RE: Problems cryosectioning skin
Hi Nicola,
I've been cutting skin for a very long time, so here are some of my thoughts.
The advice from Greg is great and I do all those things. I find it easier to
cut so that I'm cutting both epidermis and dermis at the same time, but Louise
doesn't, so my advice is if it doesn't cut one way try another. Whatever works
for you is the best way! I cut at -24-25, and I think maybe the cracking of
your block may be because the temperature is too low. You may also get
separation if your sample is very wet when you embed if you are embedding fresh
(not frozen tissue). Unfortunately I've also had it happen putting frozen
tissue in room temperature OCT, so I cool the OCT.
And although this won't help you now, try to purchase a CryoJane. It has made
my life much much easier. Although it may appear costly to buy and buy the
slides and tape, consider that you will have less section loss from curling,
etc and having sections come off during staining, therefore less time spent
cutting. (I'm not related, don't get a commission or perks from Instrumedics,
by the way)
-----Original Message-----
From: louise renton [SMTP:louise_renton@hotmail.com]
Sent: Tuesday, May 27, 2003 3:09 AM
To: n.cragg@epistem.co.uk; histonet@pathology.swmed.edu
Subject: Re: Problems cryosectioning skin
Dea Nicola
Could this be an orietation problem? I always found that it was easier to
section skin when the tissue was (a) orientated so that the epidermis was
cut last as the blade/knife passed through the tissue and (b) if the tissue
was angled slightly so that the minimum area was presented to the cutting
edge.
Finally, although this might bring down a cascade of criticism : try warming
the block slightly with a suitably gloved finger before taking each section
best regards
Louise Renton
Bone Research Unit
MRC
Johannesburg
South Africa
Tel & fax +27 11 717 2298
"Time flies like an arrow, fruit flies like a banana"
----Original Message Follows----
From: ncragg
To: 'HistoNet Server'
Subject: Problems cryosectioning skin
Date: Mon, 26 May 2003 11:29:35 +0100
Please can someone advise me on how to cut good cryosections of skin? I am
having real trouble getting good mouse sections, the tissue tears and
separates from the OCT as I'm cutting it. I've tried changing the
temperature, from -20 to -15, as the manual for the cyrostat seem to be
saying the temperature was too cold, changing the clearance angle, new
blades, changing the speed - I'm exasperated. I know it does take a lot of
practice to get the technique right, I learnt this with paraffins, but I
really don't know what I'm doing wrong. Please help!! I have 200 samples to
cut!!!
Yours desperately,
Nicola Cragg
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