METHOD FOR MOLECULAR QUANTITATIVE DETECTION OF LOCAL PREVALENCE OF NON-SMALL CELL LUNG CANCER BY DOUBLE IMMUNOFLUORESCENT STAINING OF NORMAL AND TUMOUR TISSUE

摘要

FIELD: medicine.;SUBSTANCE: invention relates to medicine. Method of immunofluorescence detection of local prevalence of tumour process in non-small cell lung cancer (NSCLC) involves preparing a unicellular suspension from a tissue sample of NSCLC, fixation in 4 % formalin solution, washing with phosphate buffer pH = 7.4, incubation with DNA dye Hoechst 33258, double washing from antibodies with 0.05 % BSA in phosphate buffer pH = 7.4, conducting fluorescence analysis on flow cytofluorimeter, wherein the incubation is carried out simultaneously with two types of primary antibodies - rabbit antibodies to TUBB3 and mouse antibodies to cytokeratins, wherein the primary antibodies to TUBB3 and cytokeratin are added simultaneously and incubated overnight at a temperature of +4 °C, and simultaneously with two types of secondary antibodies - anti-rabbit antibodies conjugated with DyLight 650 dye, and anti-mouse antibodies conjugated with DyLight 488, plotted histograms, TUBB3 expression levels are compared in epithelial cells of tumour and surrounding normal tissue, where increased expression of TUBB3 characterizes local prevalence of tumour process in NSCLC.;EFFECT: invention enables to quantitatively evaluate TUBB3 protein expression in epithelial cells of tumour tissue.;1 cl, 2 dwg