TEM MICROWAVE TISSUE PROCESSING PROTOCOL
A) Check water levels in load coolers.
B) Turn on the load cooler pump prior to microwave use.
C) Heat the water in the microwave (2 min on HIGH).
Step Temp. Processing Time Notes
1) Fixation 37 1 or 2 X 40 sec. 600 ul in eppendorf
Cool to 20 on ice
Probe in load cooler
2) Buffer rinse RT 2 X 2.5 min.
3) Osmium fix 37 1 X 40 sec. Cool to 20 on ice
Probe in load cooler
4) Buffer rinse RT 1 X 30 sec.
5) Dehydration (Acetone or Ethanol) Transfer to baskets
50% 45 2 X 40 sec. Baskets in petri dish
Fill petri 3/4 full
70% 45 2 X 40 sec. Probe in sol'n 4 mm
Change the water in
90% 45 2 X 40 sec. the 2nd beaker
when it's hot to the
100% 45 3 X 40 sec. touch.
6) Infiltration
1 : 1 50 1 X 15 min. Probe in dish
Change the water in
100% resin 50 3 X 10 min. the 2nd beaker
when it's hot to the
touch.
7) Polymerization
95 1 X 15 min. Use BEEM capsules
none 1 X 60 min. Put parafilm under
caps to seal shut