METHOD OF PREPARATION AND CYTOMIC ANALYSIS OF HEPATOCYTES OF LABORATORY ANIMALS AND HUMANS FOR ASSESSING CYTOGENETIC AND CYTOTOXIC ACTION OF FACTORS OF DIFFERENT NATURE Russian patent published in 2020 - IPC G01N33/483 

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine, namely cytology, and can be used for assessing cytogenetic and cytotoxic action of various factors on hepatocytes of experimental animals and humans. That is ensured by producing liver hepatocytes after animal euthanasia or human biopsy. Preparation is then fixed in 10 % formalin and alkaline dissociation of 50 % KOH is carried out at 25–37 °C for three hours. Thereafter, cell suspension is applied by smear method on dry slide glass, staining with aceto-orsine with green-green docking of cytoplasm and analyzed by 1000 hepatocytes light microscope. Hepatocyte cytomic analysis involves determining the following cytological parameters: determining cells with microcapsules, cells with nuclear protrusions, cells with internuclear bridges and the sum of cells with these parameters. Additionally quantitative change of single-core diploid (2n), tetraploid (4n), polyploid (≥8n), binuclear diploid (2n + 2n), tetraploid (4n + 4n) and polyploid (≥8n + 8n) hepatocytes. Ploidy index is then calculated as the ratio of the sum of polyploid hepatocytes to the sum of diploid hepatocytes and the nuclear index as the ratio of the sum of binuclear hepatocytes to the sum of mononuclear hepatocytes.

EFFECT: group of inventions provides simplifying the method of preparing hepatocytes for analysis, as well as high information value of assessing cytogenetic and cytotoxic action of various factors on the body.

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