[Histonet] Histo gel users?

Van Eyck, Deb deb.vaneyck <@t> phci.org
Fri Apr 15 16:08:12 CDT 2011


For those of you that use Histogel to make cell blocks or with biopsy specimens - is it necessary to have your specimen completely fixed prior to making your pellet. Or does the NBF permeate the gel? 

What about using it on specimens that are going to be tested for prognostic markers? Like Her 2 neu?  Thanks Deb.

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of histonet-request <@t> lists.utsouthwestern.edu
Sent: Wednesday, April 13, 2011 9:10 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 89, Issue 13

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Today's Topics:

   1. Immunofluorescence and FISH Combined stating (Diane Kolins)
   2. Von kossa stain (Webb, Dorothy L)
   3. RE: Von kossa stain (Breeden, Sara)
   4. Re: Histonet Digest, Vol 89, Issue 12 (Mark Elliott)
   5. AW: [Histonet] Von kossa stain (Gudrun Lang)
   6. Re: AW: [Histonet] Von kossa stain
      (Grantham, Andrea L - (algranth))
   7. Staining using a Plastic Sytem (Mahesh Polavarapu)
   8. Biopsy program for Sakura VIP 5/6 Processor (Evans, Andria B)
   9. Re: Milestone MW (Rene J Buesa)
  10. Re: Von kossa stain (Rene J Buesa)
  11. RE: Von kossa stain (Jack Ratliff)
  12. RE: Counterstain for dual chromogenic (Tony Henwood)
  13. RE: Von kossa stain (Tony Henwood)
  14. RE: Biopsy program for Sakura VIP 5/6 Processor (Tony Henwood)
  15. RE: RE: Biopsy program for Sakura VIP 5/6 Processor
      (WILLIAM DESALVO)
  16. RE: RE: Biopsy program for Sakura VIP 5/6 Processor (Tony Henwood)
  17. Update regarding question on Plastic Embedding (Mahesh Polavarapu)
  18. SP3 HER2 mAb (Richard Cartun)
  19. FW: 1 H&E slide vs. 2 (Eugenia Thomas)
  20. Ventana ultra (Barbara.Crill <@t> LPNT.net)
  21. RE: FW: 1 H&E slide vs. 2 (Monfils, Paul)
  22. RE: Ventana ultra (Sebree Linda A)


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

Message: 1
Date: Tue, 12 Apr 2011 13:23:03 -0400
From: Diane Kolins <Diane <@t> bioview.co.il>
Subject: [Histonet] Immunofluorescence and FISH Combined stating
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<!&!AAAAAAAAAAAYAAAAAAAAAKxbzBNp1TJAsnIO9XHEfgnCgAAAEAAAAKFMd/+8AfZLjHzeYTbktzQBAAAAAA==@bioview.co.il>
	
Content-Type: text/plain; charset=us-ascii

Where can I find a procedure for dual staining of blood and bone marrow
smears using Vector anti-kappa and anti-lambda tagged with Coumarine
followed by FISH hybridization. I am able to see what I think are plasma
cells by their fluorescent blue cytoplasm and on the same slide (changing
filters) I can see the red and green signal patterns.

 

From: :  diane <@t> bioview.co.il

 

 



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

Message: 2
Date: Tue, 12 Apr 2011 12:35:18 -0500
From: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>
Subject: [Histonet] Von kossa stain
To: "'histonet <@t> lists.utsouthwestern.edu'"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<65365F35C0F2EF4D846EC3CA73E49C43010F851054C1 <@t> HPEMX3.HealthPartners.int>
	
Content-Type: text/plain; charset="us-ascii"

What is everyone using for their "light" when developing the silver in the VonKossa stain when you have no sunlight to use?  We used to use a 60 watt lamp, but haven't done one for years and am bringing this stain back to our repetiore due to pathologist request.  Thanks much!

Dorothy Webb, HT (ASCP)
Regions Histology Technical Supervisor
651-254-2962




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

Message: 3
Date: Tue, 12 Apr 2011 11:52:49 -0600
From: "Breeden, Sara" <sbreeden <@t> nmda.nmsu.edu>
Subject: RE: [Histonet] Von kossa stain
To: "Webb, Dorothy L" <Dorothy.L.Webb <@t> HealthPartners.Com>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<4D14F0FC9316DD41972D5F03C070908B02E477BD <@t> nmdamailsvr.nmda.ad.nmsu.edu>
	
Content-Type: text/plain;	charset="us-ascii"

Funny you should ask... just last week I had two requests for a Von
Kossa.  That's when I found out that, despite the fact that we just
moved into a brand new building with all the bells and whistles, there
was not one single UV light in any of the many hoods that had been
installed.  Heaven forbid  I could even find an incandescent bulb (we
are a "green building").  So, I prepped my slides, put them in a clear
glass Coplin jar and parked the jar on the hood of my car for an hour.

Works like a charm.  This all depends, naturally, on (1) if you even
HAVE sunshine where you live; (2) how far a walk it is to the sunshine,
and (3) whether you have a car hood on which to park the Coplin jar.
Needless to say, out of the line of sight of curious onlookers with
sticky fingers and no business wondering what that glass jar is...
Nonetheless, it works just fine!  May the Force be with you.



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

Message: 4
Date: Tue, 12 Apr 2011 11:16:16 -0700
From: "Mark Elliott" <Mark.Elliott <@t> hli.ubc.ca>
Subject: [Histonet] Re: Histonet Digest, Vol 89, Issue 12
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <4DA43480020000D60006111F <@t> mail.mrl.ubc.ca>
Content-Type: text/plain; charset=US-ASCII


Date: Tue, 12 Apr 2011 11:37:17 -0400
From: Eva Permaul <eca9 <@t> georgetown.edu>
Subject: [Histonet] Counterstain for dual chromogenic
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <4DA471AD.3040002 <@t> georgetown.edu>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

Eva
 
I have been doing some double labelling using Vulcan Fast Red from Biocare and  Vector Blue Alkaline Phosphatase Substrate Kit III from Vector Labs.  For counter stain I use Vector Methyl Green and the combination works great and looks quite nice.
 
Mark
 
Good morning,
I have been attempting to do some dual chromogenic staining. I am using 
a mouse and a rabbit antibody. For the mouse antibody I am using an 
HRP-mouse secondary followed by AEC for visualization. For the rabbit 
antibody I am using a biotinylated rabbit secondary, followed by ABC-AP 
and vector blue. My problem is what to use as a counterstain. From 
everything I have read so far there isn't one that would work without 
having some problems. I was thinking of trying a very light Hematoxylin 
and see if it doesn't disrupt the vector blue visualization too much. 
Does anyone else have any suggestions?
Thanks,
Eva

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

Message: 5
Date: Tue, 12 Apr 2011 20:22:22 +0200
From: "Gudrun Lang" <gu.lang <@t> gmx.at>
Subject: AW: [Histonet] Von kossa stain
To: "'Webb, Dorothy L'" <Dorothy.L.Webb <@t> HealthPartners.Com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <13B705C625E74F1BB9337593D57C3658 <@t> dielangs.at>
Content-Type: text/plain;	charset="iso-8859-1"

WE use just a simple desk-lamp and put the coplin jar directly under the
bulb. And the desk-lamp is placed in the window.

Bye Gudrun

-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Webb,
Dorothy L
Gesendet: Dienstag, 12. April 2011 19:35
An: 'histonet <@t> lists.utsouthwestern.edu'
Betreff: [Histonet] Von kossa stain

What is everyone using for their "light" when developing the silver in the
VonKossa stain when you have no sunlight to use?  We used to use a 60 watt
lamp, but haven't done one for years and am bringing this stain back to our
repetiore due to pathologist request.  Thanks much!

Dorothy Webb, HT (ASCP)
Regions Histology Technical Supervisor
651-254-2962




  ________________________________
This e-mail and any files transmitted with it are confidential and are
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------------------------------

Message: 6
Date: Tue, 12 Apr 2011 11:35:47 -0700
From: "Grantham, Andrea L - (algranth)" <algranth <@t> email.arizona.edu>
Subject: Re: AW: [Histonet] Von kossa stain
Cc: HISTONET <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <59222C2B-42C1-4EB6-8233-ED0FB7FCA354 <@t> email.arizona.edu>
Content-Type: text/plain; charset="iso-8859-1"

We have intense sun here but my windows face the wrong way! At least I have windows. 
I just use a light bulb from the lamp we have over the coverslipping station. Works great.

Andi



On Apr 12, 2011, at 11:22 AM, Gudrun Lang wrote:

> WE use just a simple desk-lamp and put the coplin jar directly under the
> bulb. And the desk-lamp is placed in the window.
> 
> Bye Gudrun
> 
> -----Ursprüngliche Nachricht-----
> Von: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Webb,
> Dorothy L
> Gesendet: Dienstag, 12. April 2011 19:35
> An: 'histonet <@t> lists.utsouthwestern.edu'
> Betreff: [Histonet] Von kossa stain
> 
> What is everyone using for their "light" when developing the silver in the
> VonKossa stain when you have no sunlight to use?  We used to use a 60 watt
> lamp, but haven't done one for years and am bringing this stain back to our
> repetiore due to pathologist request.  Thanks much!
> 
> Dorothy Webb, HT (ASCP)
> Regions Histology Technical Supervisor
> 651-254-2962
> 
> 
> 
> 
>  ________________________________
> This e-mail and any files transmitted with it are confidential and are
> intended solely for the use of the individual or entity to whom they are
> addressed. If you are not the intended recipient or the individual
> responsible for delivering the e-mail to the intended recipient, please be
> advised that you have received this e-mail in error and that any use,
> dissemination, forwarding, printing, or copying of this e-mail is strictly
> prohibited.
> 
> If you have received this e-mail in error, please immediately notify the
> HealthPartners Support Center by telephone at (952) 967-6600. You will be
> reimbursed for reasonable costs incurred in notifying us. HealthPartners
> R001.0
> _______________________________________________
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> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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> 
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> 





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

Message: 7
Date: Tue, 12 Apr 2011 13:56:53 -0500
From: Mahesh Polavarapu <polavarapu.mahesh <@t> gmail.com>
Subject: [Histonet] Staining using a Plastic Sytem
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <BANLkTikXBMVMdTC70GXe4_o3_N=ujYyOWw <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Hi,

Does anyone have experience embedding and staining in plastic? We have
rabbit rotator cuff tendons that are being embedded in plastic. I am trying
to figure out what the process is and which stains to use in order to
visualize (and quantify) blood vessels and collagen deposition at the Bone
(Humeral Head) - Tendon (Infraspinatus) interface.

Thanks,
Mahesh


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

Message: 8
Date: Tue, 12 Apr 2011 14:57:18 -0400
From: "Evans, Andria B" <aevans3 <@t> lghealth.org>
Subject: [Histonet] Biopsy program for Sakura VIP 5/6 Processor
To: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<4182FDF23D7C9948BC41C4C082C3A54F021557B3AA60 <@t> MAIL-AG-CLUSTER.lha.org>
Content-Type: text/plain;	charset="iso-8859-1"

Our lab is currently looking for a way to shorten our Biopsy processing program without compromising the patient specimen.  We do have an issue with our GI's being very dry, which causes us to have to soak between each level taken and also causes a lot of chatter.  Also we have a goal to do a run during the day to improve turn around time.   Here is what our current protocol is....

Formalin    1 min       37degrees
Formalin    15 mins    37degrees
70             14 mins    40 degrees
95             14 mins    40 degrees
95             9 mins      40 degrees
100           9 mins      40 degrees
100           7 mins      40 degrees
100           4 mins      40 degrees
Xylene       23 mins    no heat
Xylene       15 mins    no heat
Paraffin      20 mins   60 degrees
Paraffin      18 mins   60 degrees
Paraffin      10 mins   60 degrees
Paraffin      0 mins     60 degrees

All the steps are set on a fast mix setting.  All of our biopsy specimens are put into sponges.

Any feedback would be greatly appreciated.

Andria B Evans HTL(ASCP)CM
Lancaster General Hospital
555 North Duke Street
Lancaster, PA  17604
(717)544-5511 ext: 77329
aevans3 <@t> lgheath.org<mailto:aevans3 <@t> lgheath.org>
This email was sent securely from the LGHealth Email Service

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Message: 9
Date: Tue, 12 Apr 2011 12:27:49 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Milestone MW
To: Histonet <@t> lists.utsouthwestern.edu, SHANE NELSON
	<nelsonrnch <@t> verizon.net>
Message-ID: <809117.78546.qm <@t> web65715.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Model "KOS" by Milestone is capable of tissue processing, decalcification, special stains, fixation, gross hardening, and antigen retrieval.
René J.

--- On Tue, 4/12/11, SHANE NELSON <nelsonrnch <@t> verizon.net> wrote:


From: SHANE NELSON <nelsonrnch <@t> verizon.net>
Subject: [Histonet] Milestone MW
To: Histonet <@t> lists.utsouthwestern.edu
Date: Tuesday, April 12, 2011, 12:30 PM


Margret,
Which microwave would that be
 
THANK YOU,
 
PATTI RUBEN-NELSON  H.T.(ASCP) 
LABORATORY CONSULTANT
SUPERVISOR/DESERT GASTROENTEROLOGY CONSULTANTS
35-900 Bob Hope Drive
Suite 275
Rancho Mirage, Ca.  92270
cell (909) 841-9761 / wk (760) 321-2500
nelsonrnch <@t> verizon.net
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------------------------------

Message: 10
Date: Tue, 12 Apr 2011 12:30:26 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Von kossa stain
To: "'histonet <@t> lists.utsouthwestern.edu'"
	<histonet <@t> lists.utsouthwestern.edu>, 	Dorothy LWebb
	<Dorothy.L.Webb <@t> HealthPartners.Com>
Message-ID: <145774.98333.qm <@t> web65704.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

You can use any strong intensity microscope light. The stronger the intensity of the light the better, but the reduction has a cumulative effect so even not very strong intensity bulbs can be effective.
René J.

--- On Tue, 4/12/11, Webb, Dorothy L <Dorothy.L.Webb <@t> HealthPartners.Com> wrote:


From: Webb, Dorothy L <Dorothy.L.Webb <@t> HealthPartners.Com>
Subject: [Histonet] Von kossa stain
To: "'histonet <@t> lists.utsouthwestern.edu'" <histonet <@t> lists.utsouthwestern.edu>
Date: Tuesday, April 12, 2011, 1:35 PM


What is everyone using for their "light" when developing the silver in the VonKossa stain when you have no sunlight to use?  We used to use a 60 watt lamp, but haven't done one for years and am bringing this stain back to our repetiore due to pathologist request.  Thanks much!

Dorothy Webb, HT (ASCP)
Regions Histology Technical Supervisor
651-254-2962




  ________________________________
This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited.

If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0
_______________________________________________
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------------------------------

Message: 11
Date: Tue, 12 Apr 2011 15:41:54 -0400
From: Jack Ratliff <ratliffjack <@t> hotmail.com>
Subject: RE: [Histonet] Von kossa stain
To: <dorothy.l.webb <@t> healthpartners.com>, Histonet
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <BLU167-w542F31C988F23C9B15EFF9AEAB0 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"


I purchase a kit from Dorn and Hart Microedge and use sodium carbonate-formaldehyde solution to chemically develop.

Stain in silver nitrate for 5 min in the dark, 3 fresh DI rinses for 1 min each in the dark, then sodium carbonate-formaldehyde solution for 2 min in the dark, 2 fresh DI rinses (normal light) for 1 min each, 30 seconds in Farmer's Diminisher (Sodium thiosulfate-potassium ferricyanide soluiton to stop reaction) and a running tap water rinse for 10 min.
 
Jack
 

 
> From: Dorothy.L.Webb <@t> HealthPartners.Com
> To: histonet <@t> lists.utsouthwestern.edu
> Date: Tue, 12 Apr 2011 12:35:18 -0500
> Subject: [Histonet] Von kossa stain
> 
> What is everyone using for their "light" when developing the silver in the VonKossa stain when you have no sunlight to use? We used to use a 60 watt lamp, but haven't done one for years and am bringing this stain back to our repetiore due to pathologist request. Thanks much!
> 
> Dorothy Webb, HT (ASCP)
> Regions Histology Technical Supervisor
> 651-254-2962
> 
> 
> 
> 
> ________________________________
> This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited.
> 
> If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. HealthPartners R001.0
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 		 	   		  

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

Message: 12
Date: Tue, 12 Apr 2011 23:11:35 +0000
From: Tony Henwood <AnthonyH <@t> chw.edu.au>
Subject: RE: [Histonet] Counterstain for dual chromogenic
To: "'Eva Permaul'" <eca9 <@t> georgetown.edu>,
	"histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <6D6BD1DE8A5571489398B392A38A71571884E925 <@t> xmdb02.nch.kids>
Content-Type: text/plain; charset="us-ascii"

Try ethyl green:
1.	0.1N Acetic Acid
	Add 6ml of glacial acetic acid to 1000 ml of distilled water.  
2.	0.1N Sodium Acetate
Add 4.102 g of sodium acetate to 500 ml of distilled water
3.	pH 4.2 buffer
Add 755 ml of 0.1N acetic acid to 264 ml of 0.1N Sodium Acetate and adjust the pH to 4.2 with 1M NaOH
4.	Ethyl Green Solution
To 100ml of buffer add 2g Ethyl Green (CI 42590)
Can be stored at room temp.  
If staining turns bluish than the solution has started to deteriorate.

Stain for 5 minutes. Do not rinse in water for too long, it tends to extract the green staining

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Eva Permaul
Sent: Wednesday, 13 April 2011 1:37 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Counterstain for dual chromogenic

Good morning,
I have been attempting to do some dual chromogenic staining. I am using a mouse and a rabbit antibody. For the mouse antibody I am using an HRP-mouse secondary followed by AEC for visualization. For the rabbit antibody I am using a biotinylated rabbit secondary, followed by ABC-AP and vector blue. My problem is what to use as a counterstain. From everything I have read so far there isn't one that would work without having some problems. I was thinking of trying a very light Hematoxylin and see if it doesn't disrupt the vector blue visualization too much. 
Does anyone else have any suggestions?
Thanks,
Eva

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

Message: 13
Date: Tue, 12 Apr 2011 23:22:21 +0000
From: Tony Henwood <AnthonyH <@t> chw.edu.au>
Subject: RE: [Histonet] Von kossa stain
To: "'Breeden, Sara'" <sbreeden <@t> nmda.nmsu.edu>, "Webb, Dorothy L"
	<Dorothy.L.Webb <@t> HealthPartners.Com>,
	"histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <6D6BD1DE8A5571489398B392A38A71571884E95A <@t> xmdb02.nch.kids>
Content-Type: text/plain; charset="us-ascii"

Interesting,

Meloan & Puchtler (1985) have drawn our attention to Von Kossa's original work on this technique. He regarded only the yellow colouration of calcium deposits during early stages of the reaction as diagnostic for calcium phosphate and credited the blackening to organic matter. Further studies showed that bright light only causes the irreversible blackening of organic matter that masks the yellow silver phosphate. When the reaction is performed in subdued light, yellow to yellowish brown silver phosphate is visualised selectively. Silver carbonate dissolves in sodium thiosulphate and cannot be demonstrated with von Kossa's technique (Meloan & Puchtler 1985 J Histotechnol 8(1):11-13.).

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Breeden, Sara
Sent: Wednesday, 13 April 2011 3:53 AM
To: Webb, Dorothy L; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Von kossa stain

Funny you should ask... just last week I had two requests for a Von Kossa.  That's when I found out that, despite the fact that we just moved into a brand new building with all the bells and whistles, there was not one single UV light in any of the many hoods that had been installed.  Heaven forbid  I could even find an incandescent bulb (we are a "green building").  So, I prepped my slides, put them in a clear glass Coplin jar and parked the jar on the hood of my car for an hour.

Works like a charm.  This all depends, naturally, on (1) if you even HAVE sunshine where you live; (2) how far a walk it is to the sunshine, and (3) whether you have a car hood on which to park the Coplin jar.
Needless to say, out of the line of sight of curious onlookers with sticky fingers and no business wondering what that glass jar is...
Nonetheless, it works just fine!  May the Force be with you.

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

*********************************************************************************
This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender.

Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead

This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses.
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------------------------------

Message: 14
Date: Tue, 12 Apr 2011 23:30:30 +0000
From: Tony Henwood <AnthonyH <@t> chw.edu.au>
Subject: [Histonet] RE: Biopsy program for Sakura VIP 5/6 Processor
To: "'Evans, Andria B'" <aevans3 <@t> lghealth.org>,
	"histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <6D6BD1DE8A5571489398B392A38A71571884E981 <@t> xmdb02.nch.kids>
Content-Type: text/plain; charset="us-ascii"

Increase the fixation time and make sure the air is out of the sponges (will stop formalin from getting to the tissue- a quick dunk of the cassette (with sponge and tissue) in alcohol and back into formalin will do the trick).

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Evans, Andria B
Sent: Wednesday, 13 April 2011 4:57 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Biopsy program for Sakura VIP 5/6 Processor

Our lab is currently looking for a way to shorten our Biopsy processing program without compromising the patient specimen.  We do have an issue with our GI's being very dry, which causes us to have to soak between each level taken and also causes a lot of chatter.  Also we have a goal to do a run during the day to improve turn around time.   Here is what our current protocol is....

Formalin    1 min       37degrees
Formalin    15 mins    37degrees
70             14 mins    40 degrees
95             14 mins    40 degrees
95             9 mins      40 degrees
100           9 mins      40 degrees
100           7 mins      40 degrees
100           4 mins      40 degrees
Xylene       23 mins    no heat
Xylene       15 mins    no heat
Paraffin      20 mins   60 degrees
Paraffin      18 mins   60 degrees
Paraffin      10 mins   60 degrees
Paraffin      0 mins     60 degrees

All the steps are set on a fast mix setting.  All of our biopsy specimens are put into sponges.

Any feedback would be greatly appreciated.

Andria B Evans HTL(ASCP)CM
Lancaster General Hospital
555 North Duke Street
Lancaster, PA  17604
(717)544-5511 ext: 77329
aevans3 <@t> lgheath.org<mailto:aevans3 <@t> lgheath.org>
This email was sent securely from the LGHealth Email Service

Confidentiality Notice: 
This e-mail message, including any attachments, is for the sole use of 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 e-mail and destroy all copies of the original message.

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

*********************************************************************************
This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender.

Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead

This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses.
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------------------------------

Message: 15
Date: Tue, 12 Apr 2011 17:58:36 -0600
From: WILLIAM DESALVO <wdesalvo.cac <@t> hotmail.com>
Subject: RE: [Histonet] RE: Biopsy program for Sakura VIP 5/6
	Processor
To: <anthonyh <@t> chw.edu.au>, <aevans3 <@t> lghealth.org>, histonet
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <BAY151-w119D7602D3EDDB2E17742F91AB0 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"


I would consider replacing the sponges. Tony is correct that you need to make sure the air is removed from the sponge to facilitate exchange, but with shortened processing times you will undoubtedly cause carry over from solution to solution. Yuo may want to have a small container of alcoholic formalin with sponges sitting on the gross table.

Try using a nylon tissue bag to funnel filter your small biopsies. all your biopsy sample sizes can be placed into the bag and very tiny samples are retained. Last suggestion is to consider a folded filter paper method (we name it origami, I can provide directly if you want) that uses a quick four fold process w/ forceps at the gross bench to create a pocket for the tiny samples, has a single layer between the tissue/solution and is easy to open at embedding (without "popping" of tissue samples). Avoind the unnecessary carryover of the sponge if you can.

William DeSalvo, B.S., HTL(ASCP)





> From: AnthonyH <@t> chw.edu.au
> To: aevans3 <@t> lghealth.org; histonet <@t> lists.utsouthwestern.edu
> Date: Tue, 12 Apr 2011 23:30:30 +0000
> CC: 
> Subject: [Histonet] RE: Biopsy program for Sakura VIP 5/6 Processor
> 
> Increase the fixation time and make sure the air is out of the sponges (will stop formalin from getting to the tissue- a quick dunk of the cassette (with sponge and tissue) in alcohol and back into formalin will do the trick).
> 
> Regards 
> Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
> Laboratory Manager & Senior Scientist 
> Tel: 612 9845 3306 
> Fax: 612 9845 3318 
> the children's hospital at westmead
> Cnr Hawkesbury Road and Hainsworth Street, Westmead
> Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 
> 
> 
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Evans, Andria B
> Sent: Wednesday, 13 April 2011 4:57 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Biopsy program for Sakura VIP 5/6 Processor
> 
> Our lab is currently looking for a way to shorten our Biopsy processing program without compromising the patient specimen.  We do have an issue with our GI's being very dry, which causes us to have to soak between each level taken and also causes a lot of chatter.  Also we have a goal to do a run during the day to improve turn around time.   Here is what our current protocol is....
> 
> Formalin    1 min       37degrees
> Formalin    15 mins    37degrees
> 70             14 mins    40 degrees
> 95             14 mins    40 degrees
> 95             9 mins      40 degrees
> 100           9 mins      40 degrees
> 100           7 mins      40 degrees
> 100           4 mins      40 degrees
> Xylene       23 mins    no heat
> Xylene       15 mins    no heat
> Paraffin      20 mins   60 degrees
> Paraffin      18 mins   60 degrees
> Paraffin      10 mins   60 degrees
> Paraffin      0 mins     60 degrees
> 
> All the steps are set on a fast mix setting.  All of our biopsy specimens are put into sponges.
> 
> Any feedback would be greatly appreciated.
> 
> Andria B Evans HTL(ASCP)CM
> Lancaster General Hospital
> 555 North Duke Street
> Lancaster, PA  17604
> (717)544-5511 ext: 77329
> aevans3 <@t> lgheath.org<mailto:aevans3 <@t> lgheath.org>
> This email was sent securely from the LGHealth Email Service
> 
> Confidentiality Notice: 
> This e-mail message, including any attachments, is for the sole use of 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 e-mail and destroy all copies of the original message.
> 
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> *********************************************************************************
> This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender.
> 
> Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead
> 
> This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses.

> *********************************************************************************
> 
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
 		 	   		  

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

Message: 16
Date: Wed, 13 Apr 2011 00:10:04 +0000
From: Tony Henwood <AnthonyH <@t> chw.edu.au>
Subject: RE: [Histonet] RE: Biopsy program for Sakura VIP 5/6
	Processor
To: "'WILLIAM DESALVO'" <wdesalvo.cac <@t> hotmail.com>,
	"aevans3 <@t> lghealth.org"	<aevans3 <@t> lghealth.org>, histonet
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <6D6BD1DE8A5571489398B392A38A71571884E9D3 <@t> xmdb02.nch.kids>
Content-Type: text/plain; charset="us-ascii"

I agree

Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

From: WILLIAM DESALVO [mailto:wdesalvo.cac <@t> hotmail.com]
Sent: Wednesday, 13 April 2011 9:59 AM
To: Tony Henwood; aevans3 <@t> lghealth.org; histonet
Subject: RE: [Histonet] RE: Biopsy program for Sakura VIP 5/6 Processor

I would consider replacing the sponges. Tony is correct that you need to make sure the air is removed from the sponge to facilitate exchange, but with shortened processing times you will undoubtedly cause carry over from solution to solution. Yuo may want to have a small container of alcoholic formalin with sponges sitting on the gross table.

Try using a nylon tissue bag to funnel filter your small biopsies. all your biopsy sample sizes can be placed into the bag and very tiny samples are retained. Last suggestion is to consider a folded filter paper method (we name it origami, I can provide directly if you want) that uses a quick four fold process w/ forceps at the gross bench to create a pocket for the tiny samples, has a single layer between the tissue/solution and is easy to open at embedding (without "popping" of tissue samples). Avoind the unnecessary carryover of the sponge if you can.

William DeSalvo, B.S., HTL(ASCP)





> From: AnthonyH <@t> chw.edu.au<mailto:AnthonyH <@t> chw.edu.au>
> To: aevans3 <@t> lghealth.org<mailto:aevans3 <@t> lghealth.org>; histonet <@t> lists.utsouthwestern.edu<mailto:histonet <@t> lists.utsouthwestern.edu>
> Date: Tue, 12 Apr 2011 23:30:30 +0000
> CC:
> Subject: [Histonet] RE: Biopsy program for Sakura VIP 5/6 Processor
>
> Increase the fixation time and make sure the air is out of the sponges (will stop formalin from getting to the tissue- a quick dunk of the cassette (with sponge and tissue) in alcohol and back into formalin will do the trick).
>
> Regards
> Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
> Laboratory Manager & Senior Scientist
> Tel: 612 9845 3306
> Fax: 612 9845 3318
> the children's hospital at westmead
> Cnr Hawkesbury Road and Hainsworth Street, Westmead
> Locked Bag 4001, Westmead NSW 2145, AUSTRALIA
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu<mailto:histonet-bounces <@t> lists.utsouthwestern.edu> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu]<mailto:[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]> On Behalf Of Evans, Andria B
> Sent: Wednesday, 13 April 2011 4:57 AM
> To: histonet <@t> lists.utsouthwestern.edu<mailto:histonet <@t> lists.utsouthwestern.edu>
> Subject: [Histonet] Biopsy program for Sakura VIP 5/6 Processor
>
> Our lab is currently looking for a way to shorten our Biopsy processing program without compromising the patient specimen. We do have an issue with our GI's being very dry, which causes us to have to soak between each level taken and also causes a lot of chatter. Also we have a goal to do a run during the day to improve turn around time. Here is what our current protocol is....
>
> Formalin 1 min 37degrees
> Formalin 15 mins 37degrees
> 70 14 mins 40 degrees
> 95 14 mins 40 degrees
> 95 9 mins 40 degrees
> 100 9 mins 40 degrees
> 100 7 mins 40 degrees
> 100 4 mins 40 degrees
> Xylene 23 mins no heat
> Xylene 15 mins no heat
> Paraffin 20 mins 60 degrees
> Paraffin 18 mins 60 degrees
> Paraffin 10 mins 60 degrees
> Paraffin 0 mins 60 degrees
>
> All the steps are set on a fast mix setting. All of our biopsy specimens are put into sponges.
>
> Any feedback would be greatly appreciated.
>
> Andria B Evans HTL(ASCP)CM
> Lancaster General Hospital
> 555 North Duke Street
> Lancaster, PA 17604
> (717)544-5511 ext: 77329
> aevans3 <@t> lgheath.org<mailto:aevans3 <@t> lgheath.org<mailto:aevans3 <@t> lgheath.org%3cmailto:aevans3 <@t> lgheath.org>>
> This email was sent securely from the LGHealth Email Service
>
> Confidentiality Notice:
> This e-mail message, including any attachments, is for the sole use of 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 e-mail and destroy all copies of the original message.
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu<mailto:Histonet <@t> lists.utsouthwestern.edu>
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> *********************************************************************************
> This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender.
>
> Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead
>
> This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses.

> *********************************************************************************
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu<mailto:Histonet <@t> lists.utsouthwestern.edu>
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet


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

Message: 17
Date: Tue, 12 Apr 2011 19:25:30 -0500
From: Mahesh Polavarapu <polavarapu.mahesh <@t> gmail.com>
Subject: [Histonet] Update regarding question on Plastic Embedding
To: histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <BANLkTikk_-oe3rO1KbxXd5x4Jj0kiM6gAw <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Looking for a protocol to visualize vascularization and collagen deposition
at the bone-tendon interface of a rabbit rotator cuff embedded in a plastic
system. Sections will be rather thick (~50um) b/c they are being made
through a titanium anchor. Using MMA with a cold-curing resin, Technovit
9100. Thanks in advance!

- Mahesh


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

Message: 18
Date: Tue, 12 Apr 2011 20:34:51 -0400
From: "Richard Cartun" <Rcartun <@t> harthosp.org>
Subject: [Histonet] SP3 HER2 mAb
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <4DA4B76A.7400.0077.1 <@t> harthosp.org>
Content-Type: text/plain; charset=US-ASCII

Anyone using the HER2 rabbit monoclonal antibody (clone SP3) ever see reactivity with skeletal muscle?  Thank you.

Richard

Richard W. Cartun, MS, PhD
Director, Histology & Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596 Office
(860) 545-2204 Fax





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

Message: 19
Date: Tue, 12 Apr 2011 23:14:30 -0400
From: Eugenia Thomas <eugenia902d1 <@t> hotmail.com>
Subject: [Histonet] FW: 1 H&E slide vs. 2
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <COL103-W59F151894F9360078648E8D5AA0 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"



 


From: eugenia902d1 <@t> hotmail.com
To: histonet <@t> lists.utsouthwestern.edu
Subject: 1 H&E slide vs. 2
Date: Mon, 11 Apr 2011 13:06:59 -0400




Good afternoon everyone,
 
Does anyone know of an article or statistics discussing the impact (medical diagnosing) of cutting 2 H&E slides per block verses 1 for all routine work?
 
 
Genia
 		 	   		  

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

Message: 20
Date: Wed, 13 Apr 2011 08:39:22 -0500
From: <Barbara.Crill <@t> LPNT.net>
Subject: [Histonet] Ventana ultra
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<7DA79EBDBD92BF408EF392413737878D3936B06565 <@t> NADCWPMSGCMS01.hca.corpad.net>
	
Content-Type: text/plain; charset="us-ascii"

Does anyone use the Benchmark Ultra - care to share your comments?

We are considering switching from the Benchmark XT to the Ultra but would like to hear from users about this instrument.
Is there an increase in reagent cost?

Can you really add more slides without adding time to the run?


ANTOINETTE CRILL, MBA,CT(ASCP)
TEAM LEADER ANATOMIC PATHOLOGY
DANVILLE REGIONAL MEDICAL CENTER
(O) 434.799.4470
(F) 434.773.6806
E-mail:  barbara.crill <@t> LPNT.net<mailto:barbara.crill <@t> LPNT.net>



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

Message: 21
Date: Wed, 13 Apr 2011 10:00:09 -0400
From: "Monfils, Paul" <PMonfils <@t> Lifespan.org>
Subject: RE: [Histonet] FW: 1 H&E slide vs. 2
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<4EBFF65383B74D49995298C4976D1D5E08020E55 <@t> LSRIEXCH1.lsmaster.lifespan.org>
	
Content-Type: text/plain;	charset="us-ascii"

When I was in clinical histology (I'm in research now) the pathologists
didn't even want us putting two sections on a slide, because even though
they knew a second serial section wasn't going to show anything the
first section didn't show, they felt ethically/legally bound to examine
both sections if they were there, so they were investing twice the time
and effort for the same return.

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Eugenia
Thomas
Sent: Tuesday, April 12, 2011 11:15 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] FW: 1 H&E slide vs. 2



 


From: eugenia902d1 <@t> hotmail.com
To: histonet <@t> lists.utsouthwestern.edu
Subject: 1 H&E slide vs. 2
Date: Mon, 11 Apr 2011 13:06:59 -0400




Good afternoon everyone,
 
Does anyone know of an article or statistics discussing the impact
(medical diagnosing) of cutting 2 H&E slides per block verses 1 for all
routine work?
 
 
Genia
 
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



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

Message: 22
Date: Wed, 13 Apr 2011 09:09:22 -0500
From: "Sebree Linda A" <LSebree <@t> uwhealth.org>
Subject: RE: [Histonet] Ventana ultra
To: <Barbara.Crill <@t> LPNT.net>,	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<F0B9FD4B28C80E43996FFD4408B198AE074E6D <@t> UWHC-MAIL3.uwhis.hosp.wisc.edu>
	
Content-Type: text/plain;	charset="us-ascii"

Please "Reply All" as I'd like to hear feedback also.

Thanks. 


Linda A. Sebree
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
DB1-223 VAH
600 Highland Ave.
Madison, WI 53792
(608)265-6596


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Barbara.Crill <@t> LPNT.net
Sent: Wednesday, April 13, 2011 8:39 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Ventana ultra

Does anyone use the Benchmark Ultra - care to share your comments?

We are considering switching from the Benchmark XT to the Ultra but
would like to hear from users about this instrument.
Is there an increase in reagent cost?

Can you really add more slides without adding time to the run?


ANTOINETTE CRILL, MBA,CT(ASCP)
TEAM LEADER ANATOMIC PATHOLOGY
DANVILLE REGIONAL MEDICAL CENTER
(O) 434.799.4470
(F) 434.773.6806
E-mail:  barbara.crill <@t> LPNT.net<mailto:barbara.crill <@t> LPNT.net>

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



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

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

End of Histonet Digest, Vol 89, Issue 13
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