[Histonet] embedding multiple snap frozen brains for cryosectioning

From:Zsuzsa Penke-Verdier

Hello Netters,

For an in situ hybridation experiment, I need to cut snap frozen mouse 
brain tissue with a cryostat and put several sections (minimum 7, each 
from a different brain) on the same slide. Because mRNA is rapidly 
degraded if cutting takes too long, I cannot afford to cut each of the 
7 brains individually. This is why I need to make a composite block out 
of several brains. However, making such a block is problematic for 2 
reasons :

1.	It is important that each brain section should be in the same 
relative plane (the same anteroposterior coordinate). Does anyone have 
an idea how I could achieve this ?

2.	How could I avoid thawing of brains occasioned by adding freezing 
medium ?

Any suggestion would be welcome. Thanks in advance for your help !

Zsuzsa Penke-Verdier
Post doc researcher

Laboratory of the Neurobiology of Learning, Memory and Communication
NAMC, CNRS UMR 8620, Bāt 446
University Paris-Sud
91405 Orsay Cedex, FRANCE
Tel:  33 (0)1 69 15 49 96
Fax: 33 (0)1 69 15 77 26

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