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
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