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