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.
Quentin
Thomas
T
Department of Pediatric Cardiology and Pediatric Intensive Care Medicine, Georg-August University, D-37075 Goettingen, Germany.
Poppe
Andrea
A
Bär
Karin
K
Sigler
Albrecht
A
Foth
Rudi
R
Michel-Behnke
Ina
I
Paul
Thomas
T
Sigler
Matthias
M
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