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 37°C
4. best protocol for RNA extraction?
RNeasy Mini kit from QIAGEN?
All suggestions and in-house knowledges are welcome.....
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