[Histonet] quickest freezing method AND maintain morphology of mouse brains for LCM then microarray???

From:"Karl Brand"


Like to hear your methods for the most rapid method of freezing tissue for
subsequent cryosectioning, whilst maintaining best possible morphology.

We using the PALM LCM scope to capture a small region of mouse brain, from
fresh, rapid heamatoxylin stained ~8um cryosections. After this microarray
analysis will be performed, so-

**minimising RNA degradation whilst maintaining morphology are the critical
goals here**

Until now i have dropped the brain into TissueTek OCT medium and freezing on
a bed of dry ice which typically takes ~4 mins or so. The morphology is
satisfactory using this method, but working with such small amounts of
tissue makes every second critical i feel. I just tried dropping the
specimen (in OCT in mould) into isopentane, but a big crack formed in the
OCT which i fear may also cause a fissure in the specimen!

Suggestions? Experience? Greatly appreciated!



Karl Brand k.brand@erasmusmc.nl
Department of Genetics
Erasmus MC
Dr Molewaterplein 50
3015 GE Rotterdam
lab +31 (0)10 408 7409 fax +31 (0)10 408 9468 mob +31 (0)621 446 504

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