METHOD FOR DETECTION OF ONCOLOGICAL MARKERS IN TUMOR TISSUE SAMPLES AND DEVICE FOR ITS IMPLEMENTATION Russian patent published in 2024 - IPC G01N1/30 G01N21/01 G01N21/05 G01N21/64 

FIELD: medicine.

SUBSTANCE: group of inventions relates to a method for detecting oncological markers in tumor tissue samples, which includes preliminary placement of a section of a tumor tissue sample with thickness of 20–200 mcm on a glass, fixation and permeabilization of the section of the tumor tissue sample, then the glass with the tumor tissue sample is washed and placed on the flow cell, after which sealing is carried out with the possibility of forming a closed volume of a flow chamber by the glass surfaces and the flow cell with the tumor tissue sample enclosed therein, wherein the height of the flow chamber formed by the surfaces of the glass and the flow cell should be 2–5 times greater than the thickness of the analyzed tissue section, then, through the inlet opening, a buffer is supplied to block non-specific binding, in a volume equal to 3–5 volumes of the flow chamber, after which a washing buffer is supplied into the inlet opening in a volume equal to 5–10 volumes of the flow chamber, further, a solution containing a mixture of two or more different populations of spatially oriented conjugates of single-domain antibodies and fluorescent nanocrystals is fed into the inlet opening, in a volume equal to 3–5 volumes of the flow chamber, after which a washing buffer is supplied to the inlet opening in a volume equal to 5–10 volumes of the flow chamber, wherein the supply of liquids through the inlet port of the flow cell is carried out using a pump, and the pressure of the liquid in the flow chamber is controlled by varying the flow through the outlet port of the flow cell, finally, nanocrystals fluorescence is detected in specified optical ranges using a confocal fluorescent microscope, and also relates to a device for implementing the method, consisting of a flow cell, an inlet port, an outlet port, a drain tube and an outlet flow rate control module, wherein the flow cell is a flat plate made of an optically transparent polymer having a through inlet opening, perpendicular to the plane of the plate, connected to the input channel of the input microfluidic module, output channels of which are connected to the end of the flow chamber, the opposite end of the flow chamber is connected to the inlet channels of the output microfluidic module, outlet channel of which is connected to a through outlet opening located perpendicular to the plane of the plate, to the inlet opening of the flow cell, an inlet port is connected for connecting a pump and supplying liquid, and to the outlet opening of the flow cell, an outlet port is connected by a drain tube to the liquid outlet flow rate control module, wherein the inlet and outlet ports are connected to the flow cell on the side of the plane of the opposite surface containing the flow chamber.

EFFECT: group of inventions provides higher efficiency of immunohistochemical studies due to possibility of detection of oncological markers in thick sections of tumor tissue with thickness of up to 200 mcm and simultaneous immunohistochemical volumetric staining of two or more oncological markers in flow staining mode.

16 cl, 1 dwg, 2 ex