The use of UV light as a cross-linking agent for cells and tissue sections in in situ hybridization.
UV cross-linking is introduced as a novel method to stabilize tissue on microscopic slides and to immobilize target molecules in biological material for in situ hybridization. UV illumination dramatically improves the stability of tissue sections and isolated cells on slides coated with gelatin/poly-lysine. Even during prolonged high-stringency washes, specimens remain firmly attached to the support layer. At the same time, the signal intensity is increased significantly whereas background levels remain as low as without UV illumination. These results indicate that while target RNAs and other molecules in the biological material are covalently cross-linked with their nearest neighbor molecules, tissues nonetheless remain penetrable, and target molecules remain accessible. We expect that this simple and efficient technique will find widespread applications in in situ hybridization methodology.
Tiedge
H
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Fishberg Research Center for Neurobiology, Mount Sinai School of Medicine, New York, NY 10029.
D000818:Animals; D003432:Cross-Linking Reagents; D019286:Endopeptidase K; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000528:radiation effects; D015347:RNA Probes; D051381:Rats; D012697:Serine Endopeptidases / Q000494:pharmacology; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D015347:RNA Probes; D012313:RNA; D012697:Serine Endopeptidases; D019286:Endopeptidase K