METHOD OF IMMUNOHISTOCHEMICAL ANTIGENS DETECTION IN PREPARATIONS OF PRODUCTIVE AND NON-PRODUCTIVE ANIMALS ORGANS OF LONG-TERM STORAGE IN FIXTURES Russian patent published in 2017 - IPC G01N33/53 

FIELD: medicine.

SUBSTANCE: for immunohistochemical detection of antigens in preparations of productive and unproductive animals organs of long-term storage in fixators, the surface of a slide is treated to glue the sections with an adhesive based on egg protein and glycerin at a 2:1 ratio. Then, sections dewaxing and rehydration are carried out. In this case, after dewaxing and rehydration, the sections are lowered into a clean container with distilled water for 5 minutes. Then the slices are placed in H₂O₂ for 10-12 minutes, after operation of a steamer with the citrate buffer for 40-45 minutes. At that, the slides are not removed from the steamer within 20 minutes after termination of its operation. The sections are rinsed with distilled water, then placed in a Twin 20x buffer. To form a dry field, slices are dabbed with disposable paper towels and a hydrophobic layer is made around the slices with a marker. Then, a blocking solution is applied, which is 5% bovine serum albumin, for 10-12 minutes. Primary antibodies are applied and placed in a thermostat at 27°C for 24-26 hours in a "wet chamber", after 14-15 hours, the primary antibodies are washed away with Tween 20x buffer for 5-7 minutes. At that, the sections are dipped and the secondary antibodies are applied to them. They are put in the thermostat for 1-2 hours in a "wet chamber" at a temperature of 27°C. The secondary antibodies are washed away. Chromogen on the bebiothin detection system is used. Stained for 3 minutes, the chromogen is washed away in three portions of distilled water for 5-7 minutes. Slides are stained with Mayer hematoxylin for 5-7 minutes. Then the slides are rinsed in distilled water and lowered into alkaline water for 10-15 seconds.

EFFECT: invention allows to identify antigens in histological preparations, after their long storage in fixators.

8 dwg, 4 ex