in situ hybridization

Vasundhara Kandachar kandachar at
Thu Jun 28 13:34:59 EST 2001


We are trying to prepare sections of spleen, thymus
and lymphnodes from mouse for in situ hybridization. 
We find holes in the tissue sections.  We are thinking
that it could be infiltration problem.  

We processed the tissue as follows:
Fixation: 4% paraformaldehyde in PBS overnight (RT)
Dehydration:  5 minutes in each of six different
concentrations of ethanol at RT
Used xylene as a solvent for paraffin infiltration.  
Paraffin Infiltration: for 10 days at 60 degrees C.

If anybody could suggest modification in the above
method of tissue processing, please let us know.

Thanks in advance.

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