System for capturing and analyzing cells

• US 9,821,311 B2
• Filed: 02/14/2017
• Issued: 11/21/2017
• 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: Handique, Kalyan, Gleason, Kyle, Payne, Austin, Gogoi, Priyadarshini, Siemer, Christopher, Zhou, Yi, Sepehri, Saedeh Javdani
• Primary Examiner(s): KWAK, DEAN P
• Application Number:
  US15/432,050
• Publication Number:
  US 20170151569A1
• Time in Patent Office: 280 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;

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

wherein the reservoir is configured to receive the biological sample and to receive at least one fluid from the fluid delivery module at the reservoir inlet;

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

a broad surface comprising a central region;

a recessed region configured to receive the biological sample substrate, the recessed region defined proximal the central region; and

a set of openings defined proximal the recessed 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 recessed region provides a flow path from the first subset of openings to the second subset of openings, across the biological sample substrate;

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, a pump drives flow of the biological sample from the reservoir in a forward direction to the manifold inlet, and the magnet retains a concentration of contaminating particles from the biological sample.

2. The system of claim 1, wherein the first and the second subsets of openings are symmetrically opposed across a manifold axis of the manifold.

3. The system of claim 1, wherein the manifold further comprises:

a first fluidic pathway from the manifold inlet to the first subset of openings, the first fluidic pathway comprising a first set of branches, wherein each of the first set of branches is fluidically coupled to an opening of the first subset of openings; and

a second fluidic pathway from the second subset of openings to the manifold outlet, the second fluidic pathway comprising a second set of branches converging to a single channel, wherein each of the second set of branches is fluidically coupled to an opening of the second subset of openings.

4. 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 direction substantially parallel the longitudinal manifold axis, the second direction substantially parallel the vertical manifold axis as the downstream flow is transmitted from the first subset of openings to the biological sample substrate, and the third direction across the biological sample substrate from the first subset of openings to the second subset of openings.

5. The system of claim 1,

wherein the first subset of openings is configured to transmit the fluid flow to a first broad surface of the biological sample substrate, and the system further comprising

an adhesive layer configured to be received at the manifold and a second broad surface of the biological sample substrate, thereby coupling the biological sample substrate to the manifold, wherein the second broad surface is opposite the first broad surface of the biological sample substrate.

6. The system of claim 1,

wherein the recessed region is a multi-tier recessed region comprising a plurality of recessed sub-regions,

wherein each tier of the multi-tier recessed region is defined at a different depth of the manifold, and

wherein each recessed sub-region is associated with a different tier of the multi-tier recessed region.

7. The system of claim 6,

further comprising a fluid delivery region defined at a first recessed sub-region of the plurality of recessed sub-regions,

wherein the fluid delivery region is raised relative to the first recessed sub-region, and

wherein fluid delivery region base dimensions are smaller than first recessed sub-region base dimensions.

8. The system of claim 7, wherein the set of openings are defined at the fluid delivery region.

9. The system of claim 8, further comprising a hermetic sealer positioned proximal the first recessed sub-region and the sides of the fluid delivery region.

10. The system of claim 7, wherein the biological sample substrate is received at a second recessed sub-region of the plurality of recessed sub-regions, and wherein the second recessed sub-region is elevated relative the first recessed sub-region.