Cell capture system and method of use

• US 10,401,277 B2
• Filed: 07/27/2018
• Issued: 09/03/2019
• 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, Priyadarshini, Siemer, Christopher, Javdani, Saedeh Sepehri
• Primary Examiner(s): Hixson, Christopher Adam
• Application Number:
  US16/048,104
• Publication Number:
  US 20180364148A1
• Time in Patent Office: 403 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 for isolating and analyzing a set of cells from a biological sample, comprising:

a substrate defining a set of chambers at a broad surface, wherein each chamber in the set of chambers is configured to retain a single cell of the set of cells 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 cell to the chamber volume from a direction perpendicular to the broad surface of the substrate;

a light reflector disposed within the substrate between the broad surface and a second broad surface and separated from a chamber in the set of chambers by a thickness of the substrate defining a wall of the chamber, wherein the light reflector is configured to reflect light incident from a direction perpendicular to the broad surface into the chamber through the wall;

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 can concurrently access each of the chambers in the set of chambers at the open surface of each chamber;

a fluidic manifold fluidly coupled to the inlet channel, wherein the fluidic manifold is operable 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;

an imager 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 of the set of chambers with an objective of the imager.

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 substrate includes a set of optical elements defined within the substrate and detectable by the imager, wherein the set of optical elements is configured to register the location of the selected chamber relative to the substrate.

4. The system of claim 1, further comprising a cell removal tool, configured to operate in coordination with the imager to remove a selected cell from the selected chamber.

5. The system of claim 4, wherein the cell removal tool comprises a hollow needle configured to form a substantially fluidly isolated volume within the selected chamber.

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

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

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

9. The system of claim 1, wherein each 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 egress of the single cell from the chamber due to fluid flow through the chamber volume of the chamber.

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

11. The system of claim 1, wherein the set of chambers comprises 1,000,000 individual chambers.

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 12, wherein the cross-section of each chamber in the set of chambers defines a rectangle.

14. The system of claim 1, further comprising a plurality of compartments fluidly connected to the inlet channel, wherein each of the plurality of compartments stores one of a plurality of reagent fluids, wherein each compartment of the cartridge stores a different reagent fluid.

15. The system of claim 14, wherein the plurality of reagent fluids stored in the plurality of compartments comprises reagent fluids permitting a plurality of biochemical processes to be performed within the set of chambers.

16. The system of claim 15, wherein the plurality of biochemical processes comprises one of immunostaining, single cell proteomic analysis, nucleic acid analysis, and genomic sequencing of the set of cells retained within the set of chambers.

17. The system of claim 1, further comprising a pumping system coupled to an inlet and an outlet of the inlet channel, wherein the pumping system provides a pumping pressure at one of the inlet and the outlet of the inlet channel to promote a controlled flow rate of the reagent fluid through the inlet channel.

18. The system of claim 17, wherein the pumping pressure provided by the pumping system is less than 10,000 pascals.

19. The system of claim 1, further comprising a thermal control system comprising a heater physically coupled to a second broad surface of the substrate directly opposing the broad surface of the substrate, wherein the thermal control system is operable to adjust an output of the heater and thereby the temperature of the set of chambers according to a preset temperature profile.