Cytology, General Histology and Embryology - V. K. Naphanyuk 2002

Methods of Histological Research
Post-Vital Research Methods
Features of Sample Preparation for Electron Microscopic Studies

1. Sampling and fixation of Tissue and organ specimens (approximately 1 mm3 in volume). Immediately after extraction, the specimen is placed in a primary fixative such as glutaraldehyde, followed by secondary fixation in osmium tetroxide after 1.5 hours. Fixation is carried out at room Temperature for several hours.

2. Washing the specimen in a buffer solution (pH 7.3–7.4).

3. Dehydration in an ascending series of alcohol concentrations: 50%, 60%, 70%, 80%, 90%, and 100°.

4. Embedding the specimen in specialized synthetic resins (such as Epon and Araldite) capable of polymerization and hardening. To achieve this, the specimen is placed in a capsule filled with liquid epoxy resin and then transferred to a thermostat where polymerization takes place at 58 °С.

5. Sectioning using ultramicrotomes.

6. Staining (contrasting) of the sections using heavy metal salts (lead, tungsten, uranyl acetate). These metal salts precipitate within Cellular Structures and scatter the electrons passing through the specimen.



Last update: 10/08/2026

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