[Histonet] Another quick sectioning question

[Lewis, Patrick]

When I was cutting my block I noticed some ribboning of the tissue.  Is
there a known cause (or several) for this.  I am wondering if my
microtome blade was too tight or too loose or if my block froze poorly,
or if I just needed to reorient my block .  I was cutting 7 um at -20.
Its human tonsil,  It's been a long time since I used a cryostat (we're
using a leicaCM3000).    The ribboning almost disappeared when I changed
blades after cutting about 30 sections into the block.  But I still had
some ribboning of the tissue even though I was getting pretty clean OCT
slices away from the tissue.

 

Also, anyone have advice on disinfecting cryostats.  I will eventually
be cutting infectious tissues, HIV human samples and NHP macaques.  We
plan to follow Leicas disinfection procedure in which you bring the
cryostat to Room temp then spray with 5% Bleach let it sit for 10-20
minutes then wipe and spray with 70% ethanol and sit for 10 min then
wipe and spray with 100% ethanol. And drain and wipe.  Anyone have
advice on cleaning and if it's necessary to remove any portions of the
cryostat to facilitate cleaning.  I figure it will be a two to three day
process. To bring it to room temp, clean it, then bring it back down to
-20.  My main concerns are making sure its disinfected, and then making
sure we remove all the bleach so that we don't get corrosion.  

 

Thanks for all your help and advice.

 

Patrick.

 

Patrick Lewis

Research Associate II-Bench| Infections and Prematurity

Seattle Children's Research Institute

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patrick.lewis <@t> seattlechildrens.org

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