A novel method for processing resin-embedded specimens with metal implants for immunohistochemical labelling.
A major technical problem in the processing of resin-embedded tissues is the adhesion of the tissue sample on glass slides for immunohistochemical labelling. We therefore established a novel protocol for processing such specimens with improved attachment of the tissue sample during resin removal (deplastification). In order to demonstrate the feasibility of the procedure we employed a panel of smooth muscle cell maturation markers. The technique makes use of a silicone glue (Elastosil E41; Wacker Chemie, München, Germany) to attach the tissue samples to the glass slides. This allows resin dissolution in xylene/2-methoxyethylacetate without detachment of the sample from the slide. Our results demonstrate successful immunohistochemical labelling with primary antibodies directed against: smooth muscle actin, smooth muscle myosin, h-caldesmon, desmin, vimentin and von Willebrand factor. In conclusion, we have established a new and successful method for resin-embedded sample adhesion on glass slides. The developed protocol is feasible for investigation of cells which are involved in intimal proliferation following stent implantation.
Department of Pediatric Cardiology and Pediatric Intensive Care Medicine, Georg-August University, D-37075 Goettingen, Germany.
Thomas
T
Quentin
Andrea
A
Poppe
Karin
K
Bär
Albrecht
A
Sigler
Rudi
R
Foth
Ina
I
Michel-Behnke
Thomas
T
Paul
Matthias
M
Sigler
D005898:Glass / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D010232:Paraffin / Q000737:chemistry*; D012826:Silicone Elastomers / Q000737:chemistry*; D015607:Stents; D013499:Surface Properties; D014470:Umbilical Cord / Q000166:cytology
D012826:Silicone Elastomers; D010232:Paraffin