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, and wherein the set of structures comprises a set of microfluidic segments coupled to the inlet channel and the one or more outlet channels;
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, 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 microfluidic segments comprises one or more isolation material containing segments configured to separate the set of target cells from non-target material of the fluid sample.
3. The method of claim 1 , wherein the set of structures comprise a set of branched microfluidic channels.
4. The method of claim 3 , wherein one or more of the set of branched microfluidic channels terminate at the one or more outlet channels.
5. The method of claim 3 , wherein the set of branched microfluidic channels comprises at least one channel with a characteristic dimension of less than 100 micrometers.
6. The method of claim 1 , wherein receiving the fluid sample comprises receiving a set of viable target cells into the fluidic network at a flow rate for maintenance of cell viability.
7. The method of claim 1 , wherein transmitting material associated with the set of target cells from the fluidic network comprises transmitting the set of target cells, in a viable state, from the fluidic network.
8. The method of claim 1 , wherein transmitting material comprises transmitting material associated with the set of target cells into a retrieval channel of the fluidic network.
9. The method of claim 1 , wherein capturing and partitioning comprises co-capturing the set of target cells with a set of functionalized microparticles within the fluidic network in an arrayed distribution as a set of complexes.
10. The method of claim 1 , wherein capturing and partitioning comprises isolating each of the set of complexes in single-complex format within the fluidic network with the isolation material.
11. The method of claim 1 , wherein the isolation material comprises an oil.
12. The method of claim 1 , further comprising contacting the set of target cells with a lysis reagent, thereby generating cellular lysate from the set of target cells.
13. The method of claim 12 , further comprising performing single-cell analyses for individual cells of the set of target cells with the cellular lysate.
14. The method of claim 13 , wherein performing single-cell analyses comprises performing a downstream processing operation with the cellular lysate, 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.
15. The method of claim 1 , wherein receiving the fluid sample comprises receiving 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.
16. 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 comprising a collection volume, and delivering the material into the collection volume.
17. The method of claim 1 , wherein providing the fluidic network comprises:
providing a set of cell-isolators each cell-isolator in the set of cell-isolators displaced from adjacent cell-isolators and comprising: an upstream end and a downstream end configured to isolate the set of target cells in single-cell format, wherein a cell-isolator of the set of cell-isolators comprises a region of microfluidic structure and a portion of isolation material,
wherein the inlet channel is fluidly coupled to the upstream end of at least one of the set of cell-isolators.
18. The method of claim 1 , further comprising performing a molecular reaction with material associated with the set of target cells.
19. The method of claim 18 , further comprising transmitting the set of target cells from the fluidic network prior to performing the molecular reaction.
20. The method of claim 18 , wherein the molecular reaction is associated with a single-cell expression analysis.