Enhancement of in Situ Hybridization Signal with Microwave Heating

Enhancement of in Situ Hybridization Signal with Microwave Heating 

Fix human tissues with 4% paraformaldehyde in PBS for 24 hr, embed in paraffin, and cut 4- to 7-p.m-thick sections. 

Mount sections on silane-treated glass slides. 

Deparaffmize with three changes of 5min each in xylene, followed by three changes of 100% ethanol, two changes of 95% ethanol, and two changes of 75% ethanol. 

Place slides in microwave transparent jars containing 0.01 M sodium citrate buffer (pH 6.0), and heat to the boiling point at maximum power (700 W) in a microwave oven equipped with a rotating plate cover the jars with loosely fitting lids unless 2-3 cm empty space is present above the buffer level in the jar. 

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