SPAM-LOW: Reponses to Merced and Richard Re: [Histonet] cryojane tape transfer system

Wed Dec 9 10:28:48 CST 2009

I guess I did not realize what ongoing discussion my comment about the 
CryoJane tape transfer would spark! Thanks to the replies I have received 
in private and in public, I see that it indeed helps one make lovely bone 
sections.

In the past 9 years that I have cryosectioned, I have done a range of soft 
tissues, but never bone. CryoJane had been pushed on me a time or two by a 
rep to use on my (non-bone) tissues. I discovered, however, that it was not 
necessary on my (non-bone) tissues. I was never informed of using it on 
bone (until now!), never even crossed my mind, since I'd never done bone.

So, I'm sorry for anyone who may have been offended by my question about 
the use of the CryoJane tape transfer.  I indeed figured I had to be 
missing something!   :-)

Regards,
Merced

--On Sunday, December 06, 2009 2:31 PM -0700 Patsy Ruegg 
<pruegg <@t> ihctech.net> wrote:

> Here, here Gayle, I have a picture on my website of a calcified horse
> carpal bone I cut using the Instrumedics tape transfer system, I bet no
> one would have been able to do that without using the tape.
> www.ihctech.net
>
>
>
> Patsy Ruegg, HT(ASCP)QIHC
> IHCtech
> 12635 Montview Blvd. Ste.215
> Aurora, CO 80045
> 720-859-4060
> fax 720-859-4110
> www.ihctech.net
> www.ihcrg.org
>
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of gayle
> callis Sent: Monday, November 23, 2009 10:21 AM
> To: 'Histonet'
> Cc: emmanuel.mineo <@t> leica-microsystems.com
> Subject: SPAM-LOW: Reponses to Merced and Richard Re: [Histonet] cryojane
> tape transfer system
>
> 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
>
>
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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)

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