[Histonet] RE: Histonet Digest, Vol 72, Issue 26

[Suresch, Donna L.]

RE:  Cryojane Tape Transfer System
I have found that you need to have your tape at the same temp as your
tissue to have your tissue transfer completely from the tape.  I set
everything up in the morning putting the tape in the cryostat and get
everything equilibrated first for about 2 hours or so.  
Hope it helps. 

-----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: Monday, November 23, 2009 1:02 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 72, Issue 26

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

To subscribe or unsubscribe via the World Wide Web, visit
	http://lists.utsouthwestern.edu/mailman/listinfo/histonet
or, via email, send a message with subject or body 'help' to
	histonet-request <@t> lists.utsouthwestern.edu

You can reach the person managing the list at
	histonet-owner <@t> lists.utsouthwestern.edu

When replying, please edit your Subject line so it is more specific
than "Re: Contents of Histonet digest..."


Today's Topics:

   1. HT Opening (Marian Powers)
   2. cryojane tap transfer system (Richard Pattison)
   3. Re: cryojane tap transfer system (Merced M Leiker)
   4. Suggestions for Cassette Labeling (Fye Beth)
   5. RE: Suggestions for Cassette Labeling (Rathborne, Toni)
   6. Reference labs doing parafibromin? (Sebree Linda A)
   7. Mouse eyes (Durden, Kelley)
   8. Job Opening (Walzer Susan)
   9. Reponses to Merced and Richard Re: [Histonet] cryojane tape
      transfer	system  (gayle callis)
  10. RE: cryojane tap transfer system (CHRISTIE GOWAN)


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

Message: 1
Date: Sun, 22 Nov 2009 13:54:28 -0500
From: Marian Powers <mpowers <@t> dpspa.com>
Subject: [Histonet] HT Opening
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
	<5d7de0e60911221054i5971a474y64a461eeb5aa8cfc <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

DPS in Dover, Delaware, is currently seeking a full time histotech, day
or
evening shift.  All inquiries please email mpower <@t> dpspa.com



-- 
Marian L. Powers, HT(ASCP)
Manager, Technical Operations

Doctors Pathology Services
1253 College Park Drive
Dover, DE 19904
302-677-0000


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

Message: 2
Date: Mon, 23 Nov 2009 09:01:39 -0500
From: Richard Pattison <richard.pattison <@t> gmail.com>
Subject: [Histonet] cryojane tap transfer system
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <4B0A95C3.4090703 <@t> googlemail.com>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

Hi Everybody,
I was hoping to get some advice - I'm cryosectioning plant tissues and 
transferring sections to slides using the Cryojane system. However, i'm 
having problems in transferring the sections without them falling apart 
during the tape transfer. I'm fixing my tissue for 24 hours in 
ethanol:acetic acid (3:1), embedding in O.C.T, snap-freezing and then 
sectioning at between 2 and 14 microns. The sections seem to be ok but 
whenever i remove the adhesive tape from the slide a large part of the 
tissue is removed with it. As a result I lose the majority of my 
section. I've tried using both 1x and 1/2x slides (CFSA adhesive slides 
from instrumedics) but neither have given satisfactory results.
Does anyone have any suggestions as to how i could reduce the loss of 
tissue? Any advice would be much appreciated.
Thanks
Richard



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

Message: 3
Date: Mon, 23 Nov 2009 09:26:51 -0500
From: Merced M Leiker <leiker <@t> buffalo.edu>
Subject: Re: [Histonet] cryojane tap transfer system
To: Richard Pattison <richard.pattison <@t> gmail.com>,
	Histonet <@t> lists.utsouthwestern.edu
Message-ID: <5D6D90DC816B8E32D70F0BDD <@t> CDYwxp1931.ad.med.buffalo.edu>
Content-Type: text/plain; charset=us-ascii; format=flowed

Unless I'm missing something, I don't understand why people use this
tape? 
It seems like a marketing gimmick to me...ol' fashion' melting of
sections 
onto slides works perfectly for us...

?

Regards,
Merced

--On Monday, November 23, 2009 9:01 AM -0500 Richard Pattison 
<richard.pattison <@t> gmail.com> wrote:

> Hi Everybody,
> I was hoping to get some advice - I'm cryosectioning plant tissues and
> transferring sections to slides using the Cryojane system. However,
i'm
> having problems in transferring the sections without them falling
apart
> during the tape transfer. I'm fixing my tissue for 24 hours in
> ethanol:acetic acid (3:1), embedding in O.C.T, snap-freezing and then
> sectioning at between 2 and 14 microns. The sections seem to be ok but
> whenever i remove the adhesive tape from the slide a large part of the
> tissue is removed with it. As a result I lose the majority of my
section.
> I've tried using both 1x and 1/2x slides (CFSA adhesive slides from
> instrumedics) but neither have given satisfactory results.
> Does anyone have any suggestions as to how i could reduce the loss of
> tissue? Any advice would be much appreciated.
> Thanks
> Richard
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>



Merced M Leiker
Research Technician III
Cardiovascular Medicine
348 Biomedical Research Building
State University of New York at Buffalo
3435 Main St, Buffalo, NY 14214  USA
leiker <@t> buffalo.edu
716-829-6118 (Ph)
716-829-2665 (Fx)

No trees were harmed in the sending of this email.
However, many electrons were severely inconvenienced.




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

Message: 4
Date: Mon, 23 Nov 2009 08:38:05 -0600
From: Fye Beth <Beth.Fye <@t> HCAhealthcare.com>
Subject: [Histonet] Suggestions for Cassette Labeling
To: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	
<938F8EC5A524D34EB5796E23E52781D329A0FCA112 <@t> NADCWPMSGCMS05.hca.corpad.ne
t>
	
Content-Type: text/plain; charset="us-ascii"

There have been some great suggestions for reducing or catching slide
labeling errors.  I'm interested in the same for cassette labeling.
Currently, we have the patient's last name on one side of the cassette,
and the site listed on the other.  This makes it easier to identify
blocks if they are numbered incorrectly.

Beth A. Fye, CT (ASCP)
Pathology Technical  Manager
HCA Richmond Hospital Laboratories
office:  (804)228-6564
fax: (804)323-8638
<mailto:Beth.Fye <@t> hcahealthcare.com>






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

Message: 5
Date: Mon, 23 Nov 2009 09:41:14 -0500
From: "Rathborne, Toni" <trathborne <@t> somerset-healthcare.com>
Subject: RE: [Histonet] Suggestions for Cassette Labeling
To: "Fye Beth" <Beth.Fye <@t> HCAhealthcare.com>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	
<E78340C766A5284D999F5F5891DDF8900BAF8FCF <@t> smcmail.somerset-healthcare.co
m>
	
Content-Type: text/plain;  charset="utf-8"

That seems like a lot of effort. Do your pathologists do the grossing?
or PA's? 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Fye Beth
Sent: Monday, November 23, 2009 9:38 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Suggestions for Cassette Labeling


There have been some great suggestions for reducing or catching slide
labeling errors.  I'm interested in the same for cassette labeling.
Currently, we have the patient's last name on one side of the cassette,
and the site listed on the other.  This makes it easier to identify
blocks if they are numbered incorrectly.

Beth A. Fye, CT (ASCP)
Pathology Technical  Manager
HCA Richmond Hospital Laboratories
office:  (804)228-6564
fax: (804)323-8638
<mailto:Beth.Fye <@t> hcahealthcare.com>




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


CONFIDENTIALITY NOTICE
This message and any included attachments are from 
Somerset Medical Center and are intended only for the addressee.
The information contained in this message is confidential 
and may contain privileged, confidential, proprietary and/or 
trade secret information entitled to protection and/or exemption 
from disclosure under applicable law. Unauthorized forwarding, 
printing, copying, distribution, or use of such information is 
strictly prohibited and may be unlawful. If you are not the 
addressee, please promptly delete this message and notify 
the sender of the delivery error by e-mail or you may call 
Somerset Medical Center's computer Help Desk at 
908-685-2200, ext. 4050. 
-------------------------------------------------------------- 
Somerset Medical Center is the recipient of the 
2009 Orthopedic Surgery Excellence Award(tm) from HealthGrades,
the nation's leading health care ratings company.
 
Visit Somerset Medical Center's Web site 
- www.somersetmedicalcenter.com - 
for news, event listings, health information and more.

Join the Discussion: 
Facebook: www.somersetmedicalcenter.com/fb
Twitter: www.twitter.com/SomersetMedCtr

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

Message: 6
Date: Mon, 23 Nov 2009 09:35:07 -0600
From: "Sebree Linda A" <LSebree <@t> uwhealth.org>
Subject: [Histonet] Reference labs doing parafibromin?
To: "Histonet" <histonet <@t> pathology.swmed.edu>
Message-ID:
	
<8C023B4AB999614BA4791BAEB26E27381BF67E <@t> UWHC-MAIL01.uwhis.hosp.wisc.edu>
	
Content-Type: text/plain;	charset="us-ascii"

Good Monday morning,

One of our pathologists is getting pressure from the endocrinologists to
bring on Parafibromin (gene: hrpt2).  Are there any reference labs out
there offering this?

Thanks,

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




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

Message: 7
Date: Mon, 23 Nov 2009 10:44:21 -0500
From: "Durden, Kelley" <kelleydurden <@t> pathology.ufl.edu>
Subject: [Histonet] Mouse eyes
To: "'histonet <@t> lists.utsouthwestern.edu'"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<92E6B93E0A3D544C87DDDE33E7608AAE0956C7AD <@t> HSC-CMS01.ad.ufl.edu>
Content-Type: text/plain; charset="us-ascii"

My question concerns mouse eyes.

Can anyone send me their fixation suggestions?  We are receiving mouse
eyes that end up with a "wavy" look instead of a nicely defined "C" and
have no explanation for this occurrence.

Can anyone send me their mouse eye processing schedule?  We have a
protocol that has proved fast and true but would welcome other
suggestions.

Can you give me a good idea for making sure sections stick to slides
well?   Gold Plus?  What else?

Has anyone experienced a retinal detachment after or upon staining?
Retina attached after sectioning - detached after staining - just
routine H&E.  Should I heat for an extended period of time?

Kelley Durden, HT ASCP
University of Florida
Molecular Pathology Core


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

Message: 8
Date: Mon, 23 Nov 2009 09:48:19 -0600
From: Walzer Susan <Susan.Walzer <@t> HCAHealthcare.com>
Subject: [Histonet] Job Opening
To: "histonet <@t> pathology.swmed.edu" <histonet <@t> pathology.swmed.edu>
Message-ID:
	
<4BF03F5404EBDE409AF9232DA74B9DED2AEAB5E4B0 <@t> FWDCWPMSGCMS09.hca.corpad.ne
t>
	
Content-Type: text/plain; charset="us-ascii"

We have an opening for a tech, days no weekends at St Pete.General
Hospital in  St Pete,FL.  Call our HR dept at 727 384-1414 X 4814.


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

Message: 9
Date: Mon, 23 Nov 2009 10:20:58 -0700
From: "gayle callis" <gayle.callis <@t> bresnan.net>
Subject: Reponses to Merced and Richard Re: [Histonet] cryojane tape
	transfer	system 
To: "'Histonet'" <histonet <@t> lists.utsouthwestern.edu>
Cc: emmanuel.mineo <@t> leica-microsystems.com
Message-ID: <000101ca6c61$541e4d60$fc5ae820$@callis <@t> bresnan.net>
Content-Type: text/plain;	charset="us-ascii"

You wrote:

 

Unless I'm missing something, I don't understand why people use this
tape? 

It seems like a marketing gimmick to me...ol' fashion' melting of
sections 

onto slides works perfectly for us...

?

Regards,

Merced

 

Merced, 

Yes,  you are missing something.  If you have ever tried to cryosection
undecalcified bone or extremely difficult tissues that simply will not
result in "ol' fashion' melting" onto a slide , then you would
understand
why people use this unique cryosectioning system.  It is not some
"marketing  gimmick"  but an unique instrument  helping many
laboratories
obtain frozen sections that otherwise are scrunched up,  shattered, and
destroyed.  I suggest you go to the Instrumedics website or
www.alphelys.com
for a superb slide show  and learn how this instrument works before
making
assumptions about a technology that serves many of us more than well.  

 

A happy, informed user of the Cryojane................

 

Gayle M. Callis

HTL/HT/MT(ASCP)

Bozeman MT 

 

As for what Richard wrote: 

 

Hi Everybody,
I was hoping to get some advice - I'm cryosectioning plant tissues and 
transferring sections to slides using the Cryojane system. However, i'm 
having problems in transferring the sections without them falling apart 
during the tape transfer. I'm fixing my tissue for 24 hours in 
ethanol:acetic acid (3:1), embedding in O.C.T, snap-freezing and then 
sectioning at between 2 and 14 microns. The sections seem to be ok but 
whenever i remove the adhesive tape from the slide a large part of the 
tissue is removed with it. As a result I lose the majority of my 
section. I've tried using both 1x and 1/2x slides (CFSA adhesive slides 
from instrumedics) but neither have given satisfactory results.
Does anyone have any suggestions as to how i could reduce the loss of 
tissue? Any advice would be much appreciated.
Thanks
Richard
 
Dear Richard, 
 
I could be the fixative you are using that causes the problem.   If the
fixative contains alcohol, the alcohol acts as antifreeze when you try
to
snap freeze a tissue, animal or plant.  The alcohol may cause problems
with
how the pink tape sticks to the face of the plant tissue, and allows
them to
fall apart during the tape transfer.   If you rinse away the fixative,
then
you should cryoprotect the fixed plant tissue with 30% sucrose before
snap
freezing. vbThis will remove the alcohol.  If cryoprotection causes
problems
with the final staining results, then try unfixed plant tissue, snap
freeze,
Cryojane tape transfer the section and then fix the transferred plant
section in  your favorite fixative.   You may have to optimize the time
in
fixative though.   
 
Other suggestions:  
 
Do a double UV flash, but wait for 15 to 20 seconds between flashes.
You
must allow the UV light source (capacitor) build up enough charge to
work
properly.  This double flash seems to help polymerize the coating more
thoroughly, and the section should transfer better.  Also, the tape must
be
removed at an angle across the slide,  very slowly, and inside the
cryostat
(I am sure you probably do this already.) 
 
Also, try the 4X slide if you still have problems  with 1/2X and 1X
slides.
You might ask Leica to send you a few to try before investing in a whole
box
of these.  Contact Emmanuel Mineo, Intrumedics Product Manager
emmanuel.mineo <@t> leica-microsystems.com for the 4X slides.  Manny is a
nice
gentleman who has worked with Cryojane for many years and has always
been
helpful to us.   Once again, do the double UV light flash with the 4X
slides. They are gooey, but may/should hold more securely.  
 
With undecalcified bone, we use the 1/2X but do the double flash
routinely
for all sections.   
 
Good luck
 
Gayle Callis    
 
 


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

Message: 10
Date: Mon, 23 Nov 2009 17:22:40 +0000
From: CHRISTIE GOWAN <christiegowan <@t> msn.com>
Subject: RE: [Histonet] cryojane tap transfer system
To: <leiker <@t> buffalo.edu>, <richard.pattison <@t> gmail.com>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <SNT122-W392B7D2052613E9C485796AE9E0 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"



 Hi Richard,

I am assuming that you are using the flash in the cryojane system to
adhere the specimen to the slide. Make sure your tape is not too old and
don't store it in the cryostat. It sounds like you are able to get the
section but unable to get it to adhere. Make sure you pass your roller
over the tape and slide several times before putting it under the flash
and always use a positive charged slide from a freshly opened box. Good
luck. I hope this helps a little

Christie
 		 	   		  

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

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

End of Histonet Digest, Vol 72, Issue 26
****************************************
Notice:  This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system.