Cell capture system and method of use

• US 10,436,700 B1
• Filed: 07/16/2019
• Issued: 10/08/2019
• 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, Gogoi, Pridyadarshini, Siemer, Christopher, Javdani, Saedeh Sepehri
• Primary Examiner(s): Wecker, Jennifer
• Application Number:
  US16/513,580
• Time in Patent Office: 84 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 system for isolating and analyzing a set of target particles from a biological sample, comprising:

a substrate comprising a set of chambers at a first broad surface, wherein each chamber in the set of chambers is configured to retain a single target particle of the set of target particles and comprises;

a set of walls defining a chamber volume, a chamber length, and a chamber cross-section; and

an open surface permitting access of the single target particle to the chamber volume from a direction perpendicular to the broad surface of the substrate;

an optical element disposed adjacent to the set of chambers, wherein the optical element is configured to direct light transmitted through at least one of the first broad surface and a second broad surface opposing the first broad surface, into the set of chambers;

an inlet channel positioned superior the set of chambers and directly fluidly coupled to the set of chambers, whereby fluid received at the inlet channel is configured to concurrently access each of the chambers in the set of chambers at the open surface of each chamber, wherein fluid received at the inlet channel enters each chamber only by way of the open surface of each chamber; and

a fluid network fluidly coupled to the inlet channel, wherein the fluid network is configured to distribute a reagent fluid in a direction substantially parallel to the broad surface through the inlet channel, permitting access of the reagent fluid to the set of chambers.

2. The system of claim 1, wherein the substrate is optically transparent, and wherein each chamber of the set of chambers is individually optically accessible from a direction perpendicular to the broad surface of the substrate.

3. The system of claim 1, wherein the set of chambers comprises more than 200,000 chambers.

4. The system of claim 1, wherein the set of target particles comprise a set of cells co-captured with a set of functionalized microspheres, as a set of complexes.

5. The system of claim 4, wherein the set of functionalized microspheres comprises a set of magnetic microspheres.

6. The system of claim 1, wherein the optical element comprises at least one of a light reflector, a microlens, a light collimator, a filter, and an diffuser.

7. The system of claim 1, wherein the reagent fluid distributed by the fluid network is configured for processing the biological sample for at least one of:

a single cell proteomic analysis, a nucleic acid analysis, and a genomic sequencing process.

8. The system of claim 1, wherein the chamber length of each chamber prevents egress of the single target particle due to fluid crossflow at the intersection between the inlet channel and the open surface of the chamber.

9. The system of claim 2, further comprising an imaging subsystem positioned at a first side of the substrate and oriented in a direction perpendicular to the broad surface of the substrate.

10. The system of claim 3, further comprising a stage configured to align a selected chamber of the of the set of chambers with an objective of the imaging subsystem.

11. The system of claim 1, wherein each chamber of the set of chambers is fluidly isolated from an adjacent chamber by the set of walls of each chamber.

12. The system of claim 1, wherein the chamber cross-section of each chamber in the set of chambers is arranged parallel to the broad face of the substrate, and wherein the cross-section of each chamber in the set of chambers defines a polygon.

13. The system of claim 1, further comprising a reagent cartridge comprising a set of compartments, wherein each of the set of compartments contains one of a set of reagent fluids, wherein each compartment of the reagent cartridge stores a different reagent fluid.

14. The system of claim 1, further comprising a pumping subsystem system configured to couple to the inlet channel, wherein the pumping subsystem provides a pumping pressure at the inlet channel to promote a controlled flow rate of the reagent fluid through the inlet channel.

15. A system for isolating and analyzing a set of target particles from a biological sample, comprising:

a substrate comprising a set of chambers and a broad face, wherein each chamber in the set of chambers is configured to retain a single target particle of the set of target particles and comprises;

a set of walls defining a chamber volume and a chamber cross-section; and

an open surface permitting access of the single target particle to the chamber volume from a direction perpendicular to the broad surface of the substrate;

an optical element disposed adjacent to the set of chambers of the substrate, wherein the optical element is configured to direct light into the set of chambers;

an inlet channel positioned superior the set of chambers and directly fluidly coupled to the set of chambers, whereby fluid received at the inlet channel is configured to concurrently access each of the chambers in the set of chambers at the open surface of each chamber, wherein fluid received at the inlet channel enters each chamber only by way of the open surface of each chamber; and

a fluid network comprising a set of branches fluidly coupled to the inlet channel, wherein the fluid network is configured to distribute a reagent fluid in a direction substantially parallel to the broad surface through the inlet channel, permitting access of the reagent fluid to the set of chambers.

16. The system of claim 15, wherein the set of target particles comprise a set of cells co-captured with a set of functionalized microspheres, as a set of complexes, and wherein wherein the set of functionalized microspheres comprises a set of magnetic microspheres.

17. The system of claim 16, wherein the chamber volume of each chamber prevents egress of the single target particle due to fluid crossflow at the intersection between the inlet channel and the open surface of the chamber.

18. The system of claim 17, wherein the chamber cross-section of each chamber in the set of chambers is arranged parallel to the broad face of the substrate, and wherein the cross-section of each chamber in the set of chambers defines a polygon.

19. The system of claim 15, wherein each chamber of the set of chambers is fluidly isolated from an adjacent chamber by the set of walls of each chamber.

20. The system of claim 15, wherein the reagent fluid distributed by the fluid network is configured for processing the biological sample for at least one of:

a single cell proteomic analysis, a nucleic acid analysis, and a genomic sequencing process.