849
	PH (334) 844-5579
	FAX (334) 844-5850


	Date: 21 Oct 1999 15:21:51 -0500
	From: "pathanjaly hariharan" <>
	Subject: CD31 and CD10

	Hello Dave,
	InnoGenex carries an anti-CD31(Clone 9G11) which is
	validated for paraffin sections, as well as frozen.
	For paraffin embedded tissues: Antigen Retrieval
	Solution AR10(pH10) is recommended. 
	Primary incubation = 2 hours at 37 degrees C.

	The data sheet can be accessed at:
	InnoGenex: (877)IGX INFO

	If you would like more information, please give me a
	Best Regards,
	Matthew Ogdie

	Could some tell me a good CD31 and CD10 antibody  for
	paraffin embedded tissues.  What procedures and
	pretreatments are you



	Do You Yahoo!?
	Bid and sell for free at


	Date: 21 Oct 1999 16:01:14 -0500
	From: "Qiu, Yu-Shan" <>
	Subject: Manual for Polycut S

	Can anyone in the histoland kindly send me a copy of Operating
	for Polycut S? Due to the lab moving and people change, we lost ours
	now in need to use this machine to finish our projects. I would
	appreciate your help!

	Yu-Shan Qiu
	Dept. of research
	Shriners Hospital for Children
	Tampa, FL 33617
	Tel. (813)972-2250 Ext.7582
	Fax (813)975-7127


	Date: 21 Oct 1999 17:30:29 -0500
	From: Histonet <>
	Subject: RE: CD31 and CD10

	Dear Dave,

	DAKO Corporation sells a CD31, clone JC/70A (M0823), that can be
used on
	FFPE sections. We recommend enzymatic digestion as a pretreatment.
	Unfortunately, our CD10 can only be used on frozen sections. Please
	Technical Services if you would like further information.

	Danielle McCombs
	DAKO Corporation
	Technical Service Specialist
	(800)235-5743 ext. 5321 

	- -----Original Message-----
	From: Dave Tacha []
	Sent: Thursday, October 21, 1999 11:50 AM
	Subject: CD31 and CD10

	Could some tell me a good CD31 and CD10 antibody  for formalin-fixed
	paraffin embedded tissues.  What procedures and pretreatments are



	Date: 21 Oct 1999 18:45:25 -0500
	From: "Vicki L McKaughan" <>
	Subject: immunostainer

	I am interested in purchasing an automated immunostainer.
currently, I am
	demoing the
	Ventana.  I am schduled to demo the biogenics
	and dako stainers also.  i would appreciated hearing from anyone who
	uses one of these instruments. 

	vicki mckaughan, HT

	What are you N2?  Choose from 150 free e-mail addresses.


	Date: 21 Oct 1999 19:30:17 -0500
	Subject: Glycerin Antigen Retrieval Solution

	A little while ago someone mentioned an antigen retrieval method
using a 
	Glycerin solution for ER-PR.  Could you please mention the specifics
one more 
	time? Thanks


	Date: 21 Oct 1999 22:15:35 -0500
	Subject: Fetus/POC Policy

	Hello Histonetters, 
	               I have an interesting story/question. Twice this year
I have 
	been asked by the family of a patient who had fetal demise
(delivered or 
	c/s), at 19wks to view the fetus. Now, I would normally have no
	(even though I feel it isn't something wonderful to see for the
	family) but in both of these instances, we received the
	already in 10% Zinc Formalin (they have Formalin in the OB unit and
bring the 
	specimens to the lab already submerged). Does anyone out there have
	implemented a policy regarding this situation? My risk manager was
at a loss 
	as to what to come of the situation, so we tried to sway the
	descision (at no avail) and we ended up letting them "view" the
fetus through 
	the original container (frosted and impossible to see anything
	telling them they were unable to open the container due to the
	materials contained in it. This apparently was enough "closure" for
them and 
	they were satisfied. But my risk manager and I still want to make a
	policy concerning this issue. I am in Florida and I was wondering if
state or 
	local laws regarding Formalin and patient/non-hospital employees
	would be an issue? OSHA maybe? We have a release form for fetus' to
	homes only, but we never release Formalin contaminated
tissue/objects to a 
	patient. Has this happened to anyone else out there? What did you
do? I would 
	really appreciate any information you might have. 

	K. Zajic 
	Histology Supervisor
	West Palm Beach, FL

	Here are the messages received yesterday!

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