1 / 4

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.

lisandra-graham
Download Presentation

Ultramicrotomy

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  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

More Related