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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
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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
![Page 3: Immuno-labelling for TEM cryo-chambertemp. control liquid nitrogen sectioning control Sample processing 1.Fix 4% PFA 2.Wash/Quench 3.Harvest cells/tissue](https://reader036.vdocuments.site/reader036/viewer/2022070409/56649e915503460f94b96f38/html5/thumbnails/3.jpg)
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
![Page 4: Immuno-labelling for TEM cryo-chambertemp. control liquid nitrogen sectioning control Sample processing 1.Fix 4% PFA 2.Wash/Quench 3.Harvest cells/tissue](https://reader036.vdocuments.site/reader036/viewer/2022070409/56649e915503460f94b96f38/html5/thumbnails/4.jpg)
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