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
<pru...@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-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of gayle
callis Sent: Monday, November 23, 2009 10:21 AM
To: 'Histonet'
Cc: emmanuel.mi...@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.mi...@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
lei...@buffalo.edu
716-829-6118 (Ph)
716-829-2665 (Fx)
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However, many electrons were severely inconvenienced.
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