Immuno-labelling for TEM

cryo-chamber temp. control

liquid nitrogen

sectioning

control

Sample processing1. Fix 4% PFA2. Wash/Quench3. Harvest cells/tissue3a. Gelatin embedding4. 2.3M sucrose infusion5. Cut ≤1mm3 pieces6. Mount on cryo-stub

7. Snap freeze liquid N2

http://www.liv.ac.uk/emunit

1. 2. 3.

4.

5. Immuno-labelling6. MC/UA contrasting

7. Grid looping8. Drain excess9. Dry

Immuno-labelling for TEM

1.Trim2.Section3.Pick-up4.Add to EM grids

http://www.liv.ac.uk/emunit

Immuno-labelling for TEM

5. 6.Quench

Block Wash1°Ab

50µl washes5µl/grid Ab incubations

5. Immuno-labelling6. MC/UA contrasting

7. Grid looping8. Drain excess9. Dry

1.Trim2.Section3.Pick-up4.Add to EM grids

http://www.liv.ac.uk/emunit

7. 8. 9.

Immuno-labelling for TEM

5. Immuno-labelling6. MC/UA contrasting

7. Grid looping8. Drain excess9. Dry

1.Trim2.Section3.Pick-up4.Add to EM grids

http://www.liv.ac.uk/emunit