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

[Karl Brand]

Histonetters,

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!

yours,

Karl

Karl Brand k.brand <@t> 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