Table 1 Tissue preparation and processing methods for electron microscopic imaging

1

Postfixed with 4% paraformaldehyde +2.5% glutaraldehyde in 0.1M PB for 2 weeks at 4°C

Postfixed with 4% paraformaldehyde +2.5% glutaraldehyde in 0.1M PB for 14 hours at 4°C

2

Vibratome section at 80 μm, store in 0.1M PB

Washed and stored in 0.1M PB

3

Osmification in OsO₄ 0.1M PB for 20 min and washed in 0.1M PB

4

Dehydration in graded ethanol solutions and 100% acetone

5

1:1 acetone/Araldite for 1 hour at room temperature and in pure Araldite overnight at 4°C

6

Flat embedding on siliconized slides with Araldite

7

Polymeriztion at 60°C for 48 hours in oven

8

Dissection of white matter from embedded sections and glue to blank resin blocks

9

Thin sectioning at 70-80 nm and stain grids with uranyl acetate and lead citrate solutions

10

Examination of ultrathin sections on electron microscope, image processing and quantification