METHOD OF STAINING ANTIGENS BY MEANS OF FLUORESCENT ANTIBODIES ON FRESH-FROZEN SECTIONS OF TUMOUR AND METASTASES Russian patent published in 2020 - IPC G01N33/533 

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to a method of staining antigens by means of fluorescent antibodies on fresh-frozen sections of a tumour and metastases. Method for staining antigens by means of fluorescent antibodies on fresh-frozen sections of tumour and metastases, consisting in using fresh-frozen sections, are transferred onto glasses with an adhesive coating, subjected to incubation in 2 % formalin solution, dried in air at temperature of 20–26 °C for an hour; at the rehydration stage, the glass with fixed sections is placed in a vessel with a phosphate salt buffer PBS for 15 minutes, followed by washing in a salt phosphate buffer twice for 5 minutes; next, at the stage of unmasking, the preparations are incubated alternately in 1 % sodium dodecyl sulphate and in a salt phosphate buffer for 15 minutes, then washed in salt phosphate buffer 3 times for 5 minutes; further, at the blocking step, wet slices are coated with 1 % bovine serum albumin solution, blocked for 30 minutes and washed once in salt phosphate buffer for 5 minutes with addition of detergent; followed by an incubation step with primary antibodies in a humid chamber at temperature of 20–26 °C for 1 hour, then sections are washed in phosphate buffer 3 times for 5 minutes; followed by a stage of incubation with secondary antibodies in a moist chamber for 1 hour, followed by washing 3 times for 5 minutes; then nuclei are coloured: a glass is coated with Prolong DAPI reagent, covered with cover glass and fixed with transparent varnish.

EFFECT: method described above enables accurate prediction of metastasis of breast cancer, thereby improving patient observation tactics and reducing analysis time.

1 cl, 5 tbl