Have you tried any other clones? Can you get hold of some (free / cheap /
small) samples? I have found that our Ventana Benchmark doesn't like the
occasional clone, ie 1D5 (Ors), and certain clones for CD138 and calretinin.
Jacqui

-----Original Message-----
From: histonet-request@lists.utsouthwestern.edu
[mailto:histonet-request@lists.utsouthwestern.edu] 
Sent: 15 July 2005 18:05
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 20, Issue 17

Send Histonet mailing list submissions to
	histonet@lists.utsouthwestern.edu

To subscribe or unsubscribe via the World Wide Web, visit
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When replying, please edit your Subject line so it is more specific than
"Re: Contents of Histonet digest..."


Today's Topics:

   1. microwave tissue processors (Pereira, Laurie )
   2. Re: low temp thermometer (RCHIOVETTI@aol.com)
   3. Re: low temp thermometer (RCHIOVETTI@aol.com)
   4. Re: low temp thermometer (Karen  Weidenheim)
   5. Re: Paraffin in cassette holding chamber? (Jackie M O'Connor)
   6. Re: Paraffin in cassette holding chamber? (Linda Blazek)
   7. Re: low temp thermometer (Philip Oshel)
   8. Thanks! (wen eng)
   9. RE: overdecalcified bone (Goodwin, Diana)
  10. RE: Paraffin in cassette holding chamber? (Horn, Hazel V)
  11. RE: Paraffin in cassette holding chamber? (Bonnie Whitaker)
  12. Bcl10 (Megan Clarke)
  13. Fwd: CAP (Megan Clarke)
  14. Ventana Benchmark XT and ALK 1 (Malam Jacqueline)
  15. RE: Paraffin in cassette holding chamber? (Molinari, Betsy)
  16. Archiving of Histology Samples in Research (Dimaano, Nena)
  17. RE: Ventana Benchmark XT and ALK 1 (Drew Sally A.)
  18. Re: ADG & Dysferlin troubles  (Jennifer Hofecker)
  19. Xyless? (cfockler@mail1.vcu.edu)


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

Message: 1
Date: Thu, 14 Jul 2005 10:18:42 -0700
From: "Pereira, Laurie " 
Subject: [Histonet] microwave tissue processors
To: 
Message-ID:
	

	
Content-Type: text/plain;	charset="iso-8859-1"

Hello Everyone,

We are a small Veterinary Pathology laboratory and our Pathologist would
like to look into a Microwave Tissue Processor.  He would like to know the
pros and cons of such a processor.  We would only need a 30 specimen
processor.  How long do the tissues need to fix in formalin before using the
microwave?  Does the specimen need to be fresh or can it be 3 days up to 2
weeks old possibly autolyzed?  Does the processor use more alcohols?  We
would appreciate any feedback from techs that have had experience with using
the microwave processors.  Thank you in advance.

Laurie L. Pereira, RVT
SDCADDL (County Veterinarian)
5555 Overland Avenue Suite 4103
San Diego, CA. 92123
Phone: 858-694-3384
Fax: 858-571-4268




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

Message: 2
Date: Thu, 14 Jul 2005 13:34:56 EDT
From: RCHIOVETTI@aol.com
Subject: Re: [Histonet] low temp thermometer
To: sjchtascp@yahoo.com, Histonet@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; charset="US-ASCII"

In a message dated 7/14/2005 9:09:29 AM US Mountain Standard Time,
sjchtascp@yahoo.com writes:

> We're doing allot of liquid nitrogen/isopentane snap freezing and I'd 
> like to purchase a good low temp thermometer.  Any suggestions or 
> advise out there?
> 

Steve,

I've worked in several low-temp "bio"- type labs, and they all used Fluke
digital thermometers.  

You can match the thermocouple to the temperature range you need to measure.

The website can steer you to a dealer or someone for tech questions, but I
know the J- and K-type thermocouples can read temps down near -200 degrees
C.  
That's around the temp of liquid nitrogen (-196) so it should do you well.

Go to Fluke's homepage here.  Then follow the links:
Products -> thermometers -> Fluke 50 Series II Thermometers ->
Specifications

Cheers,

Bob

Robert (Bob) Chiovetti, Ph.D.
The Microscope Works
132 North Elster Drive
Tucson, AZ 85710-3212 USA
Tel./Fax 520-546-4986
Member, Arizona Small Business Association - ASBA


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

Message: 3
Date: Thu, 14 Jul 2005 13:39:08 EDT
From: RCHIOVETTI@aol.com
Subject: Re: [Histonet] low temp thermometer
To: RCHIOVETTI@aol.com, sjchtascp@yahoo.com,
	Histonet@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; charset="US-ASCII"

In a message dated 7/14/2005 10:36:00 AM US Mountain Standard Time,
RCHIOVETTI@aol.com writes:

> Go to Fluke's homepage here.  Then follow the links:
> Products -> thermometers -> Fluke 50 Series II Thermometers -> 
> Specifications
> 
> 

Oops, sorry, can't post an active link...so...Fluke's homepage for the USA
is:



Bob

Robert (Bob) Chiovetti, Ph.D.
The Microscope Works
132 North Elster Drive
Tucson, AZ 85710-3212 USA
Tel./Fax 520-546-4986
Member, Arizona Small Business Association - ASBA


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

Message: 4
Date: Thu, 14 Jul 2005 13:39:26 -0400
From: "Karen  Weidenheim" 
Subject: Re: [Histonet] low temp thermometer
To: , 
Message-ID: 
Content-Type: text/plain; charset=US-ASCII

Dear Steve, I presume you are freezing muscle biopsies, or possibly research
material.
This isopentane/liquid nitrogen method is more an art than a science. 
Altitude of your lab and humidity of your ambient air, are the 2 major
limiting factors.  The most important thing in the freezing is not measuring
the temperature but rather visual assessment of the state of the isopentane.
We put the isopentane in a 250 ml stainless steel beaker and lower it slowly
into the liquid nitrogen.  We wait to freeze, until the isopentane has
formed ice all around the walls of the beaker and all the way up the wall of
the beaker, and completely across the bottom of the beaker.  It will be
syrupy.  We use 25 seconds of freezing (I know the books say 5-10 seconds).
Our lab is at sea level.  It is also often humid here.  We discard the
isopentane every 3 months when there is no major humidity and more often
(every 6 weeks) when we have a run of humid weather, or when we have a
problem with freezing artifact, or when I think my assistant has been sloppy
with its storage.
We keep the beaker very clean i.e. we scour with steel wool.  We make sure
there is no residual water in the beaker (I keep 2 beakers so one is freshly
clean and dry at all times).
I have not used the thermometer since I was a young attending a long time
ago!!!
Best
Karen

Karen M. Weidenheim, M.D.
Professor of Pathology, Clinical Neurology and Clinical Neurosurgery Albert
Einstein College of Medicine Chief, Division of Neuropathology Montefiore
Medical Center
111 East 210th Street
Bronx, NY   10467 
(718) 920-4446
FAX (718) 653-3409
Beeper (917) 556 3696
CONFIDENTIALITY NOTICE:  This email, including any attachments, is for the
sole use of the intended recipient(s).  The information contained in this
message may be private and confidential, and may also be subject to the work
product doctrine.  Any unauthorized review, use, disclosure or distribution
is prohibited.  If you are not the intended recipient, please contact the
sender by reply e-mail and destroy all copies of the original message.
Thank you.

>>> Steven Coakley  7/14/2005 12:07:15 PM >>>
We're doing allot of liquid nitrogen/isopentane snap freezing and I'd like
to purchase a good low temp thermometer.  Any suggestions or advise out
there?
 
Steve

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

Message: 5
Date: Thu, 14 Jul 2005 12:58:14 -0500
From: "Jackie M O'Connor" 
Subject: Re: [Histonet] Paraffin in cassette holding chamber?
To: Michele_Marggi@ssmhc.com
Cc: histonet@lists.utsouthwestern.edu
Message-ID:
	

	
Content-Type: text/plain; charset="us-ascii"

I've done this both ways, with and without.  I once thought that the
cassettes had to be held in molten paraffin to ensure a homogenous block - 

I've since changed my mind. It doesn't matter.  I prefer a hot dry holding
chamber - less messy. 

Jacqueline M. O'Connor HT(ASCP) QIHC
Assistant Scientist
Discovery Cancer R4N2 AP3
Jackie.O'Connor@abbott.com





Michele_Marggi@ssmhc.com
Sent by: histonet-bounces@lists.utsouthwestern.edu
07/14/2005 11:47 AM

 
        To:     histonet@lists.utsouthwestern.edu
        cc: 
        Subject:        [Histonet] Paraffin in cassette holding chamber?


I want to get an idea of who is (and who is not, and why) adding paraffin
to the cassette holding chamber of the embedding center.  Once
processed-How long are tissues ok to sit without paraffin on them?

All thoughts, opinions, experience is appreciated...Thanks.

Michele Marggi, HT
Surgical Pathology Supervisor
St. Marys Hospital Medical Center
707 S Mills Street
Madison WI  53715
Telephone: 608.258.6930
Fax: 608.258.6268


-----------------------------------------
Confidentiality Notice: This email message, including any attachments, is
for the sole use of the intended recipient(s) and may contain confidential
and privileged information. Any unauthorized review, use, disclosure or
distribution is prohibited. If you are not the intended recipient, please
contact the sender by reply email and destroy all copies of the original
message.


_______________________________________________
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Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet




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

Message: 6
Date: Thu, 14 Jul 2005 14:05:01 -0400
From: "Linda Blazek" 
Subject: Re: [Histonet] Paraffin in cassette holding chamber?
To: histonet@lists.utsouthwestern.edu, Michele_Marggi@ssmhc.com
Message-ID: 
Content-Type: text/plain; charset=US-ASCII

I've done this both with and without melted paraffin.  I prefer having a dry
holding tank.  Less mess.  Less waste of paraffin also.   As long as the
holding tank keeps the cassettes from solidifying I think they can sit for
quite awhile.  Or at least as long as it take to embed a tank full.
 
 
Linda  Blazek, HT (ASCP)
Department of Pathology
Children's Medical Center
Dayton, Ohio  45404
(937) 641-3358
fax (937)641-5482
blazekl@childrensdayton.org


>>>  07/14/2005 12:47 PM >>>

I want to get an idea of who is (and who is not, and why) adding paraffin
to the cassette holding chamber of the embedding center.  Once
processed-How long are tissues ok to sit without paraffin on them?

All thoughts, opinions, experience is appreciated...Thanks.

Michele Marggi, HT
Surgical Pathology Supervisor
St. Marys Hospital Medical Center
707 S Mills Street
Madison WI  53715
Telephone: 608.258.6930
Fax: 608.258.6268


-----------------------------------------
Confidentiality Notice: This email message, including any attachments, is
for the sole use of the intended recipient(s) and may contain confidential
and privileged information. Any unauthorized review, use, disclosure or
distribution is prohibited. If you are not the intended recipient, please
contact the sender by reply email and destroy all copies of the original
message.


_______________________________________________
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------------------------------

Message: 7
Date: Thu, 14 Jul 2005 13:29:52 -0500
From: Philip Oshel 
Subject: Re: [Histonet] low temp thermometer
To: Histonet@Pathology.swmed.edu
Message-ID: 
Content-Type: text/plain; format=flowed; charset=us-ascii

You discard your isopentane every 3 months? Is this stuff you've been 
using for freezing, and you keep using the same isopentane? And it 
stays cold the whole time?
I hope not.
The cold isopentane is getting enriched in oxygen as it's being used, 
and this heightens the fire hazard.
Why not freeze into (all right, everyone who's read my rants before 
can ignore me now) slush nitrogen? Colder, faster freezing, better 
freezing, and no flammable/explosive liquids or gases involved.

Phil

>Dear Steve, I presume you are freezing muscle biopsies, or possibly
>research material.
>This isopentane/liquid nitrogen method is more an art than a science.
>Altitude of your lab and humidity of your ambient air, are the 2 major
>limiting factors.  The most important thing in the freezing is not
>measuring the temperature but rather visual assessment of the state of
>the isopentane.  We put the isopentane in a 250 ml stainless steel
>beaker and lower it slowly into the liquid nitrogen.  We wait to freeze,
>until the isopentane has formed ice all around the walls of the beaker
>and all the way up the wall of the beaker, and completely across the
>bottom of the beaker.  It will be syrupy.  We use 25 seconds of freezing
>(I know the books say 5-10 seconds).
>Our lab is at sea level.  It is also often humid here.  We discard the
>isopentane every 3 months when there is no major humidity and more often
>(every 6 weeks) when we have a run of humid weather, or when we have a
>problem with freezing artifact, or when I think my assistant has been
>sloppy with its storage.
>We keep the beaker very clean i.e. we scour with steel wool.  We make
>sure there is no residual water in the beaker (I keep 2 beakers so one
>is freshly clean and dry at all times).
>I have not used the thermometer since I was a young attending a long
>time ago!!!
>Best
>Karen
>
>Karen M. Weidenheim, M.D.
>Professor of Pathology, Clinical Neurology and Clinical Neurosurgery
>Albert Einstein College of Medicine
>Chief, Division of Neuropathology
>Montefiore Medical Center
>111 East 210th Street
>Bronx, NY   10467
>(718) 920-4446
>FAX (718) 653-3409
>Beeper (917) 556 3696
>CONFIDENTIALITY NOTICE:  This email, including any attachments, is for
>the sole use of the intended recipient(s).  The information contained in
>this message may be private and confidential, and may also be subject to
>the work product doctrine.  Any unauthorized review, use, disclosure or
>distribution is prohibited.  If you are not the intended recipient,
>please contact the sender by reply e-mail and destroy all copies of the
>original message.  Thank you.
>
>>>>  Steven Coakley  7/14/2005 12:07:15 PM >>>
>We're doing allot of liquid nitrogen/isopentane snap freezing and I'd
>like to purchase a good low temp thermometer.  Any suggestions or advise
>out there?
>
>Steve
>
>
>---------------------------------
>  Start your day with Yahoo! - make it your home page
>_______________________________________________
>Histonet mailing list
>Histonet@lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>_______________________________________________
>Histonet mailing list
>Histonet@lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet

-- 
Philip Oshel
Supervisor, BBPIC microscopy facility
Department of Animal Sciences
University of Wisconsin
1675 Observatory Drive
Madison,  WI  53706
voice: (608) 263-4162
fax: (608) 262-5157 (dept. fax)
http://www.ansci.wisc.edu/microscopy.htm



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

Message: 8
Date: Thu, 14 Jul 2005 11:30:37 -0700 (PDT)
From: wen eng 
Subject: [Histonet] Thanks!
To: histonet 
Message-ID: <20050714183037.83786.qmail@web53403.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Thanks to all who answered my question about H&E after IHC. I really
appreciate the help I got from you guys. Have a nice day!
 
Wen

__________________________________________________
Do You Yahoo!?
Tired of spam?  Yahoo! Mail has the best spam protection around 
http://mail.yahoo.com 

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

Message: 9
Date: Thu, 14 Jul 2005 14:50:36 -0400
From: "Goodwin, Diana" 
Subject: RE: [Histonet] overdecalcified bone
To: 'Hermina Borgerink' 
Cc: "Histonet \(E-mail\)" 
Message-ID:
	<992899E9EC268548AB8DDE246AF88473055F5251@PAHEX01.uphs.upenn.edu>
Content-Type: text/plain;	charset="iso-8859-1"

Hermina:
 
We used the 5% periodic acid overnight method with remarkable results.  I am
writing a procedure...do you have any idea why these methods work, ie, the
chemistry behind the method?  I found the original method in the AFIP
manual, but no theory was given.
 
Thanks,
Diana G. Goodwin
Department of Pathology
Pennsylvania Hospital
800 Spruce St., Preston 655-C
Philadelphia, PA  19107
 
ph:  215-829-6532
fax:  215-829-7564
e-mail:  goodwind@pahosp.com

----Original Message-----
From: Hermina Borgerink [mailto:hborgeri@wfubmc.edu]
Sent: Wednesday, July 13, 2005 1:40 PM
To: Goodwin, Diana; kalschev@svm.vetmed.wisc.edu
Cc: Histonet (E-mail)
Subject: RE: [Histonet] overdecalcified bone



>From Theory and practice of Histotechnology, Sheehan and Hrapchak, Appendix
B, p. 445.  

 

To restore basophilic properties to tissues as the result of overexposure to
decalcifying solution:

 

Method 1

1.    Deparaffinize and hydrate slides

2.    Place slides in 5% aqueous sodium bicarbonate solution from 4 hours to
overnight

3.    Wash in tap water for 5 minutes

4.    Stain as desired

 

Method 2

In step 2, use 5% aqueous solution of ammonium sulfide overnight

 

Method 3

In step 2, use 5% aqueous solution of periodic acid overnight

 

Hopefully one of these methods will work for you.

Hermina

 

 

 

Hermina M. Borgerink, BA, HTL(ASCP)QIHC

Wake Forest University Health Sciences

Department of Pathology

Medical Center Blvd.

Winston-Salem, NC 27157

Tel. (336) 716-1538

Fax. (336) 716-1515

e-mail hborgeri@wfubmc.edu 

 

"Treat a man as he appears to be, and you make him worse.  But treat a man
as if he already were what he potentially could be, and you make him what he
should be."  (Goethe)

 

This electronic message, including any attachments, is confidential and
proprietary and is solely for the intended recipient.  If you are not the
intended recipient, this message was sent to you in error and you are hereby
advised that any review, disclosure, copying, distribution or use of this
message, or any of the information included therein, is unauthorized and
strictly prohibited.  If you have received this electronic transmission in
error, please immediately notify the sender by reply and permanently delete
all copies of this message and its attachments.  Thank you. 

 

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Goodwin,
Diana
Sent: Wednesday, July 13, 2005 1:12 PM
To: 'kalschev@svm.vetmed.wisc.edu'
Cc: Histonet (E-mail)
Subject: [Histonet] overdecalcified bone

 

Hi, Vicki.

 

Is there a procedure to restore the nuclear staining uptake in

over-decalcified human bone?

 

Diana Goodwin

Supervisor, Anatomic Pathology

Pennsylvania Hospital, Preston 655-C

 

ph:  215-829-6532

fax:  215-829-7564

e-mail:  goodwind@pahosp.com

 

_______________________________________________

Histonet mailing list

Histonet@lists.utsouthwestern.edu

http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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

Message: 10
Date: Thu, 14 Jul 2005 14:32:44 -0500
From: "Horn, Hazel V" 
Subject: RE: [Histonet] Paraffin in cassette holding chamber?
To: Michele_Marggi@ssmhc.com,	histonet@lists.utsouthwestern.edu
Message-ID:
	<9AE8AA9E1F644B4AA6C155FB6FD51C63038BDDA0@EMAIL.archildrens.org>
Content-Type: text/plain; charset=us-ascii

We do not add paraffin to our holding chamber.   But they do not sit
there very long, as we start embedding right away.   Is there a reason
you want them to sit for a time before embedding? 


Hazel Horn, HT/HTL (ASCP)
Histology Supervisor
Arkansas Children's Hospital - Changing Children's Lives

Phone - 501.364.4240
Fax - 501.364.3912 

Visit us on the web at      www. archildrens.org


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Michele_Marggi@ssmhc.com
Sent: Thursday, July 14, 2005 11:48 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Paraffin in cassette holding chamber?

I want to get an idea of who is (and who is not, and why) adding
paraffin to the cassette holding chamber of the embedding center.  Once
processed-How long are tissues ok to sit without paraffin on them?

All thoughts, opinions, experience is appreciated...Thanks.

Michele Marggi, HT
Surgical Pathology Supervisor
St. Marys Hospital Medical Center
707 S Mills Street
Madison WI  53715
Telephone: 608.258.6930
Fax: 608.258.6268


-----------------------------------------
Confidentiality Notice: This email message, including any attachments,
is for the sole use of the intended recipient(s) and may contain
confidential and privileged information. Any unauthorized review, use,
disclosure or distribution is prohibited. If you are not the intended
recipient, please contact the sender by reply email and destroy all
copies of the original message.


_______________________________________________
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Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


----------------------------------------------------------------------------
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The information contained in this message may be privileged and confidential
and protected from disclosure. If the reader of this message is not the
intended recipient, or an employee or agent responsible for delivering this
message to the intended recipient, you are hereby notified that any
dissemination, distribution or copying of this communication is strictly
prohibited. If you have received this communication in error, please notify
us immediately by replying to the message and deleting it from your
computer. Thank you. 
============================================================================
==




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

Message: 11
Date: Thu, 14 Jul 2005 14:42:53 -0500
From: "Bonnie Whitaker" 
Subject: RE: [Histonet] Paraffin in cassette holding chamber?
To: "'Horn, Hazel V'" ,
	,	
Message-ID: <000001c588ac$3b8fcf00$3601a8c0@brownpathology.net>
Content-Type: text/plain;	charset="us-ascii"

If it's going to be a really long time (or even to hold extra tissue,
unembedded for a great length of time) just pull the cassettes out and then
put them back in to heat up right before embedding.

Bonnie Whitaker


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel
V
Sent: Thursday, July 14, 2005 2:33 PM
To: Michele_Marggi@ssmhc.com; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] Paraffin in cassette holding chamber?


We do not add paraffin to our holding chamber.   But they do not sit
there very long, as we start embedding right away.   Is there a reason
you want them to sit for a time before embedding? 


Hazel Horn, HT/HTL (ASCP)
Histology Supervisor
Arkansas Children's Hospital - Changing Children's Lives

Phone - 501.364.4240
Fax - 501.364.3912 

Visit us on the web at      www. archildrens.org


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Michele_Marggi@ssmhc.com
Sent: Thursday, July 14, 2005 11:48 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Paraffin in cassette holding chamber?

I want to get an idea of who is (and who is not, and why) adding paraffin to
the cassette holding chamber of the embedding center.  Once processed-How
long are tissues ok to sit without paraffin on them?

All thoughts, opinions, experience is appreciated...Thanks.

Michele Marggi, HT
Surgical Pathology Supervisor
St. Marys Hospital Medical Center
707 S Mills Street
Madison WI  53715
Telephone: 608.258.6930
Fax: 608.258.6268


-----------------------------------------
Confidentiality Notice: This email message, including any attachments, is
for the sole use of the intended recipient(s) and may contain confidential
and privileged information. Any unauthorized review, use, disclosure or
distribution is prohibited. If you are not the intended recipient, please
contact the sender by reply email and destroy all copies of the original
message.


_______________________________________________
Histonet mailing list
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet


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The information contained in this message may be privileged and confidential
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------------------------------

Message: 12
Date: Fri, 15 Jul 2005 12:01:38 +1000
From: "Megan Clarke" 
Subject: [Histonet] Bcl10
To: 
Message-ID: 
Content-Type: text/plain; charset=US-ASCII

Hello Histonetters,
We would like to see if anyone can help us with an antibody optimisation for
Bcl10?
We have the Zymed Bcl 10 ,clone 151.
We have tried HIER using citrate ph 6.0,citrate EDTA ph 8.0,and Dako TUR
both on the Ventana and the Bond Max.Our dilution ranges have been from a 1
in 50 up to 1 in 2000.
Our slides look "muddy" with distorted morphology.With no specific
staining.We are using a reactive tonsil as our control.
If anyone is familiar using this and can offer us some help we would really
appreciate it.
Thank you very much
Megan Clarke and Zenobia Haffajee
Immunohistochemisty Lab
HAPS>JHH
Newcastle
AUSTRALIA




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

Message: 13
Date: Fri, 15 Jul 2005 12:11:54 +1000
From: "Megan Clarke" 
Subject: [Histonet] Fwd: CAP
To: 
Message-ID: 
Content-Type: text/plain; charset=US-ASCII



>>> Megan Clarke 07/15/05 12:10 PM >>>
Hello Hitonetters,
Could anyone please send me some information regarding you Quality Control
Program on Immunohistochemisrty?
I think it is called CAP?? 
Any information would be greatly appreciated.
Thanks
Megan Clarke
Immunohistochemistry Lab
HAPS.JHH
Newcastle AUSTRALIA




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

Message: 14
Date: Fri, 15 Jul 2005 09:20:43 +0100
From: Malam Jacqueline 
Subject: [Histonet] Ventana Benchmark XT and ALK 1
To: "Histonet submissions (histonet@lists.utsouthwestern.edu)"
	
Message-ID: 
Content-Type: text/plain

Does anyone out there do ALK 1 on the Ventana Benchmark? If so, I would be
grateful if you could let me know what clone you use, what the protocol and
dilution are, and if you could spare me a positive control block. 
I have tried 2 clones - Dako's ALK 1 and Lab Vision's SP8 but to no avail; I
have had experience of certain clones of some antisera (Vector's CD 138 and
Dako's calretinin for example) not working on the Benchmark and it's
possible that these clones are not suitable. Having said that, we only had 2
cases and possibly they may have been negative for ALK 1 anyway. I used the
lowest recommended dilution, both the mild and standard heat protocols, and
the 3 primary incubation temperatures (room, 37 and 42) and even tried
protease and no retrieval but got nowhere. I have also tried to get a known
positive control block but no one I've tried has one!
All assistance appreciated!
Jacqui Malam
Royal Infirmary
Lancaster
England



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

Message: 15
Date: Fri, 15 Jul 2005 05:48:48 -0500
From: "Molinari, Betsy" 
Subject: RE: [Histonet] Paraffin in cassette holding chamber?
To: 
Message-ID: 
Content-Type: text/plain;	charset="us-ascii"

I do not put any extra paraffin into the holding chamber. If I am not
going to embed right away I take the cassettes out of the processor and
let them cool off with the paraffin hardening around it. I have not had
any complaints with tissue handled in this way.
Betsy Molinari HT(ASCP)
Texas Heart Institute
Cardiovascular Pathology
6770 Bertner Avenue
Houston, TX 77030
832-355-6524
832-355-6812 ( fax)

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
Michele_Marggi@ssmhc.com
Sent: Thursday, July 14, 2005 11:48 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Paraffin in cassette holding chamber?

I want to get an idea of who is (and who is not, and why) adding
paraffin
to the cassette holding chamber of the embedding center.  Once
processed-How long are tissues ok to sit without paraffin on them?

All thoughts, opinions, experience is appreciated...Thanks.

Michele Marggi, HT
Surgical Pathology Supervisor
St. Marys Hospital Medical Center
707 S Mills Street
Madison WI  53715
Telephone: 608.258.6930
Fax: 608.258.6268


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

Message: 16
Date: Fri, 15 Jul 2005 08:34:16 -0400
From: "Dimaano, Nena" 
Subject: [Histonet] Archiving of Histology Samples in Research
To: 
Message-ID:
	
<20BE9059B6FC2D4B9E8FF2A9E2C4EEBD12A4F837@HOS2KEXCHCL.howost.strykercorp.com
>
	
Content-Type: text/plain;	charset="iso-8859-1"

Hi All,
 I was wondering if you guys can help me answer the following questions
about archiving histology samples.
Our Histology Lab archive the samples(including wet samples stored in 10%
formalin, blocks both paraffin and plastics)  indefinitely and since we are
running out of space it is becoming a great deal of problem. 
 
The following are my questions:

*	How long do you keep the samples stored in 10% formalin? 

*	How long do you store the blocks (paraffin and plastics) in
research? 

*	Is there any agencies like the FDA, CDC or CLIA outlining this task?

thanks,
Nena
 
Nena Dimaano
Orthobiologics, R&D
Stryker Orthopaedics
325 Corporate Drive
Mahwah, NJ 07430
tel: 201-831-5338
fax: 201-831-6224
email: Nena.Dimaano@stryker.com
 
 


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

Message: 17
Date: Fri, 15 Jul 2005 08:14:34 -0500
From: "Drew Sally A." 
Subject: RE: [Histonet] Ventana Benchmark XT and ALK 1
To: "Histonet \(Histonet\)" 
Message-ID:
	

Content-Type: text/plain;	charset="us-ascii"

My co-worker just successfully ran a predilute polyclonal ALK from
BioCare Medical on our Benchmark with an MCC1, 32"ttn and no  block.  We
had previously only run it on the Nexes and haven't run it on our XT
yet.  
Unfortunately, we are not flush w/ control tissue (we only have 1
positive case from 1999) so I can't send you a block.  However, would a
few cut slides help you get started?
I know that when I first worked up this antibody I didn't have much luck
w/ the rabbit monoclonals I tried.

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malam
Jacqueline
Sent: Friday, July 15, 2005 3:21 AM
To: Histonet submissions (histonet@lists.utsouthwestern.edu)
Subject: [Histonet] Ventana Benchmark XT and ALK 1


Does anyone out there do ALK 1 on the Ventana Benchmark? If so, I would
be grateful if you could let me know what clone you use, what the
protocol and dilution are, and if you could spare me a positive control
block. 
I have tried 2 clones - Dako's ALK 1 and Lab Vision's SP8 but to no
avail; I have had experience of certain clones of some antisera
(Vector's CD 138 and Dako's calretinin for example) not working on the
Benchmark and it's possible that these clones are not suitable. Having
said that, we only had 2 cases and possibly they may have been negative
for ALK 1 anyway. I used the lowest recommended dilution, both the mild
and standard heat protocols, and the 3 primary incubation temperatures
(room, 37 and 42) and even tried protease and no retrieval but got
nowhere. I have also tried to get a known positive control block but no
one I've tried has one! All assistance appreciated! Jacqui Malam Royal
Infirmary Lancaster England



DISCLAIMER: This e-mail is confidential and privileged. If you are not
the intended recipient please accept our apologies; please do not
disclose, copy or distribute information in this e-mail or take any
action in reliance on its contents: to do so is strictly prohibited and
may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this
message has gone astray before deleting it.  Comments or opinions
expressed in this email are those of their respective contributors only.
The views expressed do not represent the views of the Trust, its
management or employees. Morecambe Bay Hospitals NHS Trust is not
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comments written within.Thank you for your co-operation.


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

Message: 18
Date: Fri, 15 Jul 2005 06:32:51 -0700 (PDT)
From: Jennifer Hofecker 
Subject: [Histonet] Re: ADG & Dysferlin troubles 
To: histonet@lists.utsouthwestern.edu, LewisS@ccri.net
Message-ID: <20050715133251.93873.qmail@web33506.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Hi Everyone, sorry for the delay in responding, my
histonet digest was "lost in space" for a few days. 
We don't do ADG here, but I also use the Dysferlin
antibody from Novacastra (purchased from vector) with
regular IHC (no FITC) for frozen sections. I use Clone
Ham1/7B6(new vector number VP-D503).  
  Air dry frozen sections (usually 20 min or more). 
Dilution:  1:20 at rm temp 1 hr.
 I use with Vector R.T.U. vectastain kit (use
secondary 30 min@37degrees C. ABC reagent 1hr. rm
temp.  I use DAB from Vector for 15 min.  I usually do
a very pale counterstain or none at all depending on
pathologist.
I hope this is of some help to you.
Have a good weekend,

Jennifer Hofecker
Nashville,TN

Date: Wed, 13 Jul 2005 07:43:36 -0400
From: Dana Settembre 
Subject: Re: [Histonet] ADG & Dysferlin troubles
To: LewisS@ccri.net, histonet@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; charset=US-ASCII

Sarah,
I use  Dysferlin from Novocastra on frozen sections,
but not FITC, just
regular IHC. If you're desperate we can talk.
I use it with a detection kit for mouse, any vendor
will usually do and
DAB as the chromogen. No special scope is needed.
Dana Settembre
University Hospital-UMDNJ
Newark, NJ

>>> "Lewis, Sarah"  7/12/2005 1:48:17
PM >>>
Hello histoneters,
 
    I could really use any procedural information
anyone has on
Alpha-dystroglycan (antibody only available thru
Upstate) and Dysferlin
(antibody from VectorLabs).
 
        I have been attempting the stains for 2months
now with no
satisfying results. I have tried several different
procedures/fixatives/incubation times/dilutions/. I
perform a battery 
of
14 neuromuscular Immunoflourescence, they are all
BEAUTIFUL fitc green
with no background staining (picture perfect)
.....except these two
boogers. PLEASE HELP me make my pathologist proud!!!
ALL THE CREDIT TO
HISTONET!!!!   

Sarah E. Lewis 
Histotechnician 
Childrens Research 
Center for Gene Therapy 
700 Childrens Dr Rm WA3112 
Columbus OH 43205 
(614)-722-2204 
LewisS@ccri.net 

 


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

Message: 19
Date: Fri, 15 Jul 2005 09:38:00 -0400
From: 
Subject: [Histonet] Xyless?
To: Amos Brooks 
Cc: histonet@lists.utsouthwestern.edu, Rcartun@harthosp.org,
	kappeler@patho.unibe.ch
Message-ID: <200507151338.JAA30123@despina.vcu.edu>
Content-Type: text/plain; charset=iso-8859-1

-------------------
> Rich & Andi,
>     Rich, I'd like to second Andi's recommendation for Sigma as a
source 
> for Fast Myosin although we don't bother with HIER. Give it a whirl
both 
> ways, you never know her's might be better :-)
>     Andi, while you asked Rich I figured I'd chime in for Dako as a 
> source for Hepatitis (core & surface). Unfortunately I'm out of the
lab 
> right now so I don't have the cat #s. If  you need them just drop me
a 
> line and I'll rummage them up.
> Good luck guys,
> Amos
> 
>
//////////////////////////////////////////////////////////////////////
////////////////
> 
> Message: 22
> Date: Tue, 12 Jul 2005 16:37:08 +0200
> From: "Andi Kappeler" 
> Subject: Re: [Histonet] Fast myosin IHC
> To: "Richard Cartun" ,	"Histonet"
> 	
> Message-ID: <008801c586ef$40bf45f0$27955c82@patho.unibe.ch>
> Content-Type: text/plain; format=flowed; charset="iso-8859-1";
> 	reply-type=original
> 
> Hi Richard
> 
> We successfully use:
> - mo-a-Myosin II (fast), clone MY-32, Sigma M 4276, working conc.
approx 10 
> ug Ig/ml (corresponds to 1:400 with the lot we have), HIER in
citrate pH 6.0 
> (pressure cooker)
> - mo-a-Myosin I (slow), clone NOQ7.5.4D, Sigma M 8421, working conc.
approx 
> 7.5 ug Ig/ml (corresponds to 1:1000 with the lot we have), HIER in
citrate 
> pH 6.0 (pressure cooker)
> Both mAbs work very well (see 
> http://www.pathology.unibe.ch/Diagn/speztech/spez_ihc.htm for a pic
(german 
> website, sorry).
> 
> Can you give me a hint in return, where you get your antibodies
against 
> Hepatitis B surface Ag (HBs) and core Ag (HBc) from? Our source is
no longer 
> able to supply them, due to some bureaucratic / eurocratic rules,
now I'm 
> looking for a reliable replacement.
> Many thanks in advance!
> 
> Best regards
> Andi Kappeler
> Institute of Pathology, University of Bern, Switzerland
> 
> 
> 
> 
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> Hello All . .. I just recently moved to a lab that uses a few
changes of Xyless, (xylene substitute) in their stainer and processor.
 .can anyone give me any information about this product? I can't find
anything useful on the web.  Thanks in advance!


Candyce LeTellier  
Histotechnologist



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

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End of Histonet Digest, Vol 20, Issue 17
****************************************


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deleting it.  Comments or opinions expressed in this email are those of
their respective contributors only. The views expressed do not represent the
views of the Trust, its management or employees. Morecambe Bay Hospitals NHS
Trust is not responsible and disclaims any and all liability for the content
of comments written within.Thank you for your co-operation.



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