System and method for capturing and analyzing cells

• US 10,350,601 B2
• Filed: 10/12/2017
• Issued: 07/16/2019
• Est. Priority Date: 03/13/2013
• Status: Active Grant

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Celsee Diagnostics, Inc. (Bio-Rad Laboratories Incorporated)
• Inventors: Handique, Kalyan, Gleason, Kyle, Payne, Austin, Gogoi, Priyadarshini, Siemer, Christopher, Zhou, Yi, Sepehri, Saedeh Javdani
• Primary Examiner(s): Kwak, Dean
• Application Number:
  US15/782,270
• Publication Number:
  US 20180036731A1
• Time in Patent Office: 642 Days
• Field of Search: 422 73, 422400, 422401, 422502-504, 422554, 436 10, 436177, 436524, 436526
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2300/06 Auxiliary integrated devices, integrated components
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0832 cylindrical, tube shaped
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/185 using a liquid as fluid
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0683 mechanically breaking a wall or membrane within a channel or chamber
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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: B01L 3/527 for a plurality of reagents
• CPC Class Codes: B01L 3/5635 connecting two containers face to face, e.g. comprising a filter
• CPC Class Codes: B01L 7/00 Heating or cooling apparatus autoclaves B01J3/04; Heat insulating devices
• CPC Class Codes: G01N 35/0098 involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C

Abstract

A system for isolating cells in at least one of single-cell format and single-cluster format, comprising a reservoir, including a reservoir inlet and a reservoir outlet, configured to receive a biological sample and to receive at least one fluid, a manifold configured to receive and deliver the biological sample and the at least one fluid from the reservoir into a biological sample substrate, the manifold comprising a broad surface comprising a central region configured to receive the biological sample substrate, a set of openings configured to enable fluid flow transmission across the biological sample substrate, a manifold inlet configured to transmit flow from the reservoir the first subset of openings, a manifold outlet configured at a downstream end of the broad surface and coupled to the second subset of openings, the manifold outlet configured to transmit waste fluid from the manifold.

Claims

1. A method comprising:

at fluid delivery module comprising a cartridge having a first chamber, a second chamber displaced from the first chamber, and a piercer, the first chamber containing a first liquid and the second chamber containing a second liquid, providing a first movement along a first non-linear path, about an axis, between the first chamber and the piercer, wherein the first chamber is offset from the axis;

opening a first outlet of the first chamber with the piercer;

upon opening the first outlet, delivering the first liquid along a fluid path from the first outlet into a cell capture device, wherein the fluid path is offset from the axis;

delivering a biological sample, including a target cell population, into the cell capture device by exerting a first set of pressure pulses on the biological sample, thereby capturing the target cell population at the cell capture device;

washing the target cell population captured from the biological sample at the cell capture device, wherein washing comprises;

providing a second movement along a second non-linear path, relative to the axis, between the second chamber and the piercer, wherein the second chamber is offset from the axis,

opening a second outlet of the second chamber with the piercer, and

delivering the second liquid along the fluid path into the cell capture device with a second set of pressure pulses; and

delivering the target cell population from the sample processing system.

2. The method of claim 1, further comprising, after delivering the biological sample into the cell capture device, delivering a processed volume comprising the target cell population from the cell capture device into a reservoir.

3. The method of claim 2, wherein:

the first set of pressure pulses are exerted in a first direction; and

delivering the processed volume into the reservoir comprises exerting a first opposing set of pressure pulses on the processed volume, the first opposing set of pressure pulses exerted in a second direction substantially opposing the first direction.

4. The method of claim 2, wherein the processed volume further comprises a set of contaminating particles, the method further comprising:

selectively binding the set of contaminating particles to a set of magnetic beads;

retaining the set of contaminating particles at the reservoir by a magnet proximal the reservoir; and

while retaining the set of contaminating particles at the reservoir, redelivering the target cell population into the cell capture device.

5. The method of claim 1, wherein:

the cell capture device is enclosed within a manifold and arranged along a manifold path defined between a manifold inlet and a manifold outlet;

the reservoir is fluidly coupled to manifold path at the manifold inlet; and

exerting the first set of pressure pulses comprises exerting negative pressure pulses at the manifold outlet.

6. The method of claim 5, further comprising, after delivering the biological sample into the cell capture device, delivering a processed volume comprising the target cell population from the cell capture device into a reservoir by exerting a set of positive pressure pulses at the manifold outlet.

7. The method of claim 1, wherein delivering the target cell population from the sample processing system comprises delivering the cell capture device, containing the target cell population, from the sample processing system.

8. The method of claim 1, wherein the target cell population is captured at the cell capture device in single-cell format.

9. The method of claim 1, further comprising driving flow of the first liquid in a forward direction and in a reverse direction through the cell capture device, thereby removing bubbles trapped in the cell capture device.