|From:||Gayle Callis <email@example.com>|
Ready for the lecture?!?
How do you apply tape? I do frozens exclusively, no problems, an
occasional glitch, but attribute that to technician error, impatience.
I like a block shaped so the tape fits over a ledge of OCT, with narrowest
edge of tissue facing down, or will be cut first, the path of least
resistance to knife edge, an inverted triangle of sorts.
My chamber temp for soft tissues is set at -26C in order for UV unit to
maintain cold temps. The block face needs to have OCT surrounding tissue,
and tape fits on face with bit of overlap. Peel tape apart carefully, I
try to not touch tape or bend it in any way. Approach block face so tape
adheres smoothly to surface, then roll so there are NO BUBBLES under tape,
takes firm pressure, and I try to NOT roll OVER edges. One can learn to
apply tape so bubbles are not there, but you can see them.
Roll back and forth several times, pick up tape at bottom to hold above
blade, BUT DO NOT PULL UP ON TAPE NOR LIFT UP ON IT WHILE CUTTING. It will
feed toward you at approximately parallel to same angled shape of knife
holder. Tape should over knife edge without detachment from block NOR
sticking to knife holder. You need to let the microtome do the work, once
you grasp tape and turn flywheel, the section/tape should feed towards you,
just as in any frozen section, smoothly, without hesitation in cutting.
Don't go too fast, don't go to slow, do the baby bear thing, just right!
Make sure roller is CLEAN, garbage adhering prevents 100% contact with tape
to block face.
I lay mine inside chamber so it does not touch anything but tape when used,
clean it with alcohol after use, and store it in refrig until needed, often
in a baggie.
Also, make sure you are sitting in a very comfortable position and not
straining your arms, back, anything, this makes tape application a tense,
difficult manuever (sp?). I have a pneumatic chair that raises and lowers
for the short, medium, tall people - comfort is mandatory. Adjust height
so you feel relaxed when you do tape thing, it is much easier.
Enough of this, I love Cryojane for my bone, lung, any difficult tissues,
with really fatty breast, you need to lower the temp even more, but -26C
might work. I am not sure what you cut, we do not use it for everything.
Some huge chunks of bovine lung were cut, not a problem, and I mean HUGE!
Impressed the heck out of faculty member who always cut lung looking like
overhanging sand dunes, and you cannot perfuse a bovine lung with OCT
easily. Would take gallons of the stuff!
e. At 01:52 PM 5/22/01 -0500, you wrote:
>hello all, to add to Linda Jenkins question. I am having the same problem
>with the tape not adhering to the block face of my frozen section. We very
>recently purchased an Instrumedics CryoJane tape transfer system. Any
>pointers/tips will be much appreciated in remeding this problem ASAP.
>Atoska S. Gentry B.S., HT(ASCP)
>Research Assistant II
>Scott-Ritchey RSCH Center
>Auburn University, AL 36849
>PH: (334) 844-5579
>Fax: (334) 844-5850
Veterinary Molecular Biology
Montana State University - Bozeman
Bozeman MT 59717-3610
404 994-4303 (FAX)
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