Cell capture system and method of use

• US 10,533,936 B1
• Filed: 10/11/2019
• Issued: 01/14/2020
• Est. Priority Date: 08/01/2011
• Status: Active Grant

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Celsee Diagnostics, Inc. (Bio-Rad Laboratories Incorporated)
• Inventors: Handique, Kalyan
• Primary Examiner(s): Wecker, Jennifer
• Application Number:
  US16/599,704
• Publication Number:
  US 20200041402A1
• Time in Patent Office: 95 Days
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0672 Integrated piercing tool
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0848 Specific forms of parts of containers
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/168 Specific optical properties, e.g. reflective coatings
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/021 Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502746 characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: G01N 1/20 for flowing or falling materials G01N1/2035 G01N1/12, G01N1/14 take precedence
• CPC Class Codes: G01N 1/28 Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Qmounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 ; laboratory gas handling apparatus B01L5/00
• CPC Class Codes: G01N 1/40 Concentrating samples
• CPC Class Codes: G01N 1/405 by adsorption or absorption
• CPC Class Codes: G01N 1/4077 : by other techniques involving separation of suspended solids
• CPC Class Codes: G01N 15/01 : specially adapted for biological cells, e.g. blood cells investigating sedimentation of particle suspensions in blood G01N15/05
• CPC Class Codes: G01N 15/0272 : with screening; with classification by filtering B01D takes precedence
• CPC Class Codes: G01N 15/1433 : using image recognition
• CPC Class Codes: G01N 15/1436 : the optical arrangement forming an integrated apparatus with the sample container, e.g. a flow cell
• CPC Class Codes: G01N 15/1484 : microstructural devices
• CPC Class Codes: G01N 15/149 : specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/1006 : for cytology
• CPC Class Codes: G01N 2015/1486 : Counting the particles
• CPC Class Codes: G01N 2015/1497 : Particle shape
• CPC Class Codes: G01N 2035/00158 : Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 33/48728 : Investigating individual cells, e.g. by patch clamp, voltage clamp investigating individual particles in general G01N15/10
• CPC Class Codes: G06V 20/69 : Microscopic objects, e.g. biological cells or cellular parts

Abstract

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.

Claims

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 viable target cells into the fluidic network at a flow rate for maintenance of cell viability;

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, further comprising receiving a set of microparticles into the fluidic network.

3. The method of claim 2, wherein capturing and partitioning comprises co-capturing the set of viable target cells with the set of microparticles within the fluidic network in an arrayed distribution.

4. The method of claim 3, wherein capturing and partitioning comprises isolating each of the set of complexes in single-complex format within the fluidic network with the isolation material.

5. The method of claim 1, wherein the isolation material comprises an oil.

6. The method of claim 1, further comprising contacting the set of viable target cells with a lysis reagent, thereby generating cellular lysate from the set of viable target cells.

7. The method of claim 6, further comprising performing performing single cell-analyses for individual cells of the set of target viable cells with the cellular lysate.

8. The method of claim 6, further comprising performing a downstream processing operation with the cellular lysate derived from the set of viable target cells, 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 the fluidic network comprises at least one channel with a characteristic dimension of less than 100 micrometers.

10. The method of claim 1, wherein receiving the fluid sample at the flow rate 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.

11. 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.

12. 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 viable target cells into the fluidic network at a flow rate for maintenance of cell viability;

receiving a set of microparticles into the fluidic network;

capturing and partitioning the set of viable cells with the set of functionalized microparticles, wherein capturing and partitioning comprises isolating each of the set of viable target cells in single-cell format with one or more of the set of microparticles, with an isolation material within the microfluidic network;

receiving a lysis reagent into the fluidic network, thereby generating cellular lysate from the set of viable cells; and

transmitting material derived from the set of viable target cells from the fluidic network.

13. The method of claim 12, further comprising performing a molecular reaction with the cellular lysate, thereby performing single cell-analyses for individual cells of the set of target viable cells.

14. The method of claim 13, further comprising transmitting the cellular lysate from the fluidic network prior to performing the molecular reaction.

15. The method of claim 12, wherein capturing and partitioning comprises segregating each of the set of viable target cells in single-cell format, with one or more of the set of microparticles, in an arrayed distribution within the fluidic network.

16. The method of claim 12, wherein receiving the fluid sample with the set of viable target cells comprises receiving a cell suspension comprising a dilution of viable cells.

17. The method of claim 12, wherein receiving the fluid sample at the flow rate 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.

18. The method of claim 12, 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.

19. The method of claim 12, 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, transmitting material into a collection volume, and delivering the material into the collection volume.

20. The method of claim 19, 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.