[Histonet] Re: frozen sections falling off slides

From:"Susan Wert"



Response to problem with frozen sections falling off slides.  

We have long worked with frozen sections for IHC and ISH (with up to 21
washes!) without losing sections.  We use polysine coated slides not
superfrost or charged slides.  Let sections dry on the slides at room
temperature overnight and then fix with 2-4% PFA for 5-10 minutes.  Lay
slides flat on absorbent paper in the hood and flood with the fixative. 
Rinse in PPBS.   For more difficult tissues, we also use silanated
slides or silylated slides.  Again dry overnight at room temperature and
fix with 2-4% PFA for 5-10 minutes.  The alehydes in the tissue will thn
form covalent bonds with the silane coating on the slide.   Fixation is
required and sections must be dry.



Cordially,

Susan E. Wert, Ph.D.
Associate Professor of Pediatrics
Director, Molecular Morphology Core
Division of Pulmonary Biology, ML 7029
Cincinnati Children's Hospital Medical Center
3333 Burnet Avenue
Cincinnati, Ohio 45229-3039
TEL:		   513-636-4297 (office, voice mail)
		         513-636-3522 (laboratory)
FAX:	    513-636-7868
E-mail:	 Susan.Wert@CCHMC.org

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Today's Topics:

   1. Re: Sakura embedding station TEC 5 (Dawn Cowie)
   2. double-staining with two rabbit antibodies, Zenon-kit
      (Yves Heremans)
   3. Binding pathology reports (Stacy McLaughlin)
   4. RE: Sakura embedding station TEC 5 (Trajkovic, Dusko)
   5. negatives (Renko, Heather D.)
   6. double-staining with two rabbit antibodies, Zenon-kit 
      (AGrobe2555@aol.com)
   7. RE: Sakura embedding station TEC 5 (Lynette Pavelich)
   8. Grosslab operation instructions (Stacy McLaughlin)
   9. Binding Pathology reports-THANK YOU!! (Stacy McLaughlin)
  10. AW: [Histonet] negatives (Gudrun Lang)
  11. RE: negatives (Cazares, Ruth)
  12. frozen sections falling off slides (Gayle Callis)
  13. RE: Sakura embedding station TEC 5 (Alan Bishop)
  14. RE: Sakura embedding station TEC 5 (Dawn Cowie)
  15. RE: Sakura embedding station TEC 5
      (Bartlett, Jeanine (CDC/CCID/NCZVED))
  16. Maryland Histo/Cyto positions (Michael LaFriniere)
  17. CD61 (Mildred Fail)
  18. RE: CD61 (Sebree Linda A.)
  19. Missouri Society for Histotechnology - Annual Meeting -	May
      31, June 1-2, 2007 - The Lodge of the Ozarks, Branson, Missouri
      (Johnson, Teri)
  20. RE: delivery time of last tray of H&E (Dawn Cowie)
  21. Re: delivery time of last tray of H&E (Kathleen Roberts)


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

Message: 1
Date: Tue, 3 Apr 2007 11:05:02 -0700 (PDT)
From: Dawn Cowie 
Subject: Re: [Histonet] Sakura embedding station TEC 5
To: Amy Lee , histonet
	
Message-ID: <382174.76486.qm@web81007.mail.mud.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Amy,
  The embedding station I like to work with the best is the Leica
EG1160. We purchased a Sakura TEC5. The one quirk we noticed is that you
cannot program it to come on before midnight and go off the next
morning. For example, we start embedding at 10:30pm and wanted it to go
off at 9am the next day. If it's before midnight it won't let you
program it this way. Sakura doesn't have an answer for this.
  Otherwise, its a good instrument.
   
  Dawn

Amy Lee  wrote:
  Hello histonetters,

Is anybody using this equipment? How do you like/dislike it? We are
looking for a new embedding station. Any input is highly appreciated! 

Amy


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

Message: 2
Date: Tue,  3 Apr 2007 20:11:51 +0200 (CEST)
From: Yves Heremans 
Subject: [Histonet] double-staining with two rabbit antibodies,
	Zenon-kit
To: histonet@lists.utsouthwestern.edu
Message-ID: <20070403181151.020EA532@bonito.ulb.ac.be>

Dear All,

We need to double-stain with two antibodies made in rabbit and would
prefer to use fluorescent detection for both. I know that Molecular
Probes has Zenon kits for differential fluorescent labeling of rabbit
antibodies, allowing you to use multiple antibodies from the same
species, however these kits require that you know the concentration of
the antibodies to be labeled. Since our rabbit antibodies are not
affinity-purified but consist of whole serum, is there a way around this
and use the Zenon kit anyway ? 

Yves



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

Message: 3
Date: Tue, 3 Apr 2007 14:38:08 -0400
From: "Stacy McLaughlin" 
Subject: [Histonet] Binding pathology reports
To: 
Message-ID:
	
	
Content-Type: text/plain;	charset="us-ascii"

Hi,

I'd like to know how many of you have your pathology reports bound into
"book form".  Is there a requirement to do this?

My reason for asking- our pathology secretary insists this needs to be
done, she says all hospitals she knows do this. 

It is taking up lots of storage space.  All of the reports are in our
computer system. 

Thanks for your help!

Stacy McLaughlin



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

Message: 4
Date: Tue, 3 Apr 2007 11:44:52 -0700
From: "Trajkovic, Dusko" 
Subject: RE: [Histonet] Sakura embedding station TEC 5
To: "Amy Lee" ,	"histonet"
	
Message-ID:
	<3AD0BD3142459B4E9B12CBEAFF2B89B20431151D@lajamrexm01.amer.pfizer.com>
Content-Type: text/plain;	charset="us-ascii"

We have a Sekura TEC 5 and a Leica. Everyone (4 colleagues) prefers the
Sekura. On Sakura, paraffin, cassette, and embedding surface are on all
of the time. Whenever we need to embed, the cryo chamber is turned on
and once the work is done, cryo is turned off. Try doing that with a
Leica. Either everything is on, or everything is off. Smart engineering.
Dusko  

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Lee
Sent: Tuesday, April 03, 2007 9:35 AM
To: histonet
Subject: [Histonet] Sakura embedding station TEC 5

Hello histonetters,
   
  Is anybody using this equipment? How do you like/dislike it? We are
looking for a new embedding station. Any input is highly appreciated! 
   
  Amy

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

Message: 5
Date: Tue, 3 Apr 2007 14:04:01 -0500
From: "Renko, Heather D." 
Subject: [Histonet] negatives
To: histonet@lists.utsouthwestern.edu
Message-ID:
	<40026EDDE64CDA47AB382C52619ACD3C06612CCA@pmc-rfd-mx01.intranet.osfnet.org>
	
Content-Type: text/plain; charset=iso-8859-1

Histo netters:

I got a call from a fellow tech asking me if I run a negative control on
my special stains and my H&E slides.  I was completely stumped because I
have always used my internal negative and an a positive for each stain. 
My explanation is that the positive is for the test and the negative is
for the patient.  I have only run negative for higher testing such as
IHC, ISH, etc.  Can someone help me to help my fellow tech with the
explanation.

Nancy washkowiak, Ottawa Community Hospital
'nwash_24@yahoo.com'

Heather Renko, Histology Coordinator
OSF Saint Anthony Medical Center
5666 East State Street
Rockford, Illinois 61108
815-395-5410
Heather.D.Renko@osfhealthcare.org


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

Message: 6
Date: Tue, 3 Apr 2007 15:05:18 EDT
From: AGrobe2555@aol.com
Subject: [Histonet] double-staining with two rabbit antibodies,
	Zenon-kit 
To: histonet@lists.utsouthwestern.edu
Message-ID: 
Content-Type: text/plain; charset="US-ASCII"

Yves,
Just do the complete stain for one (block, rabbit primary#1, fluorescent
 
secondary#1), block well and then do the second (rabbit primary#2,
fluorescent  
secondary#2).  Use two different color anti-rabbit fluorophores.  I 
have done 
this in the past with no apparent problems.    
 
Another option is to pre-react the two primaries with different
anti-rabbit  
fluorophores in separate tubes, and then add to the section. 
 
Albert
 
Albert C.  Grobe, PhD
International Heart Institute of Montana Foundation
Tissue  Engineering Lab, Saint Patrick Hospital




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

Message: 7
Date: Tue, 03 Apr 2007 15:10:16 -0400
From: "Lynette Pavelich" 
Subject: RE: [Histonet] Sakura embedding station TEC 5
To: ,,
	
Message-ID: <46126E59020000EE00012E44@smtp-gw.hurleymc.com>
Content-Type: text/plain; charset=US-ASCII

Amy,
Our hospital has a Leica EG1040H (hot plate) and a EG1040C (cold plate).
 These two units are separate.  This makes it cheaper to replace one if
one or the other 'dies'.  You have the option on the heating side to
program what time you want it to start heating, what days to heat, or
the option to always have it "on".  The cold side has a "on" or "off"
switch.  We like the equipment.  They have made some very good changes
with the wax collection drawer on the heat side and we are very happy.
hope this helped,
Lynette

Lynette Pavelich, HT(ASCP)
Histology Supervisor
Hurley Medical Center
One Hurley Plaza
Flint, MI  48503

ph:  810-257-9948
fax:  810-762-7082
>>> "Trajkovic, Dusko"  04/03/07 2:44 PM >>>
We have a Sekura TEC 5 and a Leica. Everyone (4 colleagues) prefers the
Sekura. On Sakura, paraffin, cassette, and embedding surface are on all
of the time. Whenever we need to embed, the cryo chamber is turned on
and once the work is done, cryo is turned off. Try doing that with a
Leica. Either everything is on, or everything is off. Smart engineering.
Dusko  

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Lee
Sent: Tuesday, April 03, 2007 9:35 AM
To: histonet
Subject: [Histonet] Sakura embedding station TEC 5

Hello histonetters,
   
  Is anybody using this equipment? How do you like/dislike it? We are
looking for a new embedding station. Any input is highly appreciated! 
   
  Amy

 
---------------------------------
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Go to the Yahoo! Mail Q&A for great tips from Yahoo! Answers users.
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LEGAL NOTICE
Unless expressly stated otherwise, this message is confidential and may
be privileged.  It is intended for the addressee(s) only.  Access to
this E-mail by anyone else is unauthorized.  If you are not an
addressee, any disclosure or copying of the contents of this E-mail or
any action taken (or not taken) in reliance on it is unauthorized and
may be unlawful.  If you are not an addressee, please inform the sender
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------------------------------

Message: 8
Date: Tue, 3 Apr 2007 15:12:08 -0400
From: "Stacy McLaughlin" 
Subject: [Histonet] Grosslab operation instructions
To: 
Message-ID:
	
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