Cell capture system and method of use

• US 10,408,737 B1
• Filed: 06/17/2019
• Issued: 09/10/2019
• 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:
  US16/443,140
• Publication Number:
  US 20190302001A1
• Time in Patent Office: 85 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 system comprising:

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

a set of walls defining a chamber volume, and

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

wherein the set of walls of each chamber of the set of chambers fluidly isolates adjacent chambers of the set of chambers, and wherein fluid received at the inlet channel enters each chamber only by way of the open surface of each chamber; and

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, and wherein the system is configured to receive a distribution of a reagent fluid in a direction substantially parallel to the broad surface through the inlet channel and to permit access of the reagent fluid to the set of chambers.

2. The system of claim 1, wherein a chamber length of each chamber of the set of chambers is configured to prevent single particle egress due to fluid crossflow at interfaces between the inlet channel and the open surfaces of the set of chambers.

3. The system of claim 1, wherein a chamber of the set of chambers is fluidly connected to a set of fluid channels, wherein the set of fluid channels defines a characteristic channel dimension preventing single particle egress from the chamber due to fluid flow through the chamber volume of the chamber.

4. The system of claim 1, wherein a 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.

5. The system of claim 1, wherein the set of chambers comprises from 10,000 to 1,000,000 individual chambers.

6. The system of claim 1, wherein a characteristic dimension of each chamber is less than 200 micrometers.

7. 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.

8. The system of claim 7, wherein the substrate further comprises a set of optical elements defined within the substrate and configured to register the location of each of the set of chambers relative to the substrate.

9. The system of claim 8, further comprising an imaging subsystem comprising:

an optical assembly positioned at a first side of the substrate and oriented in a direction perpendicular to the broad surface of the substrate; and

an automated stage configured to align a selected chamber of the set of chambers with an objective of the optical assembly.

10. The system of claim 1, further comprising a fluid delivery module configured to fluidly couple to the inlet channel, the fluid delivery module configured to provide the distribution of the reagent fluid in a direction substantially parallel to the broad surface through the inlet channel.

11. The system of claim 10, wherein the fluid delivery module further comprises a pumping system coupled to the inlet channel, wherein, during provision of the distribution of the reagent fluid, the pumping system provides an actuation pressure at the inlet channel to promote a controlled flow rate of the reagent fluid into the inlet channel.

12. The system of claim 10, further comprising a reagent container configured to interface with the fluid delivery module, wherein the reagent container comprises a set of fluidly isolated compartments storing a set of reagent fluids, wherein each compartment of the reagent container stores a different reagent fluid.

13. The system of claim 12, wherein the set of reagent fluids stored in the cartridge of the fluid delivery module comprises reagent fluids permitting a biological assay to be performed within the set of chambers, and wherein the biological assay comprises at least one of:

immunostaining, single cell proteomic analysis, nucleic acid analysis, and genomic sequencing of the set of particles retained within the set of chambers.

14. The system of claim 1, further comprising a thermal control subsystem configured to physically contact a second broad surface of the substrate opposing the broad surface of the substrate, wherein the thermal control subsystem is operable to adjust the temperature of the set of chambers according to a protocol.

15. A system comprising:

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

a set of walls defining a chamber volume and configured to provide physical isolation from adjacent chambers, and

an open surface permitting access to the chamber volume from a direction perpendicular to the broad surface of the substrate,

wherein fluid received at the inlet channel enters each chamber only by way of the open surface of each chamber; and

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 be distributed across each of the chambers in the set of chambers and enter the open surface of each chamber.

16. The system of claim 15, wherein a 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, and wherein the set of chambers comprises at least 10,000 chambers.

17. The system of claim 15, 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.

18. The system of claim 17, further comprising an imaging subsystem comprising:

an optical assembly positioned at a first side of the substrate and oriented in a direction perpendicular to the broad surface of the substrate; and

an automated stage configured to align a selected chamber of the set of chambers with an objective of the optical assembly.

19. The system of claim 15, wherein the substrate further comprises an outlet channel downstream of the set of chambers and configured to transmit fluid from the system.

20. The system of claim 15, further comprising a reagent container comprising a set of fluidly isolated compartments storing a plurality of reagent fluids, wherein the reagent container is configured to deliver individual reagent fluids of the plurality of reagent fluids for transmission into the inlet channel.