Cell capture system and method of use

• US 10,416,070 B1
• Filed: 05/30/2019
• Issued: 09/17/2019
• 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:
  US16/426,322
• Publication Number:
  US 20190293546A1
• Time in Patent Office: 110 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 15/1463 : using image analysis for extracting features of the particle
• CPC Class Codes: G01N 15/1475 : using image analysis for extracting features of the particle
• 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 for automatically processing target particles in single-particle format, the method comprising:

at a workstation, receiving a substrate comprising a network having an input channel at an upstream end, one or more output channels at a downstream end, and a set of structures in communication with the input channel and the one or more output channels, wherein flow from the input channel is configured to reach at least one of the one or more output channels only by way of the set of structures;

at the workstation, retaining a container comprising a set of target particles;

from a controller in communication with a handling subsystem of the workstation, receiving a set of instructions for automatic processing of the set of target particles, in single-particle format; and

based upon the set of instructions;

with the handling subsystem, transmitting the set of target particles into the network;

isolating each of the set of target particles in single-particle format with an isolation material, within the network of the substrate, and

delivering, by way of the handling subsystem, a reagent material from a reagent container into the input channel of the substrate.

2. The method of claim 1, wherein isolating each of the set of target particles in single-particle format comprises isolating a set of target cells of a sample by way of a set of structures of the network.

3. The method of claim 2, wherein isolating each of the set of target cells comprises separating each target cell from adjacent target cells with a set of barriers configured to fluidly isolate each target cell from adjacent target cells within the network, wherein the set of barriers block fluid flow between adjacent target cells.

4. The method of claim 3, wherein isolating comprises segregating each of the set of target cells in single-cell format in a spatially arranged array within the network.

5. The method of claim 4, wherein the network comprises 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:

a chamber having an upstream end, and a pore channel, fluidly coupled to the chamber at a downstream end of the chamber, having a pore channel width configured to block egress of a particle,

wherein the input channel is directly fluidly coupled to the upstream end of each chamber of the plurality of pores,

wherein the output channel is directly fluidly coupled to each pore channel of the plurality of pores,

wherein flow from the input channel is configured to reach the output channel only by way of the plurality of parallel pores, and

wherein isolating each of the set of target cells in single-cell format comprises isolating the set of target cells in single-cell format at the plurality of pores.

6. The method of claim 1, wherein the reagent material is configured for performing at least one of:

a molecular reaction, immunostaining, a single cell proteomic analysis, nucleic acid analysis, and genomic sequencing individually for each target particle of the set of target particles.

7. The method of claim 1, wherein the reagent material is configured for performing a polymerase chain reaction (PCR) process, and wherein the method further comprises performing the PCR process individually for each of the set of target particle in single-particle format.

8. The method of claim 1, further comprising receiving instructions for performing at least one of:

pipetting, aliquoting, mixing, pumping, and monitoring fluid delivery; and

executing the set of instructions with the handling subsystem.

9. The method of claim 8, wherein the set of target particles comprises a set of target cells, the method further comprising:

automatically transmitting a subportion of the set of target cells from the network in a viable state, with the handling subsystem.

10. The method of claim 1, wherein transmitting the sample with the set of target particles into the network comprises transmitting a cell suspension comprising a set of target cells into the network.

11. The method of claim 1, wherein isolating with the isolation material comprises delivering the isolation material, comprising at least one of an oil and a hydrogel, into an isolation inlet of the substrate in communication with the network.

12. The method of claim 1, further comprising co-capturing a set of target cells with a set of functionalized magnetic particles as the set of target particles, thereby forming a set of complexes, wherein isolating the set of target particles in single-particle format comprises isolating the set of complexes in single-complex format.

13. The method of claim 12, further comprising magnetically separating the set of target cells from the sample, by way of the set of functionalized magnetic particles.

14. A method for automatically processing target particles in single-particle format, the method comprising:

at a workstation, receiving a substrate comprising a network having an input channel at an upstream end, an output channel at a downstream end, and a set of structures in communication with the input channel and the output channel;

at the workstation, retaining a sample comprising a set of target cells;

at the workstation, retaining a volume comprising a set of functionalized particles;

from a controller in communication with a handling subsystem of the workstation, receiving a set of instructions for automatic processing of the set of target cells, in single cell format; and

based upon the set of instructions;

with the handling subsystem, transmitting the sample and the volume into the network;

co-capturing the set of target cells with the set of functionalized particles within the network in single-particle format;

isolating the set of target cells with an isolation material, within the network, and

delivering, by way of the handling subsystem, a reagent material from a reagent container into the input channel of the substrate.

15. The method of claim 14, wherein the set of functionalized particles comprise functionalized magnetic particles configured to couple with the set of target cells.

16. The method of claim 15, further comprising magnetically separating the set of target cells from the sample, by way of the set of functionalized magnetic particles.

17. The method of claim 16, further comprising isolating the set of target cells and the set of functionalized particles as a set of complexes within the network, in single-complex format.

18. The method of claim 17, further comprising:

based on the set of instructions automatically delivering the isolation material, comprising an oil, into an isolation inlet of the substrate, and automatically isolating the set of complexes in single-complex format, as a spaced array, with the oil.

19. The method of claim 18, further comprising automatically transmitting a subportion of the set of target cells from the network in a viable state, with the handling subsystem.

20. The method of claim 19, wherein the reagent material is configured for performing a polymerase chain reaction (PCR) process, and wherein the method further comprises, based upon the set of instructions, performing the PCR process individually for each of the set of target cells in single-cell format.