Standard processing for IEM

1. Fixation:
3% PFA in 0.1M Buffer with 0.1% GA and 4% sucrose

2. Wash:
4x15 min. with correlative buffer containing 0.1M Sucrose @ 4C
1x30 min. with correlative buffer containing 0.1M Sucrose and 0.1M Glycine @4C

3. Dehydration with METOH:
30% 1x15 min. @4C
50% 1x15 min. @4C
70% 1x15 min. @-10C
85% 1x15 min. @-20C
95% 1x15 min. @-20C
100% 3x15 min @-30C

6. Infiltration with Lowicryl LK4M:
METOH:LK4M = 1:1 for 1 hr. @-30C
METOH:LK4M = 1:2 for 1 hr. @-30C
METOH:LK4M = 1:3 for 1 hr. @-30C
Pure LK4M 1 hr. @-30C
Pure LK4M overnight @-20C

7. Embedment:
Embed into Beem Capsules and cure under UV in a CO2 environment @-35C for 36 hr. and under UV in a CO2 environment for 12 hr. @ RT.