Methods, systems and devices for multiple single-cell capturing and processing using microfluidics

A method for multiple single-cell capturing and processing using microfluidics, the method comprising:

• a) loading a plurality of cells into a microfluidic device;
• b) flowing the plurality of cells to a first capture configuration of the microfluidic device;
• c) capturing a first single cell from the plurality of cells in the first capture configuration, wherein the first capture configuration comprises one or more physical barriers configured to capture a single cell from the plurality of cells and wherein the first capture configuration is coupled to a first multi-chamber reaction configuration;
• d) flowing a first remaining plurality of cells from the plurality of cells through one or more first bypass channels to a second capture configuration wherein the second capture configuration comprises one or more physical barriers configured to capture a single cell from the plurality of cells and wherein the second capture configuration coupled to a second multi-chamber reaction configuration;
• e) capturing a second single cell from the first remaining plurality of cells in the second capture configuration;
• f) contacting the first and second single cells with a lysis reagent to release one or more cell constituents of each respective captured cell;
• g) flowing the one or more constituents of the first captured cell into the first multi-chamber reaction configuration and flowing the one or more constituents of the second captured cell into the second multi-chamber reaction configuration;
• and
• h) processing the one or more constituents of the first captured single cell in the first multi-chamber reaction configuration and processing the one or more constituents of the second captured single cell in the second multi-chamber reaction configuration to produce harvest products with respect to at least the first captured single cell and the second captured single cell,
• wherein the multi-chamber reaction configurations each comprise:
• a first reaction chamber coupled to the capture configuration and a second reaction chamber coupled to the first reaction chamber, wherein the capture configuration, the first reaction chamber, and the second reaction chamber are arranged so that fluid transported from the capture configuration to the second reaction chamber must pass through the first reaction chamber, and wherein the first and second reaction chambers are coupled though two non-overlapping fluid flow paths,
• valves positioned to fluidically isolate the first and second reaction chambers from each other when closed, and,
• a valve positioned to fluidically isolate the capture configuration from the multi-chamber reaction configuration when closed,
• wherein steps (a)-(h) are carried out in the microfluidic device.