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TEM - Embedding

Standard embedding

The aim of the embedding procedure is to produce blocks that are suitable for ultra thin sectioning and still preserve the fine structure with as little alteration as possible. The procedure starts with fixation of the specimen, usually  a double fixation with an aldehyde and osmium tetroxide in buffer. After washing, with buffer, the specimens are dehydrated in a graded series of ethanol or acetone. The solvent is then replaced by a monomeric resin (usually an epoxy resin) which is polymerized into a hard block that can be sectioned with an ultra microtome into 50- 80 nm slices.

This procedure is suitable for tissue and large particles (usually bacteria and organelles).

Embedding for immunolabeling

Antigens in an embedded specimen can often be localized with a complex of colloidal gold and antibody if the antigenesity is preserved, since the structures are protruding out of the section surface. To preserve the antigenesity of the specimen the above procedure is altered somewhat. The fixation is milder, only aldehyde, and the epoxy resin is replaced with a methacrylate. The whole procedure is usually performed at low temperature, -50 – 0 degrees.

Kidney

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Cilier

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Small bacteria

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