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PELCO BioWave® Pro+ Microwave Tissue Processor
Electron Microscopy Tissue Processing Application Kit

Microwave Protocol for Electron Microscopy

Microwave Procedure by CHARLES K. MESHUL, PH.D.
Professor, Department of Behavioral Neuroscience and Pathology (OSHU)
Research Biologist and Director, Electron Microscopy Facility (VA)

Step Description User
Prompt		 Time
(hr:min:sec)		 Watts Temp Repetitions Vacuum
1 Osmium On 00:13:00¹ 100 60 1 Cycle
2 Rinse On 0:00:40 150 60 2 Off
3 UA On 00:06:00² 100 60 1 Cycle
4 50% ETOH On 0:00:40 150 60 1 Off
5 75% ETOH On 0:00:40 150 60 1 Off
6 95% ETOH On 0:00:40 150 60 1 Off
7 100% ETOH On 0:00:40 150 60 2 Off
8 Propylene Oxide On 0:00:40 150 60 1 Off
9 1:1 PO/Resin On 0:03:00 150 60 1 Cycle
10 100% Resin On 0:03:00 150 60 3 Cycle
IF TISSUE IS HAND CUT OR THICKER THAN 100µm CONTINUE WITH THE FOLLOWING STEPS
11 100% Resin On 0:03:00 150 60 2 Cycle
place tissue in molds or flat embed using ACLAR films place in 60°C overnight.
if using for post embedding immunoreactivity do not leave in the microwave longer than 20hrs
¹ 3 MIN WATTS ON / 2 MIN WATTS OFF / 3 MIN WATTS ON / 2 MIN WATTS OFF / 3 MIN WATTS ON
² 2 MIN WATTS ON / 2 MIN WATTS OFF / 2 MIN WATTS ON

Solutions:
Osmium: 1:1 of 2% Osmium Tetroxide in H20 to 3% Potassium Ferricyanide in H20
Uranyl Acetate (UA): 0.5% Uranyl Acetate in H20