Table 1.
Optimization of sample preparation for TEM and cryo-ultramicrotomy.
| Method | Buffer | Fixation | Post-fixation | Resin |
|---|---|---|---|---|
| Procedure 1 (TEM) | Cacodylate + 4% sucrose | 3.0% formaldehyde, 0.035% glutaraldehyde, 4% sucrose and 0.075% ruthenium red, 0.075% L-lysine acetate | 1% osmium tetroxide; 0.075% ruthenium red | LR-White |
| Procedure 2 (TEM) | PBS + 4% sucrose | 4.0% formaldehyde, 1.25% glutaraldehyde, 0.01 M CaCl2, 4% sucrose and 0.075% ruthenium red | 1% osmium tetroxide; 0.075% ruthenium red; | Epon-Araldite |
| Procedure 3 (TEM) | Cacodylate | 4.0% formaldehyde, 1.25% glutaraldehyde, 0.01 M CaCl2 and 0.075% ruthenium red, 0.075% L-lysine acetate | 1% osmium tetroxide; 0.075% ruthenium red | Epon-Araldite |
| Procedure 4 (TEM) | Cacodylate + 4% sucrose | 4.0% formaldehyde, 1.25% glutaraldehyde, 0.01 M CaCl2, 4% sucrose and 0.075% ruthenium red, 0.075% L-lysine acetate | 1% osmium tetroxide; 0.075% ruthenium red | LR-White |
| Procedure 5 (TEM) | Cacodylate + 4% sucrose | 4.0% formaldehyde, 1.25% glutaraldehyde, 0.01 M CaCl2, 4% sucrose and 0.075% ruthenium red, 0.075% L-lysine acetate | 1% osmium tetroxide; 0.075% ruthenium red | Epon-Araldite |
| Procedure 6 (Cryo-ultramicrotomy) | Cacodylate + 4% sucrose | 4.0% formaldehyde, 1.25% glutaraldehyde, 0.01 M CaCl2, 4% sucrose and 0.075% ruthenium red, 0.075% L-lysine acetate | N/A | N/A |