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Immuno-labelling for TEM cryo-chambertemp. control liquid nitrogen sectioning control Sample processing 1.Fix 4% PFA 2.Wash/Quench 3.Harvest cells/tissue.

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Presentation on theme: "Immuno-labelling for TEM cryo-chambertemp. control liquid nitrogen sectioning control Sample processing 1.Fix 4% PFA 2.Wash/Quench 3.Harvest cells/tissue."— Presentation transcript:

1 Immuno-labelling for TEM cryo-chambertemp. control liquid nitrogen sectioning control Sample processing 1.Fix 4% PFA 2.Wash/Quench 3.Harvest cells/tissue 3a.Gelatin embedding 4.2.3M sucrose infusion 5.Cut ≤1mm 3 pieces 6.Mount on cryo-stub 7.Snap freeze liquid N 2 http://www.liv.ac.uk/emunit

2 1.2.3. 4. 5.Immuno-labelling 6.MC/UA contrasting 7.Grid looping 8.Drain excess 9.Dry Immuno-labelling for TEM 1.Trim 2.Section 3.Pick-up 4.Add to EM grids http://www.liv.ac.uk/emunit

3 Immuno-labelling for TEM 5.6. Quench BlockWash 1°Ab 50µl washes 5µl/grid Ab incubations 5.Immuno-labelling 6.MC/UA contrasting 7.Grid looping 8.Drain excess 9.Dry 1.Trim 2.Section 3.Pick-up 4.Add to EM grids http://www.liv.ac.uk/emunit

4 7.8.9. Immuno-labelling for TEM 5.Immuno-labelling 6.MC/UA contrasting 7.Grid looping 8.Drain excess 9.Dry 1.Trim 2.Section 3.Pick-up 4.Add to EM grids http://www.liv.ac.uk/emunit

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