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Ultramicrotomy

3mm diamond. Ultramicrotomy. Sections: 50-70 nm thick 0.1-0.5 mm square. Knives Glass Diamond. Resin good morphology. Cryo good immuno-labelling. http://www.liv.ac.uk/emunit. Ultramicrotomy. Sections: 50-70 nm thick 0.1-0.5 mm square. Knives Glass Diamond. specimen. knife.

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Ultramicrotomy

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  1. 3mm diamond Ultramicrotomy Sections: 50-70 nm thick 0.1-0.5 mm square Knives Glass Diamond Resin good morphology Cryo good immuno-labelling http://www.liv.ac.uk/emunit

  2. Ultramicrotomy Sections: 50-70 nm thick 0.1-0.5 mm square Knives Glass Diamond specimen knife Resin good morphology Cryo good immuno-labelling speed/thickness controller http://www.liv.ac.uk/emunit

  3. Processing for resin sectioning Sample: eg. cells in 1 x 6cm dish. Final pellet ≤1mm3 Protocol: Fix (4% paraformaldehyde, 2.5% glutaradehyde) 4hrs-o/n Wash and quench (50mM glycine) 5 min 1% OsO4 1.5hrs 5% alcoholic uranyl acetate 1.5hrs Dehydrate (50%, 70%, 90%, 100% ethanol, 100% acetone) 1.5hrs Infiltrate (araldite resin; 30:70, 70:30, 100%) 4 hrs Polymerise 60ºC o/n http://www.liv.ac.uk/emunit

  4. Processing for resin sectioning 1. Trim block (≤0.3mm2 face) 2. Sectioning 50-70nm (silver/gold colour) 3. Collect sections on EM grid http://www.liv.ac.uk/emunit

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