System and method for capturing and analyzing cells

• US 9,802,193 B2
• Filed: 02/14/2017
• Issued: 10/31/2017
• 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, Javdani, Saedeh Sepehri, Siemer, Christopher, Zhou, Yi
• Primary Examiner(s): COOK, LISA V
• Application Number:
  US15/431,977
• Publication Number:
  US 20170151568A1
• Time in Patent Office: 259 Days
• Field of Search: None
• 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 selected chamber defining a selected chamber interior and an aperture;

a piercer extending through the aperture into the chamber interior;

a reservoir comprising a reservoir inlet and a reservoir outlet and defining a reservoir interior;

a selected liquid fluidly coupling the selected chamber interior and the reservoir interior through the aperture and the reservoir inlet;

a manifold coupled to the reservoir at the reservoir outlet and to a cell capture device, the manifold defining a manifold interior fluidly coupling the reservoir interior to the cell capture device; and

a pump fluidly coupled to the manifold interior.

2. The system of claim 1, further comprising a mount and a cartridge rotatably coupled to the mount about an axis, the cartridge comprising a set of chambers comprising the selected chamber, wherein:

the chambers of the set of chambers are angularly displaced about the axis;

the aperture is offset from the axis; and

each chamber of the set of chambers contains a respective liquid.

3. The system of claim 2, further comprising:

a rotary actuator, wherein the cartridge is rotatably coupled to the mount by the rotary actuator; and

a piercer actuator retaining the piercer within the aperture.

4. The system of claim 2, wherein:

the cartridge comprises a cylindrical cartridge; and

the reservoir inlet is situated inferior to the aperture.

5. The system of claim 1, wherein the biological sample is contained within the reservoir interior.

6. The system of claim 5, further comprising a magnet proximal the reservoir, wherein the biological sample comprises a contaminant particle magnetically retained within the reservoir interior by the magnet.

7. The system of claim 1, further comprising a heater, wherein the cell capture device comprises:

a first surface proximal to and thermally coupled to the heater; and

a second surface opposing the first surface, the second surface fluidly coupled to the manifold interior.

8. The system of claim 7, wherein the heater comprises:

a heating substrate;

a pair of heating elements embedded in the heating substrate at opposing edges of the heating substrate;

a coolant manifold thermally coupled to the heating substrate; and

a coolant fluid contained within the coolant manifold.

9. The system of claim 8, wherein the heating substrate comprises the coolant manifold.

10. The system of claim 1, further comprising:

a level sensor configured to generate a signal upon detection of a fluid level; and

a 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 by a sample processing system, the method comprising:

providing;

a fluid delivery module comprising a first chamber and an opener, the first chamber containing a first liquid;

a reservoir; and

a cell capture device configured to capture the target cell population;

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

upon opening the first outlet, delivering the first liquid along a fluid path from the first outlet into the reservoir;

delivering the first liquid from the reservoir into the cell capture device;

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; and

delivering the cell-capture device from the sample processing system.

12. The method of claim 11, wherein the opener comprises a piercer and opening the first outlet comprises piercing the first chamber with the piercer.

13. The method of claim 11, wherein:

the fluid delivery module further comprises a cartridge comprising a set of chambers angularly displaced about an axis of the cartridge, the set of chambers comprising the first chamber; and

the method further comprises, before delivering the first liquid along the fluid path, rotating the cartridge about the axis to align the first chamber with the fluid path.

14. The method of claim 13, further comprising, after delivering the biological sample into the cell capture device, washing the target cell population captured from the biological sample at the cell capture device, wherein washing comprises:

rotating the cartridge about the axis to substantially align a second chamber of the set with the fluid path, the second chamber containing a second liquid;

opening a second outlet of the second chamber with the opener;

upon opening the second outlet, delivering the second liquid substantially along the fluid path from the second outlet into the reservoir; and

delivering the second liquid from the reservoir into the cell capture device.

15. The method of claim 11, 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.

16. The method of claim 11, further comprising, after delivering the biological sample into the cell capture device, delivering a processed volume comprising the target cell population and a set of contaminating particles from the cell capture device into the reservoir.

17. The method of claim 16, further comprising, after delivering the processed volume into the reservoir:

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.

18. The method of claim 11, wherein the fluid delivery module further comprises a cartridge comprising a set of chambers, the set of chambers comprising the first chamber, the method further comprising:

receiving information regarding a sample preparation protocol for the biological sample; and

delivering a set of liquids contained within the set of chambers into the reservoir, based upon the sample preparation protocol.

19. The method of claim 18, wherein the sample preparation protocol comprises a fluorescence in situ hybridization protocol, and delivering the set of liquids comprises:

substantially aligning a second chamber of the set with the fluid path;

opening a second outlet of the second chamber with the opener;

delivering a fluorescent tagging reagent solution substantially along the fluid path from the second chamber into the reservoir;

incubating the biological sample with the fluorescent tagging reagent solution;

substantially aligning a third chamber of the set with the fluid path;

opening a third outlet of the third chamber with the opener; and

delivering a washing liquid substantially along the fluid path from the third chamber into the reservoir.

20. The method of claim 18, wherein the sample preparation protocol comprises a polymerase chain reaction protocol, and delivering the set of liquids comprises:

substantially aligning a second chamber of the set with the fluid path;

opening a second outlet of the second chamber with the opener;

delivering a permeabilization solution substantially along the fluid path from the second chamber into the reservoir;

substantially aligning a third chamber of the set with the fluid path;

opening a third outlet of the third chamber with the opener; and

delivering a solution comprising PCR-associated materials substantially along the fluid path from the third chamber into the reservoir.