System for capturing and analyzing cells

• US 9,707,562 B2
• Filed: 03/18/2016
• Issued: 07/18/2017
• Est. Priority Date: 03/13/2013
• Status: Active Grant

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Denovo Sciences, Inc. (Bio-Rad Laboratories Incorporated)
• Inventors: Sepehri, Saedeh Javdani, Handique, Kalyan, Gleason, Kyle, Payne, Austin, Gogoi, Priyadarshini, Siemer, Christopher, Zhou, Yi
• Primary Examiner(s): KWAK, DEAN P
• Application Number:
  US15/074,054
• Publication Number:
  US 20160199838A1
• Time in Patent Office: 487 Days
• Field of Search: 422 73, 422400, 422401, 422502-504, 422554
• 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 system for processing a biological sample, comprising:

a reservoir, including a reservoir inlet and a reservoir outlet, configured to receive at least one of the biological sample and a process fluid;

a fluid delivery module including a cylindrical cartridge having a set of chambers that hold the at least one fluid for processing the biological sample and an actuator coupled to a piercer configured to access at least one chamber of the set of chambers, wherein the cylindrical cartridge is rotatable about an axis to align a chamber of the set of chambers with the piercer, to facilitate flow of contents of the chamber into the reservoir inlet, and wherein the set of chambers is distributed about and offset from the axis;

a manifold configured to receive and deliver at least one of the biological sample and the process fluid from the reservoir to a biological sample substrate, the manifold comprising;

a broad surface comprising a central region configured to receive the biological sample substrate during operation of the system; and

a set of openings defined at the central region, the set of openings comprising;

a first subset of openings configured to transmit fluid flow to the biological sample substrate; and

a second subset of openings configured to receive the fluid flow from the biological sample substrate, wherein the first and the second subsets of openings are symmetrically opposed across a manifold axis;

a manifold inlet configured at an upstream end of the broad surface, the manifold inlet coupled to the reservoir and to the first subset of openings, and the manifold inlet configured to transmit flow from the reservoir to 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; and

a magnet proximal the reservoir, wherein in a first configuration, the magnet retains a concentration of contaminating particles from the biological sample when a flow of the biological sample is driven from the reservoir in a forward direction to the manifold inlet.

2. The system of claim 1, wherein the manifold axis is one of:

a longitudinal manifold axis and a lateral manifold axis.

3. The system of claim 2, wherein a first and a second vector through the first and the second subset of openings, respectively, are each substantially parallel with the manifold axis.

4. The system of claim 3, wherein the central region is a recessed region configured to provide a flow path from the first subset of openings to the second subset of openings, across the biological sample substrate.

5. The system of claim 4, further comprising an adhesive layer configured to couple the biological sample substrate to the manifold, within the recessed region of the manifold, while maintaining a gap between the biological sample substrate and the manifold, wherein the gap is configured to receive the fluid flow along the flow path.

6. The system of claim 1, wherein the manifold further comprises a first fluidic pathway configured for a first downstream flow from the manifold inlet to the first subset of openings, the first fluidic pathway branching to a first set of branches along the first downstream flow, and the first set of branches branching to a second set of branches along the first downstream flow, wherein each of the second set of branches is fluidically coupled to an opening of the first subset of openings.

7. The system of claim 6, wherein the manifold further comprises a second fluidic pathway configured for a second downstream flow from the second subset of openings to the manifold outlet, the second fluidic pathway comprising a third set of branches converging to a fourth set of branches along the downstream flow, and the fourth set of branches converging to a single channel along the downstream flow to the manifold outlet, wherein the each of the third set of branches is fluidically coupled to an opening of the second subset of openings.

8. The system of claim 1, wherein a fluidic network of the manifold is configured to re-direct a downstream flow from a first direction to a second direction to a third direction, the first and the third directions substantially parallel the longitudinal manifold axis, and the second direction across the biological sample substrate and substantially perpendicular the longitudinal manifold axis.

9. The system of claim 1, wherein the first and the second subsets of openings each comprise more than one opening, and wherein the first and the second subsets of openings are different.

10. The system of claim 1, wherein the manifold outlet is coupled to a waste chamber, the waste chamber coupled to a pump configured to provide positive pressure to enable fluid flow in a reverse direction through the manifold, and negative pressure to enable fluid flow in a forward direction through the manifold.