SYSTEM AND METHOD FOR CAPTURING AND ANALYZING CELLS

• US 20140272965A1
• Filed: 03/13/2014
• Published: 09/18/2014
• 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: Zhou, Yi, Handique, Kalyan, Gleason, Kyle, Payne, Austin, Gogoi, Priyadarshini, Javdani, Saedeh Sepehri, Siemer, Christopher
• Granted Patent: US 9,174,216 B2
• Time in Patent Office: Days
• US Class Current: 435/6.11
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• 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 2300/185 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: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: C12Q 1/6841 In situ hybridisation
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 2333/46 from vertebrates
• CPC Class Codes: G01N 33/56972 White blood cells
• 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
• CPC Class Codes: G01N 35/1079 with means for piercing stoppers or septums

Abstract

A system and method for capturing and analyzing cells comprising: a fluid delivery module; a reservoir configured to receive a biological sample including a target cell population and at least one fluid from the fluid delivery module; a manifold configured to receive and distribute the biological sample and at least one fluid from the reservoir into a cell capture device; a waste chamber configured to couple to the manifold; and a pump configured to couple to the waste chamber. In embodiments of the system configured to promote further purification of captured cells, the system can further comprise a magnet that enables further separation of captured cells from undesired sample materials. The system can additionally further comprise a heater configured to heat at least one fluid and/or biological sample, and a cell capture device configured to couple to the manifold, in order to facilitate capture of the target cell population.

Claims

1. A system for capturing and analyzing a target cell population from a biological sample, comprising:

a fluid delivery module including cartridge having a set of chambers that hold 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;

a reservoir, including a reservoir inlet and a reservoir outlet, configured to receive the biological sample and to receive at least one fluid from the fluid delivery module at the reservoir inlet,

wherein the cartridge of the fluid delivery module 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;

a manifold coupled to the reservoir at the reservoir outlet and configured to receive and deliver the biological sample and at least one fluid from the reservoir into a cell capture device;

a waste chamber coupled to the manifold and configured to receive waste fluid from the manifold; and

a pump coupled to at least one of the waste chamber, the manifold, and the waste chamber and configured to drive fluid flow within the manifold.

2. The system of claim 1, wherein the cartridge is a cylindrical cartridge including a set of chambers angularly displaced about the axis, and wherein the reservoir is situated inferior to the cylindrical cartridge and offset from the axis, thereby facilitating flow from the set of chambers into the reservoir.

3. The system of claim 2, wherein the cylindrical cartridge is coupled to a first actuator configured to enable identification of a rotational position of the cylindrical cartridge, and wherein the system comprises a second actuator coupled to the piercer that enables access to contents of at least one of the set of chambers, wherein the piercer includes a puncturing tip coupled to an aperture that enables fluid flow from the cylindrical cartridge to the reservoir.

4. The system of claim 1, wherein the manifold includes:

a manifold inlet coupled to the reservoir;

a set of openings including a first subset of openings configured to transmit fluid flow into the cell capture device and a second subset of openings configured to receive fluid flow from the cell capture device;

a manifold outlet that transmits waste fluid from the manifold; and

a fluid network, coupled to the manifold inlet, the set of openings, and the manifold outlet, to enable flow through the manifold.

5. The system of claim 1, wherein the waste chamber is 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.

6. The system of claim 5, further comprising a magnet proximal the reservoir, wherein, in a first configuration, the pump drives flow of the biological sample from the reservoir in a forward direction and the magnet retains a concentration of contaminating particles from the biological sample at the reservoir, thereby facilitating separation of the target cell population from other constituents of the biological sample.

7. The system of claim 1, further comprising a heater configured to heat the cell capture device at a first surface, opposed to a second surface of the cell capture device interfacing with the manifold.

8. The system of claim 7, wherein the heater includes a pair of heating elements embedded in a heating substrate at opposing edges of the heating substrate, and a plate including a fluid path configured to distribute a coolant throughout the heating substrate.

9. The system of claim 1, further comprising a bubble removal module that drives fluid in a forward direction and in a reverse direction through the cell capture device, thereby removing bubbles trapped in the cell capture device.

10. The system of claim 1, wherein at least one of the reservoir and the waste chamber comprises a level sensor configured to generate a signal upon detection of a fluid level, and processor comprising a first module that receives the signal and a second module that generates a command to control fluid delivery into the manifold, based upon the signal.

11. A method for capturing and analyzing a target cell population from a biological sample, comprising:

at a fluid delivery module, aligning a first chamber containing a buffer solution with a reservoir configured to receive the buffer solution from the first chamber;

delivering the buffer solution from the first chamber into the reservoir, and from the reservoir into a cell capture device configured to capture the target cell population;

delivering the biological sample, including the target cell population, into the cell capture device, thereby capturing the target cell population, in single-cell format, at the cell capture device;

washing the target cell population captured from the biological sample at the cell capture device, by way of the fluid delivery module;

preparing the target cell population captured at the cell capture device for analysis; and

analyzing the target cell population captured at the cell capture device.

12. The method of claim 11, further comprising driving flow of the buffer solution in a forward direction and in a reverse direction through the cell capture device, thereby removing bubbles trapped in the cell capture device.

13. The method of claim 11, further comprising reversing fluid flow within the cell capture device after delivering the biological sample into the cell capture device, thereby releasing a processed volume comprising the target cell population and a concentration of contaminating particles into the reservoir.

14. The method of claim 13, further comprising selectively binding particles of the concentration of contaminating particles to magnetic beads, and retaining the concentration of contaminating particles at the reservoir by way of a magnet proximal the reservoir, while redelivering the target cell population into the cell capture device.

15. The method of claim 14, wherein binding particles of the concentration of contaminating particles to magnetic beads comprises binding a concentration of white blood cells to streptavidin-treated magnetic beads, and wherein redelivering the target cell population into the cell capture device comprises redelivering a CTC population into the cell capture device.

16. The method of claim 13, further including tagging the concentration of contaminating particles with a marker configured to facilitate separation of the concentration of contaminating particles from the target cell population, at the reservoir.

17. The method of claim 11, wherein aligning the first chamber containing the buffer solution with the reservoir includes rotating the first chamber into alignment with the reservoir, and wherein delivering the buffer solution from the first chamber into the reservoir includes puncturing a first seal enclosing the first chamber, thereby facilitating flow of the buffer solution into the reservoir.

18. The method of claim 17, further including rotating a second chamber of the fluid delivery module into alignment with the reservoir, and wherein preparing the target cell population captured at the cell capture device for analysis includes puncturing a second seal enclosing the second chamber to facilitate flow of a processing reagent into the reservoir and into the cell capture device, with the target cell population.

19. The method of claim 11, further including positioning the cell capture device between a manifold, configured to facilitate fluid flow throughout the cell capture device, and a heater, and heating the target cell population at the cell capture device by way of the heater.

20. The method of claim 11, further including receiving information regarding a sample preparation protocol for the biological sample, wherein preparing the target cell population for analysis comprises automatically aligning chambers of a set of chambers of the fluid delivery module with the reservoir, based upon the sample preparation protocol.