Cell capture system and method of use
A cell capture system including an array, an inlet manifold, and an outlet manifold. The array includes a plurality of parallel pores, each pore including a chamber and a pore channel, an inlet channel fluidly connected to the chambers of the pores; an outlet channel fluidly connected to the pore channels of the pores. The inlet manifold is fluidly connected to the inlet channel, and the outlet channel is fluidly connected to the outlet channel. A cell removal tool is also disclosed, wherein the cell removal tool is configured to remove a captured cell from a pore chamber.
1. A method comprising:
providing a fluidic network comprising an inlet channel at an upstream end, one or more outlet channels at a downstream end, and a set of structures in fluid communication with the inlet channel and the one or more outlet channels, wherein flow from the inlet channel is configured to reach at least one of the one or more outlet channels only by way of the set of structures;
receiving a fluid sample comprising a set of target cells into the fluidic network;
capturing and partitioning the set of target cells in single-cell format, by way of the set of structures, wherein capturing and partitioning comprises isolating a target cell of the set of target cells in single-cell format with an isolation material within at least a portion of the fluidic network; and
transmitting material associated with the set of target cells from the fluidic network.
2. The method of claim 1 , wherein the set of target cells is co-captured with a set of functionalized microspheres, thereby forming a set of complexes, and wherein capturing and partitioning the set of target cells in single-cell format comprises capturing and partitioning the set of complexes in single-complex format.
3. The method of claim 1 , wherein isolating each of the set of target cells with the isolation material comprises isolating each of the set of target cells with an oil.
4. The method of claim 1 , wherein receiving the fluid sample comprises receiving a diluted fluid sample comprising the set of target cells.
5. The method of claim 1 , wherein transmitting material associated with the set of target cells from the fluidic network comprises establishing communication between the fluidic network and a removal tool, and delivering the material into the removal tool.
6. The method of claim 5 , wherein the removal tool comprises a tip configured to enter the fluidic network, extract the material from the fluidic network, and be reversibly retracted from the fluidic network with the target material.
7. The method of claim 1 , further comprising contacting the target cell with a reagent, wherein the reagent comprises a lysing reagent.
8. The method of claim 7 , further comprising performing a downstream processing operation with cellular lysate of the target cell, wherein the downstream processing operation comprises at least one of: a molecular reaction, an amplification process, immunostaining, single cell proteomic analysis, nucleic acid analysis, and genomic sequencing.
9. The method of claim 1 , wherein capturing and partitioning comprises segregating each of the set of target cells in single-cell format in an arrayed distribution within the fluidic network.
10. The method of claim 9 , wherein providing the fluidic network comprises providing a plurality of pores, each pore in the plurality of pores displaced from at least one other pore in the plurality of pores and comprising: an upstream opening and a downstream opening, having a pore channel width configured to block target particle egress,
wherein the inlet channel is fluidly coupled to the upstream end and at least one of the one or more outlet channels is directly fluidly coupled to the downstream end of each of the plurality of pores.
11. The method of claim 1 , wherein the fluidic network comprises at least one channel with a characteristic dimension of less than 100 micrometers.
12. The method of claim 1 , wherein receiving the fluid sample comprises transmitting the fluid sample into the inlet channel with a pressure difference of less than 10,000 Pascals between the inlet channel and the one or more outlet channels.
13. A method comprising:
providing a fluidic network comprising an inlet channel, an outlet channel, and a set of structures in fluid communication with the inlet channel and one or more outlet channels, wherein flow from the inlet channel is configured to reach at least one of the one or more outlet channels only by way of the set of structures;
receiving a fluid sample comprising a set of target cells into the fluidic network;
capturing and partitioning the set of target cells in single-cell format, by way of the set of structures, wherein capturing and partitioning comprises isolating a target cell of the set of target cells in single-cell format with an isolation material within at least a portion of the fluidic network;
generating cellular lysate upon lysing the target cell with a lysis reagent; and
performing a reaction with the cellular lysate.
14. The method of claim 13 , wherein the fluid sample comprises a diluted fluid sample comprising the set of target cells.
15. The method of claim 14 , wherein the isolation material comprises an oil.
16. The method of claim 15 , wherein at least one of the set of target cells is co-captured with a functionalized microsphere, thereby forming a complex, and wherein capturing and partitioning the set of target cells in single-cell format comprises capturing the complex in the oil.
17. The method of claim 13 , further comprising transmitting the target cell from the fluidic network prior to performing the reaction.
18. The method of claim 17 , wherein the reaction comprises a molecular reaction.
19. The method of claim 17 , wherein transmitting the target cell comprises establishing communication between the fluidic network and a removal tool, delivering the material into the removal tool, and delivering the target cell from the removal tool into a container.
20. The method of claim 13 , wherein the microfluidic network comprises at least one channel with a characteristic dimension of less than 100 micrometers.