System and method for capturing and analyzing cells

• US 9,610,581 B2
• Filed: 09/23/2015
• Issued: 04/04/2017
• 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:
  US14/863,191
• Publication Number:
  US 20160008814A1
• Time in Patent Office: 559 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 method for capturing and analyzing a target cell population from a biological sample, comprising:

at a fluid delivery module of a sample processing system, rotating a first chamber, of a cylindrical cartridge having a set of chambers, into alignment with a reservoir configured to receive a buffer solution from the first chamber;

delivering the buffer solution from the first chamber into the reservoir upon puncturing an inferior portion of the first chamber, 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; and

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

delivering the cell-capture device from the sample processing system, thereby preparing the target cell population captured at the cell capture device for analysis.

2. The method of claim 1, 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.

3. The method of claim 1, 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 back into the reservoir.

4. The method of claim 3, 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 in the forward direction.

5. The method of claim 4, 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 within the reservoir, and wherein redelivering the target cell population into the cell capture device comprises redelivering a CTC population into the cell capture device.

6. The method of claim 3, 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.

7. The method of claim 1, further including rotating a second chamber of the cylindrical cartridge 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.

8. The method of claim 1, 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.

9. The method of claim 8, further including delivering a coolant into a fluid network of a plate coupled to the heater, thereby cooling the target cell population at the cell capture device.

10. The method of claim 1, further including receiving information regarding a sample preparation protocol for the biological sample, wherein preparing the target cell population for analysis comprises automatically rotating chambers of the set of chambers of the fluid delivery module into alignment with the reservoir, based upon the sample preparation protocol.