System and method for retrieving and analyzing particles

• US 10,564,090 B2
• Filed: 08/02/2019
• Issued: 02/18/2020
• 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, Payne, Austin, Sharma, Vishal, Gleason, Kyle, Gogoi, Priyadarshini, Ganesan, Karthik, Boniface, Brian, Chow, Will
• Primary Examiner(s): Wecker, Jennifer
• Application Number:
  US16/530,201
• Publication Number:
  US 20190353579A1
• Time in Patent Office: 200 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 15/1433 : using image recognition
• 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 2035/00158 : Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 2035/1039 : Micropipettes, e.g. microcapillary tubes
• CPC Class Codes: G01N 35/00029 : provided with flat sample substrates, e.g. slides G01N35/028 takes precedence
• CPC Class Codes: G01N 35/1011 : Control of the position or alignment of the transfer device
• CPC Class Codes: G06V 10/141 : Control of illumination
• CPC Class Codes: G06V 10/145 : Illumination specially adapted for pattern recognition, e.g. using gratings
• CPC Class Codes: G06V 10/147 : Details of sensors, e.g. sensor lenses fingerprint or palmprint sensors G06V40/13; vascular sensors G06V40/145; eye sensors G06V40/19
• CPC Class Codes: G06V 20/69 : Microscopic objects, e.g. biological cells or cellular parts

Abstract

A system and method for isolating and analyzing single cells, including: a substrate having a broad surface; a set of wells defined at the broad surface of the substrate, and a set of channels, defined by the wall, that fluidly couple each well to at least one adjacent well in the set of wells; and fluid delivery module defining an inlet and comprising a plate, removably coupled to the substrate, the plate defining a recessed region fluidly connected to the inlet and facing the broad surface of the substrate, the fluid delivery module comprising a cell capture mode.

Claims

1. A system for cell retrieval from a capture substrate comprising a set of wells distributed across a broad face of the cell capture substrate, the system comprising:

an imaging subsystem configured to sample an image data dataset of one or more of the set of wells;

a retriever subsystem comprising an extractor comprising an extractor tip, an outlet, and a void defined between the extractor tip and the outlet;

an actuator configured to translate the extractor with respect to the capture substrate, the actuator configured to place the extractor tip for fluid communication with a target well of the set of wells; and

an extraction subsystem configured to;

receive the image dataset;

control the actuator to position the extractor tip for fluid communication with the target well, based on the image dataset; and

in response to achieving positioning of the extractor tip for fluid communication with the target well, control the retriever subsystem to extract a target material from the target well.

2. The system of claim 1, wherein a cross section of each of the set of wells defines a polygon.

3. The system of claim 1, wherein the capture substrate further comprises a manifold defining a manifold inlet and a manifold outlet, wherein the manifold inlet and the manifold outlet are fluidly coupled to each well in the set of wells.

4. The system of claim 1, wherein the imaging system comprises:

an optical sensor;

a light source configured to transmit light toward the target well;

a lens defining an optical axis substantially normal the broad face, wherein the lens is configured to perform at least one of focusing light onto the target material and directing light from the target material to the optical sensor; and

a focus actuator configured to translate the lens substantially along the optical axis.

5. The system of claim 1, wherein the extractor comprises a capillary tube configured for application of a capillary force to the target material.

6. The system of claim 1, wherein the extractor comprises a pump coupled to the outlet and configured to apply a removal force in coordination with operation of the extraction subsystem.

7. The system of claim 6, wherein the extraction subsystem comprises instructions for repeated aspiration and delivery of content from and to the target well.

8. The system of claim 1, wherein the void of the extractor terminates at an aperture of the extractor tip, the aperture defining a width between 10 microns and 50 microns.

9. The system of claim 1, wherein the target material comprises at least one of a cell, a functionalized particle, and content derived from a biological sample.

10. The system of claim 1, further comprising a capture receptacle, wherein the extractor tip is operable to move the target material from the target well to the capture receptacle.

11. A system for cell retrieval from a capture substrate comprising a set of wells distributed across a broad face of the cell capture substrate, the system comprising:

a retriever subsystem comprising an extractor comprising an extractor tip, an outlet, and a void defined between the extractor tip and the outlet;

a positioning subsystem configured to position the extractor with respect to the capture substrate, the positioning subsystem configured to place the extractor tip for communication with a target well of the set of wells; and

an extraction subsystem configured to;

guide the positioning subsystem to position the extractor tip for communication with at least one target well; and

in response to achieving positioning of the extractor tip for fluid communication with the target well, control the retriever subsystem to extract a target material from the at least one target well.

12. The system of claim 11, wherein the target material comprises at least one of a cell, a functionalized particle, and content derived from a biological sample.

13. The system of claim 12, wherein a cross section of each of the set of wells defines a hexagon.

14. The system of claim 13, wherein the extraction subsystem is configured for repeated aspiration and delivery of content from and to the at least one target well.

15. The system of claim 11, wherein the extractor comprises a capillary tube configured for application of a capillary force to the target material.

16. The system of claim 11, wherein the extractor comprises a pump coupled to the outlet and configured to apply a removal force in coordination with operation of the extraction subsystem.

17. The system of claim 11, further comprising a substrate actuator comprising a platform, the substrate actuator configured to translate the capture substrate relative to the retriever subsystem.

18. The system of claim 11, further comprising an imaging subsystem configured to sample an image data dataset of one or more of the set of wells, wherein the extraction subsystem is further configured to receive the image dataset and based on the image dataset, control the positioning subsystem to position the extractor tip for fluid communication with the target well.

19. The system of claim 18, wherein the imaging subsystem comprises:

an optical sensor;

a light source configured to transmit light toward the target well;

a lens defining an optical axis substantially normal the broad face, wherein the lens is configured to perform at least one of focusing light onto the target material and directing light from the target material to the optical sensor; and

a focus actuator configured to translate the lens substantially along the optical axis.

20. The system of claim 11, wherein the set of wells is divided into a set of subsets of wells, wherein each subset of wells is circumscribed by a closed curve, and an inner diameter of the extractor tip is smaller than a largest chord of the closed curve.