Cell capture system and method of use

• US 10,345,219 B2
• Filed: 07/24/2017
• Issued: 07/09/2019
• 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): Wecker, Jennifer
• Application Number:
  US15/657,553
• Publication Number:
  US 20170322142A1
• Time in Patent Office: 715 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 capturing particles in single-particle format, comprising:

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, and having a pore channel width configured to block particle egress, wherein adjacent chambers of the plurality of pores are separated by at least one barrier configured to block fluid flow in a direction parallel to fluid flow through their pore channels;

an inlet channel directly fluidly coupled to the upstream end of each chamber of the plurality of pores; and

an outlet channel directly fluidly coupled to each pore channel of the plurality of pores, wherein flow from the inlet channel is configured to reach the outlet channel only by way of the plurality of pores.

2. The system of claim 1, further comprising a second array having a second plurality of pores, a second inlet channel directly fluidly coupled to the second array, and a second outlet channel directly fluidly coupled to the second inlet channel only by way of the second plurality of parallel pores.

3. The system of claim 2, further comprising a feeder channel that couples the inlet channel and the second inlet channel in parallel.

4. The system of claim 1, wherein fluid coupling between the inlet channel and the outlet channel of the array, only by way of the plurality of pores, provides a volumetric fluid flow rate within inlet channel that is substantially equal to the volumetric fluid flow rate within the outlet channel.

5. The system of claim 1, wherein each of the plurality of pores has a non-constant cross section along its flow axis.

6. The system of claim 1, wherein the pore channel widths of successive pores of the plurality of pores decrease along a flow direction through the inlet channel.

7. The system of claim 1, wherein the inlet channel is partially defined by a self-sealing wall at least at one end of the inlet channel.

8. The system of claim 7, further including a particle removal tool operable to penetrate the self-sealing wall and to retrieve a captured particle from one of the plurality of pores.

9. The system of claim 1, further comprising a capillary particle removal unit operable to retrieve a captured particle from one of the plurality of pores.

10. The system of claim 1, further including:

an inlet manifold, defined on the broad face of the substrate, fluidly coupled to the inlet channel of the array; and

an outlet manifold, defined on the broad face of the substrate, fluidly coupled to the outlet channel of the array.

11. A method for capturing particles, comprising:

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;

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 adjacent chambers of the plurality of pores are separated by at least one barrier configured to substantially block fluid flow in a direction parallel to fluid flow through the pore channels;

an inlet channel directly fluidly coupled to the upstream end of each chamber of the plurality of pores; and

an outlet channel directly fluidly coupled to each pore channel of the plurality of pores, wherein flow from the inlet channel is configured to reach the outlet channel only by way of the plurality of parallel pores; and

delivering a sample of particles through the inlet channel, into the plurality of pores, and allowing fluid flow from the plurality of pores through the outlet channel, thereby capturing particles in single-particle format.

12. The method of claim 11, wherein the particles comprise circulating tumor cells (CTCs).

13. The method of claim 11, wherein the particles comprise circulating stem cells (CSCs).

14. The method of claim 11, wherein providing the plurality of pores comprises providing the plurality of pores at a broad face of a substrate.

15. The method of claim 14, further comprising receiving a particle removal tool into at least one of the plurality of pores, the cell removal tool providing a capillary effect that draws at least one particle into the particle removal tool.

16. The method of claim 11, further comprising backflowing fluid through the outlet channel toward the inlet channel, thereby releasing captured particles.

17. The method of claim 11, further comprising providing a second array having a second plurality of pores, a second inlet channel directly fluidly coupled to the second array, and a second outlet channel directly fluidly coupled to the second inlet channel only by way of the second plurality of parallel pores.

18. The method of claim 17, further comprising providing a feeder channel directly coupled to the inlet channel and the second inlet channel, and delivering a volume of particles into the feeder channel, thereby delivering particles into the inlet channel and the second inlet channel in parallel.

19. The method of claim 11, further comprising introducing an isolating material in the plurality of pores, thereby trapping captured particles at the plurality of pores.

20. The method of claim 19, further including transitioning the isolating material from a flow state to a set state and processing captured particles at the plurality of pores with a process reagent upon introducing the process reagent into the plurality of pores.