Working Procedures in Microwave Histology

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by Vincent R. Klump, Jr., HT (ASCP), Histology Services, East Haven, CT

note: Mr. Klump was instrumental in the development of a standardized, consistent, reproducible protocol for microwave histoprocessing in the original Energy Beam Sciences H2500 Microwave Processor.
Contents:

Microwave Tissue Processing
Antigen Retrieval
Special Stains
Microwave Stimulated Decalcification
Microwave Stimulated Fixation
Slide Drying in the Microwave
Microwave Tissue Processing QC Chart
Microwave Tissue Processing

Tissue criteria: Maximum tissue size for this procedure: 3mm x 3mm x 3mm. Variations of this size can also be accommodated, but total tissue volume should not exceed 3mm cubed.

Examples: 3mm punch biopsies, core needle biopsies, core bone marrow biopsies, shave biopsies.

Tissue should be fixed in 10% NB Formalin for a minimum of four hours.
Equipment:

* Laboratory microwave equipped with a temperature probe accurate to +/- 1篊
* Microwave-safe container holding a minimum of 250ml
* (optional) microwave-safe cassette rack supplied by Energy Beam Sciences
* 100% ethanol
* 100% isopropanol
* liquid paraffin
* boiling chips (preferably, common stones not bigger than 1cm x .5cm x .5cm)

Procedure:

1. Rinse tissue for 5 minutes in running water
2. dehydrate in 100% ethanol for 15 minutes at 65篊 in microwave
3. clear in 100% isopropanol for 10 minutes at 74篊 in microwave
4. infiltrate (3 steps) in liquid paraffin for 5 minutes at 65篊 in microwave with boiling chips, 5 minutes at 74