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 H
20 to 3% Potassium Ferricyanide in H
20
Uranyl Acetate (UA): 0.5% Uranyl Acetate in H
20