[Histonet] RE: RE: (Histonet) Sakura Auto Stainer (Charles Scouten)

From:"Aldo Anile"

The TST 44 by Medite has a large number reagent stations giving it the ability to stain large numbers of slides from different staining protocols in a short period of time. 

There are 44 stations in the Medite TST 44, 6 of these are water stations, along with 4 load stations, 4 unload stations and 30 staining stations. This number of stations allows for the staining of slides with H&E and PAP simultaneously.

Depending upon your protocols, you will have reagent stations left over for other staining protocols.

Eg.
A standard H&E staining program may only require up to 13 stations. These would include two de-waxing, three re-hydrating, Haematoxylin, Eosin, blueing solution, three dehydrating and two clearing. 

A cytology PAP stain may only require up to 16 solutions. This allows for two initial dehydrating solutions, three stains, Haematoxylin, OG6 and EA50, a blueing solution, two 95% alcohol washes before the OG6 and two 95% alcohol washes between the OG6 and EA50 followed by four dehydrating alcohols and two clearing solutions.

We have a number of users in your exact situation, please fell free to contact me to discuss this further.

Aldo Anile
Application Consultant
HD Scientific

Phone:	+ 61 3 9364 9569
Fax No:	+ 61 3 9364 9479
Mobile: 	0408 471 485
E-mail:	aldo.anile@hdscientific.com.au 
Web:	www.hdscientific.com.au 



-----Original Message-----
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[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of
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Sent: Thursday, 22 July 2004 18:53
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 8, Issue 32


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Today's Topics:

   1. Re: ] Gold Chloride for Retic (Alton D. Floyd)
   2. adhesive tape  & MMA Sections - Turpentine? (Johnson, Teri)
   3. Agarose blocks (Roberta Horner)
   4. RE: Disposing of expired IHC reagents: Survey
      (Poteete, Jacquie A.)
   5. formalin nomenclature (Subratab)
   6. RE: Blood vessel antibodies for rabbit models (Flynn, Evelyn)
   7. Re: Oil of wintergreen (Sarah Jones)
   8. Re: AURANTIA (Sarah Jones)
   9. cryostat sectioning of unfixed human brains (Carl)
  10. RE: Agarose blocks (Charles  Scouten)
  11. RE: DECONTAMINATING CRYOSTATS (Charles  Scouten)
  12. Webmaster attention (Denise Crowley)
  13. RE: DECONTAMINATING CRYOSTATS (Charles  Scouten)
  14. RE: (Histonet) Sakura Auto Stainer (Charles  Scouten)
  15. C4D (james.zimmerman@pharma.novartis.com)
  16. Darkroom Processor (Mitchell (Jean))
  17. hard paraffin (Sarah Jones)
  18. brightfield 3D reconstruction (Connolly, Brett M)
  19. hard paraffin additive (Gayle Callis)
  20. Re: brightfield 3D reconstruction (Gayle Callis)
  21. Re: Blood vessel antibodies for rabbit models (Rocan)
  22. Indirect Immunofluorescence (Brody,Juanita X)
  23. Re: DECONTAMINATING CRYOSTATS (lpwenk@sbcglobal.net)
  24. Pan Melanoma cocktail from "Not" Mouse...?? (Swaram)
  25. Re: hard paraffin additive (Sarah Jones)
  26. RE: Galectin-3 (Brett, Lawrence)
  27. RE: Agarose blocks (Margaret Blount)
  28. RE: brightfield 3D reconstruction (Margaret Blount)


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

Message: 1
Date: Wed, 21 Jul 2004 12:58:11 -0400
From: "Alton D. Floyd" 
Subject: Re: [Histonet] ] Gold Chloride for Retic
To: David.Edmondson@christie-tr.nwest.nhs.uk
Cc: Histonet@lists.utsouthwestern.edu
Message-ID: <20040721.125814.-279241.1.al.floyd@juno.com>
Content-Type: text/plain; charset=us-ascii

Hello David,

The procedure you are remembering is the generation of gold colloids for
the purpose of performing immunostaining on plastic sections for electron
microscopy.  And you are correct, the color of the scattered light is
indicative of the particle size.  Not absolute, since individual eyes
vary in color perception so much.  Actual size generated depends on the
conditions employed in manufacturing the good particles, which are  then
cleaned by ultracentrifugation prior to use.  The colloid itself is
generated by the protein coating (which can contain the primary
antibodies) which is applied to the particles.

Al Floyd
23126 South Shore Drive
Edwardsburg, MI 49112
(269) 699-7182 phone & fax



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

Message: 2
Date: Wed, 21 Jul 2004 12:11:08 -0500
From: "Johnson, Teri" 
Subject: [Histonet] adhesive tape  & MMA Sections - Turpentine?
To: "Histonet" 
Message-ID:
	
Content-Type: text/plain;	charset="us-ascii"

Patsy was describing the technique for using the paraffin tape transfer
system on MMA blocks.  One needs to remove the tape with an aliphatic or
aromatic hydrocarbon before hydration and subsequent staining.  Phillip
was describing tape transfer  method for cryosections.
 

Teri Johnson
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110
tjj@stowers-institute.org


 


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

Message: 3
Date: Wed, 21 Jul 2004 13:39:10 -0400
From: "Roberta Horner" 
Subject: [Histonet] Agarose blocks
To: 
Message-ID: <00a801c46f49$a1dd7f00$8861ba92@padlspsu.psu.edu>
Content-Type: text/plain;	charset="US-ASCII"

I just received tissue in agarose and I tried cutting a couple of them with
very little luck.  I have never worked with this before.  Is there a trick
to sectioning it?  Should it have been processed differently?  I didn't know
what was in the block until I embedded it.

Thank you for any help.

Roberta Horner HT/HTL

 

Animal Diagnostic Lab

Penn State University



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

Message: 4
Date: Wed, 21 Jul 2004 12:37:12 -0500
From: "Poteete, Jacquie A." 
Subject: RE: [Histonet] Disposing of expired IHC reagents: Survey
To: 'Joe Nocito' , Histonet
	
Message-ID:
	
Content-Type: text/plain

Our hospital Safety Department prefers that we use the biohazard waste
system for antibody and detection system reagent disposal.

Jacquie Poteete MT(ASCP)QIHC
Lead Technologist, IHC Laboratory
Saint Francis Hospital, Tulsa, OK
japoteete@saintfrancis.com

> -----Original Message-----
> From:	Joe Nocito [SMTP:JNocito@Pathreflab.com]
> Sent:	Wednesday, July 21, 2004 11:49 AM
> To:	Histonet
> Subject:	[Histonet] Disposing of expired IHC reagents: Survey
> 
> Hey 'netters,
> Need a little info if you please. How are you disposing of expired primary
> antibodies and detection reagents? In the trash? Biohazard? Drain? Big
> bonfire?
> 
> 
> Joe Nocito, BS, HT(ASCP) QIHC
> Histology Manager
> Pathology Reference Lab
> San Antonio, TX
> 
> 
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> 
> 
> 
********* Email Confidentiality Statement ********* 
Visit http://www.saintfrancis.com/emailconf.asp



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

Message: 5
Date: Wed, 21 Jul 2004 23:47:41 +0600
From: Subratab 
Subject: [Histonet] formalin nomenclature
To: 
Message-ID: <200407211749.i6LHnaCa027768@mailout.proshikanet.com>
Content-Type: text/plain; charset="iso-8859-1";

Dear all
Here is a nice letter regarding the nomenclature of formalin:
Manoonkitiwongsa PS, Schultz RL.
Proper nomenclature of formaldehyde and paraformaldehyde fixatives for
histochemistry. Histochem J. 2002 Jun-Jul;34(6-7):365-7.

This paper is nice-reading and its helpfull to avoid discrepancy in
describing fixatives in literature.

Subrata Biswas, MD
Dept of Nephrology
University of Campinas
SP, Brazil.




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

Message: 6
Date: Wed, 21 Jul 2004 13:58:11 -0400
From: "Flynn, Evelyn" 
Subject: RE: [Histonet] Blood vessel antibodies for rabbit models
To: degaboh@rice.edu, histonet@lists.utsouthwestern.edu
Message-ID:
	
Content-Type: text/plain; charset=iso-8859-1

Dear Zarana,
     Some years ago I purchased a polyclonal goat anti-human Factor VIII (von Willebrand)
which cross-reacted with rabbit, from Incstar Corporation.  The staining with FFPE
sections was very satisfactory.  However, I have been unable to purchase more from
this company, and I would love to find a new supplier.

Regards,
Evelyn


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu on behalf of degaboh@rice.edu
Sent: Tue 7/20/2004 5:20 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Blood vessel antibodies for rabbit models
 
 Hello all, 

I'd like to use rack the brains of all you histonetters. Can anyone
recommend antibodies to CD31 and/or CD34 that can be used in rabbit models?
Or, can anyone recommend other antibodies for blood vessels that can be used
in rabbit models?  I'm having a hard time finding these.

Thanks!

Zarana Patel
degaboh@rice.edu


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






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

Message: 7
Date: Wed, 21 Jul 2004 13:24:20 -0500
From: "Sarah Jones" 
Subject: Re: [Histonet] Oil of wintergreen
To: ,
	
Message-ID: 
Content-Type: text/plain; charset=US-ASCII

The best thing for odors is X-O odor neutralizer. 
http://www.xocorp.com/   

Sarah Jones HT(ASCP)
Dept. of Vet. Anatomy & Public Health
Histology Lab
Texas A&M University
College Station, TX 77843-4458
phone: 979-845-3177
fax:  979-458-3499


>>>  7/21/2004 7:04:55 AM >>>

Does anyone know where oil of wintergreen can be purchased?  The PA's
at our
facility used to use this product in the morgue to take care of odors
while
working on an autopsy.  I cannot locate the product and was wondering
if
anyone could help me or what else is recommended?  This is not an area
that
I am familiar with and am fairly new to my position here!!  Thanks
ahead of
time for any and all help!!

Dorothy Webb
Regions Hospital
St.Paul, Mn.

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

Message: 8
Date: Wed, 21 Jul 2004 13:38:19 -0500
From: "Sarah Jones" 
Subject: Re: [Histonet] AURANTIA
To: ,
Message-ID: 
Content-Type: text/plain; charset=ISO-8859-1

 Aurantia is a dye. Color Index number 10360.  Try looking for it's
synonym:  Imperial Yellow.  A German catalog identifies it as CI 18110
(benzol fast red B, FIAT 764).  The free amine 2,2' ,4,4'
,6,6'-hexanitrodiphenylamine appears in chemical catalogs in reagent
grade. From Conn's Biological Stains.  I have several old vials of the
dye.  I can send you one if you can't find it.  

Sarah Jones HT(ASCP)
Dept. of Vet. Anatomy & Public Health
Histology Lab
Texas A&M University
College Station, TX 77843-4458
phone: 979-845-3177
fax:  979-458-3499


>>> Jose Luis Palazon Fernandez  7/21/2004
4:59:58 AM >>>
Dear Histonet fellows

Do any of you know the comertial name of "Aurantia"? I need this
reagent but I can´t find it in the chemical product catalogs.

thanks in advance

José Luis


Universidad de Oriente-Isla Margarita-Venezuela
actualmente en: Instituto de Ciencias Marinas de Andalucia
Puerto Real, Cádiz, España.
email: jluis.palazon@icman.csic.es 

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



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

Message: 9
Date: Wed, 21 Jul 2004 19:44:00 +0100
From: "Carl" 
Subject: [Histonet] cryostat sectioning of unfixed human brains
To: 
Message-ID: <002201c46f52$b07f2320$d1039a51@home>
Content-Type: text/plain;	charset="iso-8859-1"

Be most grateful for Standard operating procedures/protocols/ info to set up system for the freezing of unfixed human brain material and cutting of cryostat sections therefrom. The tissue is not known to be infected but I appreciate that such tissue has to be treated as potentially infected.  
Thank you 


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

Message: 10
Date: Wed, 21 Jul 2004 13:47:22 -0500
From: "Charles  Scouten" 
Subject: RE: [Histonet] Agarose blocks
To: "Roberta Horner" ,
	
Message-ID:
	
	
Content-Type: text/plain;	charset="iso-8859-1"

 
What kind of microtome are you using?   Soft things need to be cut with a Vibratome.  How thin does it need to be?

Cordially,
Charles W.  Scouten, Ph.D. 
myNeuroLab.com 
5918 Evergreen Blvd. 
St. Louis, MO 63134 
Ph: 314 522 0300  
FAX  314 522 0377 
cwscouten@myneurolab.com 
http://www.myneurolab.com 


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Roberta Horner
Sent: Wednesday, July 21, 2004 12:39 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Agarose blocks

I just received tissue in agarose and I tried cutting a couple of them with very little luck.  I have never worked with this before.  Is there a trick to sectioning it?  Should it have been processed differently?  I didn't know what was in the block until I embedded it.

Thank you for any help.

Roberta Horner HT/HTL

 

Animal Diagnostic Lab

Penn State University

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



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

Message: 11
Date: Wed, 21 Jul 2004 13:51:47 -0500
From: "Charles  Scouten" 
Subject: RE: [Histonet] DECONTAMINATING CRYOSTATS
To: "Winters, Bert" ,
	
Message-ID:
	
	
Content-Type: text/plain;	charset="iso-8859-1"

I do not thing such a thing is even theoretically possible.  Kill TB germs buried in ice? 

Get a self decontaminating cryostat, and do the job overnight while you sleep.

http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=475102&catdesc=Histology+Equipment&CatThreeID=732&CatOneID=4&subcatdesc=Cryostats&idsubcategory=182




Cordially,
Charles W.  Scouten, Ph.D. 
myNeuroLab.com 
5918 Evergreen Blvd. 
St. Louis, MO 63134 
Ph: 314 522 0300  
FAX  314 522 0377 
cwscouten@myneurolab.com 
http://www.myneurolab.com 


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Winters, Bert
Sent: Wednesday, July 21, 2004 10:52 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] DECONTAMINATING CRYOSTATS


Lately we have done several frozen sections on lung cases that have turned out to be possible T.B. cases. We then have to shut down our cryostats, defrost them and decontaminate them Does anyone know of any decontamination procedures that does  not require the total defrosting of the cryostat.

                      Bert Winters, Northwest community hospital _______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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

Message: 12
Date: Wed, 21 Jul 2004 15:02:28 -0400
From: Denise Crowley 
Subject: [Histonet] Webmaster attention
To: histonet@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; charset="us-ascii" ; format="flowed"

I was trying to log in with password "digest" to change to a digest 
format and this error message came up.
thanks,
Denise


Bug in Mailman version 2.1.3

We're sorry, we hit a bug!

If you would like to help us identify the problem, please email a 
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     raise ValueError
ValueError

Python information:

Variable
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sys.version
2.2.2 (#1, Feb 24 2003, 19:13:11)  [GCC 3.2.2 20030222 (Red Hat Linux 3.2.2-4)]

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

Message: 13
Date: Wed, 21 Jul 2004 14:04:32 -0500
From: "Charles  Scouten" 
Subject: RE: [Histonet] DECONTAMINATING CRYOSTATS
To: "Winters, Bert" ,
	
Message-ID:
	
	
Content-Type: text/plain;	charset="iso-8859-1"

 
I forgot to mention, full decontamination can be done in 3.5 hours, frozen to frozen, with thaw and decontamination steps in between.  And no user interaction.

http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=475102&catdesc=Histology+Equipment&CatThreeID=732&CatOneID=4&subcatdesc=Cryostats&idsubcategory=182



Cordially,
Charles W.  Scouten, Ph.D. 
myNeuroLab.com 
5918 Evergreen Blvd. 
St. Louis, MO 63134 
Ph: 314 522 0300  
FAX  314 522 0377 
cwscouten@myneurolab.com 
http://www.myneurolab.com 


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Winters, Bert
Sent: Wednesday, July 21, 2004 10:52 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] DECONTAMINATING CRYOSTATS


Lately we have done several frozen sections on lung cases that have turned out to be possible T.B. cases. We then have to shut down our cryostats, defrost them and decontaminate them Does anyone know of any decontamination procedures that does  not require the total defrosting of the cryostat.

                      Bert Winters, Northwest community hospital _______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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

Message: 14
Date: Wed, 21 Jul 2004 14:26:25 -0500
From: "Charles  Scouten" 
Subject: RE: [Histonet] (Histonet) Sakura Auto Stainer
To: 
Message-ID:
	
	
Content-Type: text/plain;	charset="iso-8859-1"

Why not look for a stainer with 44 baths. myNeurolab.com offers the most versatile Tissue stainer in TST 44. it comes with 44 baths and will allow you to do up to 400 slides per minute. You can obtain more information about this product at the followign link. 

http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=545201&catdesc=Histology+Equipment&CatThreeID=648&CatOneID=4&subcatdesc=Tissue+Staining&idsubcategory=192

If you have any question or would like a demonstration please contact us. 


Cordially,
Charles W.  Scouten, Ph.D. 
myNeuroLab.com 
5918 Evergreen Blvd. 
St. Louis, MO 63134 
Ph: 314 522 0300  
FAX  314 522 0377 
cwscouten@myneurolab.com 
http://www.myneurolab.com 



-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jerry Wilson
Sent: Friday, July 16, 2004 11:48 PM
To: histonet@pathology.swmed.edu
Subject: [Histonet] (Histonet) Sakura Auto Stainer

I am currently considering purchasing a Sakura Automatic slide stainer- DRS 6000 (i think).  I would like to set this instrument up to stain both Histology and Cytology slides.  I understand the unit has only 27 resevoirs for reagents, but can any of the resevoirs be commonly used by both procedures?  Or, to get to the point- does anyone do this with their stainer and would they be willing to share the procedure?

Thanks
Jerry Wilson
_______________________________________________
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------------------------------

Message: 15
Date: Wed, 21 Jul 2004 15:38:00 -0400
From: james.zimmerman@pharma.novartis.com
Subject: [Histonet] C4D
To: Histonet@Pathology.swmed.edu
Message-ID:
	
	
Content-Type: text/plain; charset="us-ascii"

Hello,

Does anyone have any experience with Compliment 4 D?

If so, I would appreciate a vendor and protocol in monkey and/or  mice.


Thanks,

JPZ

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

Message: 16
Date: Wed, 21 Jul 2004 15:42:29 -0500
From: "Mitchell \(Jean\)" 
Subject: [Histonet] Darkroom Processor
To: 
Message-ID:
	
	
Content-Type: text/plain;	charset="US-ASCII"

Is anyone familiar with or currently using a MohrPro 8 Darkroom
Processor?  I am looking for some feedback on this instrument before
making the purchase.

Thanks,
Jean Mitchell
University of Wisconsin Hospital & Clinics
Department of Neurology
Madison, WI 



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

Message: 17
Date: Wed, 21 Jul 2004 15:06:09 -0500
From: "Sarah Jones" 
Subject: [Histonet] hard paraffin
To: 
Message-ID: 
Content-Type: text/plain; charset=US-ASCII

What is the hardest paraffin on the market?  I've found Gold Standard
Peel-A-Way with a melting point of 62-64 degrees C.  Does anyone have
any comments about the Gold Standard paraffins?  Years ago, a lab used
some sort of additive to their paraffin for bone that made the block
yellow in color.  Does anyone know what that could have been.  I believe
it was something like picrotin?  Ring any bells?  Thanks, Sarah

Sarah Jones HT(ASCP)
Dept. of Vet. Anatomy & Public Health
Histology Lab
Texas A&M University
College Station, TX 77843-4458
phone: 979-845-3177
fax:  979-458-3499




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

Message: 18
Date: Wed, 21 Jul 2004 17:00:33 -0400
From: "Connolly, Brett M" 
Subject: [Histonet] brightfield 3D reconstruction
To: "'HISTONET' (histonet@lists.utsouthwestern.edu)"
	
Message-ID:
	
Content-Type: text/plain

Aside from 3D reconstruction of confocal, fluorescent images, is anyone
aware of any software/hardware for doing 3D reconstruction on serial
sections of brightfield images? I have heard of VoxBlast from Vaytek, but
that's about it.

Thanks, Brett 

Brett M. Connolly, Ph.D.
Merck & Co., Inc.
MRL, Imaging Research
WP26A-3000
PO Box 4
West Point, PA 19486
PH 215-652-2501
fax. 215-652-2075
e-mail. brett_connolly@merck.com



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

Message: 19
Date: Wed, 21 Jul 2004 15:11:05 -0600
From: Gayle Callis 
Subject: [Histonet] hard paraffin additive
To: "Sarah Jones" ,
	Histonet@lists.utsouthwestern.edu
Message-ID:
	<6.0.0.22.0.20040721150518.01b1fa18@gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed

Tissue Prep 2 is hard but its melting point is not that high, and check 
with Richard Allan - I think they have one also.

I think you were thinking of piccolyte, a resin that dissolves in the 
melted paraffin.  We add this to paraffin milleniums ago, before paraffin 
technology and additives improved.  It made the paraffin  supersticky plus 
it was a total pain to deal with, a huge mess.  I recall the company that 
made it was called Hercules Corp, I am not sure you can even get it 
anymore.  It come in huge bags and big chunks in the bag.

There is a publication on this resin in J of Histotechnology back a few 
years.  Personally, you should try to find one ready to go to save 
time.  Surgipath has a blue ribbon paraffin, check with them also.

  At 02:06 PM 7/21/2004, you wrote:
>What is the hardest paraffin on the market?  I've found Gold Standard
>Peel-A-Way with a melting point of 62-64 degrees C.  Does anyone have
>any comments about the Gold Standard paraffins?  Years ago, a lab used
>some sort of additive to their paraffin for bone that made the block
>yellow in color.  Does anyone know what that could have been.  I believe
>it was something like picrotin?  Ring any bells?  Thanks, Sarah
>
>Sarah Jones HT(ASCP)
>Dept. of Vet. Anatomy & Public Health
>Histology Lab
>Texas A&M University
>College Station, TX 77843-4458
>phone: 979-845-3177
>fax:  979-458-3499
>
>
>_______________________________________________
>Histonet mailing list
>Histonet@lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)





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

Message: 20
Date: Wed, 21 Jul 2004 15:17:58 -0600
From: Gayle Callis 
Subject: Re: [Histonet] brightfield 3D reconstruction
To: "Connolly, Brett M" ,
	Histonet@lists.utsouthwestern.edu
Message-ID:
	<6.0.0.22.0.20040721151203.01b2ef90@gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed

The publication, Microscopy Today frequently has lpublications on this 
technology.  You can check out their website and do a search.  Also, people 
who do confocal and associated with core imaging facilities probably do 3D 
reconstruction are easily accessed via confocal listserver out of Buffalo, 
ask the pertinent question.   (you may already participate?)

  At 03:00 PM 7/21/2004, you wrote:
>Aside from 3D reconstruction of confocal, fluorescent images, is anyone
>aware of any software/hardware for doing 3D reconstruction on serial
>sections of brightfield images? I have heard of VoxBlast from Vaytek, but
>that's about it.
>
>Thanks, Brett

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)





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

Message: 21
Date: Wed, 21 Jul 2004 14:30:46 -0700
From: Rocan 
Subject: Re: [Histonet] Blood vessel antibodies for rabbit models
To: "'histonet@pathology.swmed.edu'" ,
	"Flynn, Evelyn" 
Message-ID: <3A27E75C-DB5D-11D8-8142-000A9589219E@mac.com>
Content-Type: text/plain;	charset=US-ASCII;	format=flowed

I think there is a goat  polyclonal against human vwf is now available 
through DAKO.  Indeed this antibody stains vessels from human and mouse 
and I would expect it to cross also with rabbit.

-----
Dr.Rocio Sierra-Honigmann
Director
Engineered Wound Repair Laboratory
Cedars Sinai Research Institute
Davis 1091
310-423-1882
Honigmannr@cshs.org	
On Jul 21, 2004, at 10:58 AM, Flynn, Evelyn wrote:

> Dear Zarana,
>      Some years ago I purchased a polyclonal goat anti-human Factor 
> VIII (von Willebrand)
> which cross-reacted with rabbit, from Incstar Corporation.  The 
> staining with FFPE
> sections was very satisfactory.  However, I have been unable to 
> purchase more from
> this company, and I would love to find a new supplier.
>
> Regards,
> Evelyn
>
>
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu on behalf of 
> degaboh@rice.edu
> Sent: Tue 7/20/2004 5:20 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Blood vessel antibodies for rabbit models
>
>  Hello all,
>
> I'd like to use rack the brains of all you histonetters. Can anyone
> recommend antibodies to CD31 and/or CD34 that can be used in rabbit 
> models?
> Or, can anyone recommend other antibodies for blood vessels that can 
> be used
> in rabbit models?  I'm having a hard time finding these.
>
> Thanks!
>
> Zarana Patel
> degaboh@rice.edu
>
>
> _______________________________________________
> 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


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

Message: 22
Date: Wed, 21 Jul 2004 14:57:04 -0700
From: "Brody,Juanita X" 
Subject: [Histonet] Indirect Immunofluorescence
To: "'histonet@pathology.swmed.edu'" 
Message-ID:
	<8220AFE580A4A34582D63C3D3B2C00EB0F60EF@cscrdemsg004.crdc.kp.org>
Content-Type: text/plain;	charset="iso-8859-1"

 We do indirect IF on skin specimens and  having problems with false positives and background staining. Is there any one doing this procedure who would be willing to share the source of money slides, control sera, and procedure?
Thanks in advance.


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

Message: 23
Date: Wed, 21 Jul 2004 20:01:11 -0400
From: 
Subject: Re: [Histonet] DECONTAMINATING CRYOSTATS
To: "Winters, Bert" ,
	
Message-ID: <008a01c46f7f$00df9940$a623d445@domainnotset.invalid>
Content-Type: text/plain;	charset="iso-8859-1"

How about not allowing frozen sections on lung for the diagnosis of
infectious diseases? TB, Pneumocystis, etc.

Do a touch prep for the fresh tissue, and stain the slide that way. Make
several slides, so can do H&E, Giemsa, GMS, Kinyoun, PASH, whatever is
needed. Fix and process the tissue used for the touch prep, for diagnosis on
a permanent section the next day.

Most of the time, the touch prep will reveal the micro-organisms. Once in a
great while, there is a false negative, which is picked up the next day on
the fixed processed block.

If it is explained to the surgeons that doing frozen sections on infectious
lung tissue puts the cryostat out of commission for the rest of the day, and
that no other surgeries, including all tumor cases, will have any FS done
for the rest of that day, they are usually willing to accept the results of
the touch preps.

Occasionally, an infectious lung will still slip through, but usually
because they thought the x-ray showed a tumor not an infection, or there is
infection with the tumor.

But there is no need to do a FS on an infectious case. Like you said, it
decommissions the cryostat, to say nothing of the potential biohazardous
risk the person doing the sectioning is in, if they accidentally cut
themselves.

You will need the support/backing of the pathologists. But they should
support you, knowing that the cryostat won't be available for the rest of
the day. (Even if they aren't worried about the risk to the sectioner.)

Peggy A. Wenk, HTL(ASCP)SLS
William Beaumont Hospital
Royal Oak, MI 48073

----- Original Message -----
From: "Winters, Bert" 
To: 
Sent: Wednesday, July 21, 2004 11:52 AM
Subject: [Histonet] DECONTAMINATING CRYOSTATS



Lately we have done several frozen sections on lung cases that have turned
out to be possible T.B. cases. We then have to shut down our cryostats,
defrost them and decontaminate them Does anyone know of any decontamination
procedures that does  not require the total defrosting of the cryostat.

                      Bert Winters, Northwest community hospital
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------------------------------

Message: 24
Date: Wed, 21 Jul 2004 18:02:15 -0700
From: Swaram 
Subject: [Histonet] Pan Melanoma cocktail from "Not" Mouse...??
To: HISTONET 
Message-ID: <40FF1217.5090905@myrealbox.com>
Content-Type: text/plain; charset="us-ascii"


   Dear Histonetters,
                                    I am in need of Pan Melanoma cocktail
   from  any  species other than Mouse. I have already tried one other Ab
   (polyclonal  Rabbit  Mart-1/Melan  A)  and  it  did  not do a good job
   compared  to  the  Pan  Melanoma cocktail (containing HMB-45, MART-1 &
   Tyrosinase).  The Ab should is to be used for IF and on paraffin fixed
   tissue.  Anybody  who  makes  them, kindly contact me. Any information
   from histonetters would be appreciated.
   Thanks,
   Swaram.


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

Message: 25
Date: Wed, 21 Jul 2004 20:57:14 -0500
From: "Sarah Jones" 
Subject: Re: [Histonet] hard paraffin additive
To: ,
Message-ID: 
Content-Type: text/plain; charset=US-ASCII

Thanks for your reply Gayle.  From the info you gave, I found piccolyte
on the Hercules web site.  Still looking for the J of H article. 
Wouldn't paraffin hardness be a factor of melting point, with the higher
melting point being the harder paraffin?   Thanks, Sarah

>>> Gayle Callis  7/21/2004 4:11:05 PM >>>
Tissue Prep 2 is hard but its melting point is not that high, and check

with Richard Allan - I think they have one also.

I think you were thinking of piccolyte, a resin that dissolves in the 
melted paraffin.  We add this to paraffin milleniums ago, before
paraffin 
technology and additives improved.  It made the paraffin  supersticky
plus 
it was a total pain to deal with, a huge mess.  I recall the company
that 
made it was called Hercules Corp, I am not sure you can even get it 
anymore.  It come in huge bags and big chunks in the bag.

There is a publication on this resin in J of Histotechnology back a few

years.  Personally, you should try to find one ready to go to save 
time.  Surgipath has a blue ribbon paraffin, check with them also.

  At 02:06 PM 7/21/2004, you wrote:
>What is the hardest paraffin on the market?  I've found Gold Standard
>Peel-A-Way with a melting point of 62-64 degrees C.  Does anyone have
>any comments about the Gold Standard paraffins?  Years ago, a lab
used
>some sort of additive to their paraffin for bone that made the block
>yellow in color.  Does anyone know what that could have been.  I
believe
>it was something like picrotin?  Ring any bells?  Thanks, Sarah
>
>Sarah Jones HT(ASCP)
>Dept. of Vet. Anatomy & Public Health
>Histology Lab
>Texas A&M University
>College Station, TX 77843-4458
>phone: 979-845-3177
>fax:  979-458-3499
>
>
>_______________________________________________
>Histonet mailing list
>Histonet@lists.utsouthwestern.edu 
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)



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



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

Message: 26
Date: Thu, 22 Jul 2004 08:54:02 +0100
From: "Brett, Lawrence" 
Subject: RE: [Histonet] Galectin-3
To: "'Sawrenko, Christina'" , "Histonet
	(E-mail)" 	
Message-ID:
	<857344E7D8F90240935A288998A89D811E774B@wgh-ex1.luht.scot.nhs.uk>
Content-Type: text/plain; charset="iso-8859-1"

Yes we use GAL 3 for Thyroid tumours too, along with Cytokeratin 19.
Papillary Ca's are positive with CK19 and mostly negative with Gal 3
while follicular ca's are mostly positive with GAL 3 and mostly negative
with CK19.
See Beesley & McLaren. Histopathology 41. 3. p236 (2002).
 
Lawrence Brett
Royal Infirmary
Edinburgh
Scotland

 
 -----Original Message-----
From: Sawrenko, Christina [mailto:csawrenk@bccancer.bc.ca]
Sent: 21 July 2004 22:07
To: Brett, Lawrence
Subject: RE: [Histonet] Galectin-3



Thanks for the info.  For what application do you use the antibody?  We are
interested in using it for thyroid neoplasms.


Thanks again, 
Chris 

-----Original Message----- 
From: Brett, Lawrence [ mailto:Lawrence.Brett@luht.scot.nhs.uk
 ] 
Sent: Wednesday, July 21, 2004 8:11 AM 
To: 'Sawrenko, Christina' 
Subject: RE: [Histonet] Galectin-3 


We use Novocastra's Galectin 3 but not with CD44v6. Gal 3 works well with 
Tris-EDTA heat antigen retrieval. We use a microwave pressure cooker 
(Biogenex) and incubate for 30 mins in primary diluted at 1:150. 
Hope this is of some value. 

Lawrence Brett 
Immunocytochemistry Lab. 
Royal Infirmary 
Edinburgh 
Scotland 

-----Original Message----- 
From: Sawrenko, Christina [ mailto:csawrenk@bccancer.bc.ca
 ] 
Sent: 16 July 2004 18:21 
To: 'histonet@lists.utsouthwestern.edu' 
Subject: [Histonet] Galectin-3 


Good morning, 
One of our pathologists has asked us to research Galectin-3 for use in 
thyroid neoplasms.  Does anyone have experience with this antibody and do 
you use it in conjuction with CD44v6 (Novocastra)? 
  
Thanks in advance for your help! 
Chris Sawrenko 
Histopathology 
BC Cancer Agency 
Vancouver BC Canada 
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------------------------------

Message: 27
Date: Thu, 22 Jul 2004 09:46:53 +0100
From: Margaret Blount 
Subject: RE: [Histonet] Agarose blocks
To: 'Roberta Horner' , histonet@lists.utsouthwestern.edu
Message-ID:
	<2A70D44ECF6F1A4390DD1D98E8BEDEF211138C@mius2.medlan.cam.ac.uk>
Content-Type: text/plain;	charset="iso-8859-1"

I have cut lots of agar blocks and used the normal tissue processing
schedules of whichever lab I worked in. I usually post fixed the agar block
in Neutral Buffered Formalin for a few hours to a day after embedding
depending upon how well fixed the sample was prior to embedding in the agar
and processing. I do think your client ought to have told you what was
embedded in the agarose else how do you know how much to trim or what the
orientation is? I have used this procedure to orientate isolated hair
follicles and flat intestine samples so that I could get sections of precise
areas, so it was essential to have all the available information about the
sample.

In difficult cases, I hand processed the agar blocks to paraffin, but this
should not be necessary if you have a process that is suited to the tissue
sample itself. I always used 2% w/v aqueous agar (Difco) and maintained it
at 55 to 60C in a waterbath to keep it molten. I made a mould suited to the
sample, e.g. a cut off syringe, pipetted a layer of agar into it and allowed
it to start gelling, I then layered on my sample so as to orientate it
correctly, then pipetted a few more drops of agar on top, taking care to
allow it to cool sufficiently to avoid "cooking" my sample. To achieve the
latter, I drew up some agar into a pastette and held it at room temperature
for a few seconds to allow it to cool, trial and error taught me how long to
do this. 

I have to say that I have used agar for both paraffin and frozen sectioning
successfully. My hand process went through 70% ethanol, 3 changes of
absolute ethanol and 1 of histoclear each step being 1 hour, then overnight
in histoclear, followed by a further histoclear of 1 hour, 2 changes of wax
in the wax oven for 1 hour then embed. This worked well for me. 

I hope this helps.

Margaret

Margaret Blount
Chief Technician
Clinical Biochemistry
University of Cambridge
Addenbrooke's Hospital
Hills Road
Cambridge
CB2 2QR 
-----Original Message-----
From: Roberta Horner [mailto:rjr6@psu.edu]
Sent: Wednesday, July 21, 2004 6:39 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Agarose blocks


I just received tissue in agarose and I tried cutting a couple of them with
very little luck.  I have never worked with this before.  Is there a trick
to sectioning it?  Should it have been processed differently?  I didn't know
what was in the block until I embedded it.

Thank you for any help.

Roberta Horner HT/HTL

 

Animal Diagnostic Lab

Penn State University

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

Message: 28
Date: Thu, 22 Jul 2004 09:49:50 +0100
From: Margaret Blount 
Subject: RE: [Histonet] brightfield 3D reconstruction
To: "'Connolly, Brett M'" , 	"'HISTONET'
	(histonet@lists.utsouthwestern.edu)"
	
Message-ID:
	<2A70D44ECF6F1A4390DD1D98E8BEDEF211138D@mius2.medlan.cam.ac.uk>
Content-Type: text/plain;	charset="iso-8859-1"

I'm not sure if I'm right but I think that Olympus' software, AnalySIS can
do this, why not contact your Olympus rep?

Margaret

Margaret Blount
Chief Technician
Clinical Biochemistry
University of Cambridge
Addenbrooke's Hospital
Hills Road
Cambridge
CB2 2QR 

-----Original Message-----
From: Connolly, Brett M [mailto:brett_connolly@merck.com]
Sent: Wednesday, July 21, 2004 10:01 PM
To: 'HISTONET' (histonet@lists.utsouthwestern.edu)
Subject: [Histonet] brightfield 3D reconstruction


Aside from 3D reconstruction of confocal, fluorescent images, is anyone
aware of any software/hardware for doing 3D reconstruction on serial
sections of brightfield images? I have heard of VoxBlast from Vaytek, but
that's about it.

Thanks, Brett 

Brett M. Connolly, Ph.D.
Merck & Co., Inc.
MRL, Imaging Research
WP26A-3000
PO Box 4
West Point, PA 19486
PH 215-652-2501
fax. 215-652-2075
e-mail. brett_connolly@merck.com



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End of Histonet Digest, Vol 8, Issue 32
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