[Histonet] help

From:Conny Gysemans

Dear histonetters,

We are a small lab in Belgium trying to start with laser capture 
microdissection on pancreatic tissue.  We would use the Leica LMD system 
which uses mounting of sections on frame foil membranes (membranes sealed 
around the edges with nail varnish).

We would like to use Mayers' hematoxylin - eosin staining to identify both=20
infiltrating immune cells and endocrine cells in the pancreatic islets.

Can anyone give us some ideas on how to proceed?
1. preparation of tissue (fixation?)
2. applying specimen on slides (can tissue adhesive be used if we want to 
do RNA extraction thereafter?)
3. which protocol for staining?
	30 sec 100% ethonol (3x)
	30 sec 96% ethanol (1x)
	30 sec 70% ethanol (1x)
	30 sec distilled water (1x)
	1 min hematoxylin
	rinse in water
	1 min eosine
	rinse in water (remove excess water by placing a filter on the sample)
	30 min air dry at 37C
4. best protocol for RNA extraction?
	RNeasy Mini kit from QIAGEN?

All suggestions and in-house knowledges are welcome.....

Best regards 


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