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Microwave Extended Mini Protocol

The following is a modified protocol for fixing, dehydrating, and infiltrating large animal samples or "difficult" plant samples (hard or dense materials).

• Samples can be 3–5mm on a side.
• The MRA BP111RS oven requires no water load.
• Power setting = 55.
• The temperature of the water bath (pyrex dish into which the sample vials sit during MW) should be equal to the infiltration solution.
• Water bath volume appx 0.9l, with 12 sample vials in a 9x9" pyrex dish.

step	time (min)	temp	Bath temp
10% Formalin (4%PFA in buffer)	30	37	4
50% EtOH	30	67	RT
95% EtOH	30	67	RT
100% EtOH + eosin	10	67	RT
50% EtOH:50% Isop	30	77	RT
100% Isopropanol	30	77	RT
50% Isop:50% wax	30	77	60
100% paraffin wax	90	67	60
100% paraffin wax	90	67	60

Samples should be embedded (cast) immediately at the end of the last wax step.

For Animal tissue, staining with H&E yielded good results.

S Ruzin, J. Nemson

8/10/05