METHOD OF ANTIGEN RETRIEVAL DURING IMMUNOCYTOCHEMICAL REACTIONS Russian patent published in 2020 - IPC G01N1/30 

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to diagnostic histologic examinations. Disclosed is a method of antigens retrieval in immunocytochemical reactions involving fixing preparation in 10 % formalin for 24 hours at room temperature, procedure for dehydration, paraffin preparation, preparation of histological sections with thickness of 5 to 10 mcm, application thereof on adhesive slides, dewaxing in xylene, conducting on alcohols of lower concentration, washing for 5 minutes in distilled water, characterized by that after washing sections for 22 minutes subjected to thermal retrieval in 10 % aqueous solution of sodium thiosulphate, preheated to 45–60 °C, cooled glasses are washed with distilled water, left in 0.01 M phosphate-salt buffer with pH 7.4 for 5 minutes at room temperature, then blocking solution is applied for 10 minutes at room temperature, excess solution is drained and, without washing preparations, first antibodies are applied on glass, after which the preparations are placed in a moist chamber and incubated in a thermostat at temperature of 27 °C for 20 hours, then washed in three changes of distilled water for 5 minutes in each, after which secondary antibodies are applied and incubated in a moist chamber in a thermostat at temperature of 27 °C for 30 minutes, are washed in three changes of distilled water for 5 minutes in each, then chromogen solution is applied and during 1–3 minutes formation of colored product of histochemical reaction is monitored under a microscope, chromogen is washed off and preparations are washed in 2-3 portions of distilled water for 5 minutes in each, if necessary, the preparations are colored by histological dyes, dehydrated in alcohols of the ascending fortress, clarified in xylene and enclosed in any non-water-soluble confinement medium.

EFFECT: high efficiency and industrial safety of the method of antigens retrieval.

1 cl, 2 dwg, 1 ex