A method for capturing a biomolecule in each single cell from a sample comprising a plurality of cells, comprising:
- providing a system for capturing the biomolecule, the system comprising:
- a common inlet for serially introducing the sample, a washing buffer, and a cell lysis reagent into a flow channel;
- a two-dimensional array having a plurality of single cell capture holes formed on a surface of a substrate, and biomolecule capture areas inside of the substrate each comprising a biomolecule capture member for capturing the biomolecule extracted from individual cells which are respectively captured by the plurality of single cell capture holes;
- the flow channel for flowing the sample, which comprises the plurality of cells to be assayed, from the common inlet and in a first direction in parallel with the surface of the substrate as a first flow;
- a structure provided on the surface of the substrate so as to be opposed to the first direction on a downstream side of each of the plurality of single cell capture holes;
- a first syringe connected via a first discharging tube to a first outlet;
- a first mass flow controller capable of measuring and controlling a first flow volume of the first discharging tube, wherein the first mass flow controller controls a first flow rate of the first flow from the common inlet to the first outlet;
- a second syringe connected via a second discharging tube to a second outlet;
- a second mass flow controller capable of measuring and controlling a second flow volume of the second discharging tube, wherein the second mass flow controller controls a second flow rate of a second flow from the common inlet to the second outlet such that the first flow rate of the first flow is greater than that of the second flow rate of the second flow that is orthogonal to the first flow upon cell capture and the second flow rate of the second flow is greater than the first flow rate of the first flow upon biomolecule capture;
- a reagent dispensing unit comprising a pressurizer and a set of reagent-accommodating dispensers;
- a first-reagent-accommodating dispenser in the set of reagent-accommodating dispensers containing the sample and dispensing the sample into the common inlet;
- a second-reagent-accommodating dispenser in the set of reagent-accommodating dispensers containing the washing buffer and dispensing the washing buffer into the common inlet;
- a third-reagent-accommodating dispenser in the set of reagent-accommodating dispensers containing the cell lysis reagent and dispensing the cell lysis reagent into the common inlet;
- and
- an automatic moving stage unit for moving a position of the common inlet relative to the reagent dispensing unit, wherein a first position of the common inlet is associated with dispensing the sample, a second position is associated with dispensing the washing buffer, and a third position of the common inlet is associated with dispensing the cell lysis reagent;
- generating the first flow of the sample from the common inlet via the flow channel of the system to the first outlet;
- capturing, based on the first flow of the sample, the individual cells by each of a set of single cell capture holes on the two-dimensional array;
- washing the two-dimensional array by generating a third flow of the washing buffer, in parallel with the surface of the substrate, from the common inlet to the first outlet via the flow channel of the system
- lysing each captured cell by generating the second flow of the cell lysis reagent from the common inlet to the second outlet that is orthogonal to the one surface of the substrate;
- and
- capturing, based on the second flow of the cell lysis reagent, the biomolecule by each of a set of biomolecule capture members of the two-dimensional array.