Flow cytometry apparatus and methods

• US 10,094,761 B2
• Filed: 06/01/2017
• Issued: 10/09/2018
• Est. Priority Date: 07/10/2014
• Status: Active Grant

• Current Assignee: Kinetic River Corp.
• Original Assignee: Kinetic River Corp.
• Inventors: Vacca, Giacomo
• Primary Examiner(s): Punnoose, Roy M
• Application Number:
  US15/611,224
• Publication Number:
  US 20170268983A1
• Time in Patent Office: 495 Days
• Field of Search: 356338
• CPC Class Codes: G01N 15/1404 Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 15/1436 the optical arrangement forming an integrated apparatus with the sample container, e.g. a flow cell
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/1006 for cytology
• CPC Class Codes: G01N 2015/1415 Control of particle position

Abstract

A particle analyzer, comprising a source of a substantially nondiffracting light beam; a flow path configured to produce in a flowcell a ribbon-like core stream having a specific cross-sectional aspect ratio; the flowcell being configured to expose a segment of the core stream to the light beam; a detector configured to receive a signal resulting from an interaction of a particle in the core stream with the light beam; a first sorting actuator connected with the flowcell, downstream of the exposed segment of core stream; a plurality of sorting channels in fluid connection with the flow path and downstream of the first actuator; the actuator having multiple actuation states, each state configured to direct at least one part of the core stream to a corresponding channel; a second sorting actuator connected with the flowcell, opposite the first actuator, and operable in coordination with the first actuator.

Claims

1. A particle analyzer, comprising:

a source of a non-Gaussian, substantially nondiffracting light beam;

a flow path configured to produce a ribbon-like core stream in a flow cell, wherein the flow cell is configured to expose a segment of the core stream to the light beam; and

a plurality of detectors configured in a linear array, wherein individual detectors of the plurality of detectors are configured to receive a signal from a corresponding portion of the core stream, the signal resulting from an interaction of a particle in the corresponding portion of the core stream with the light beam.

2. The particle analyzer of claim 1, further comprising:

a first plurality of sorting actuators connected with the flow cell, the first plurality of sorting actuators being positioned downstream of the segment of the core stream exposed to the light beam, the first plurality of sorting actuators being in substantial alignment with a largest cross-sectional dimension of the core stream; and

a plurality of sorting channels in fluid connection with the flow path and downstream of the first plurality of sorting actuators, the first plurality of sorting actuators comprising multiple actuation states, wherein each actuation state is configured to direct at least one part of the core stream to at least one corresponding sorting channel of the plurality of sorting channels.

3. The particle analyzer of claim 2, further comprising a second plurality of sorting actuators, the second plurality of sorting actuators being connected with the flow cell and being opposite the first plurality of sorting actuators, the second plurality of sorting actuators being configured to be operated in coordination with the first plurality of sorting actuators.

4. The particle analyzer of claim 1, wherein the core stream has a cross-sectional aspect ratio of at least 4 and a largest cross-sectional dimension of at least 50 micrometers.

5. The particle analyzer of claim 4, wherein the light beam is aligned with the largest cross-sectional dimension of the core stream under an angle of no more than 45 degrees.

6. The particle analyzer of claim 4, wherein the light beam is aligned substantially along the largest cross-sectional dimension of the core stream.

7. A particle analyzer, comprising:

a source of a light beam;

a flow path configured to produce a ribbon-like core stream in a flow cell, wherein the flow cell is configured to expose a segment of the core stream to the light beam;

a plurality of detectors, wherein individual detectors of the plurality of detectors are configured to receive a signal from a corresponding portion of the core stream, the signal resulting from an interaction of a particle in the corresponding portion of the core stream with the light beam;

a first plurality of sorting actuators connected with the flow cell, the first plurality of sorting actuators being positioned downstream of the segment of the core stream exposed to the light beam, the first plurality of sorting actuators being in substantial alignment with a largest cross-sectional dimension of the core stream, wherein individual sorting actuators of the first plurality of sorting actuators are configured to have multiple actuation states; and

a plurality of sorting channels in fluid connection with the flow path and downstream of the first plurality of sorting actuators, the plurality of sorting channels being configured in a two-dimensional array, wherein at least one individual sorting actuator of the first plurality of sorting actuators is selectable to operate in at least one of the multiple actuation states to direct at least one part of the core stream to at least one corresponding sorting channel of the plurality of sorting channels.

8. The particle analyzer of claim 7, further comprising a second plurality of sorting actuators, the second plurality of sorting actuators being connected with the flow cell and being opposite the first plurality of sorting actuators, the second plurality of sorting actuators being configured to be operated in coordination with the first plurality of sorting actuators.

9. The particle analyzer of claim 7, wherein the light beam is a non-Gaussian, substantially nondiffracting light beam.

10. The particle analyzer of claim 7, wherein the core stream has a cross-sectional aspect ratio of at least 4 and a largest cross-sectional dimension of at least 50 micrometers.

11. The particle analyzer of claim 10, wherein the light beam is aligned with the largest cross-sectional dimension of the core stream under an angle of no more than 45 degrees.

12. The particle analyzer of claim 10, wherein the light beam is aligned substantially along the largest cross-sectional dimension of the core stream.

13. A method comprising:

providing a particle analyzer comprising;

a source of a non-Gaussian, substantially nondiffracting light beam,

a flow path configured to produce a ribbon-like core stream in a flow cell, wherein the flow cell is configured to expose a segment of the core stream to the light beam, and

a plurality of detectors, wherein individual detectors in the plurality of detectors are configured to receive a signal from a corresponding portion of the core stream, the signal resulting from an interaction of a particle in the corresponding portion of the core stream with the light beam;

creating the ribbon-like core stream;

exposing the ribbon-like core stream to the non-Gaussian, substantially nondiffracting light beam in the flow cell;

introducing a particle in the ribbon-like core stream;

creating a signal resulting from the interaction of the non-Gaussian, substantially nondiffracting light beam with the particle in the ribbon-like core stream;

detecting the signal from the interaction of the non-Gaussian, substantially nondiffracting light beam with the particle with a detector, wherein the detector is one of the plurality of detectors; and

using the signal to determine a location of the particle across the core stream.

14. The method of claim 13, wherein the particle analyzer further comprises:

a first plurality of sorting actuators connected with the flow cell, the first plurality of sorting actuators being positioned downstream of the segment of the core stream exposed to the light beam, the first plurality of sorting actuators being in substantial alignment with a largest cross-sectional dimension of the core stream;

a plurality of sorting channels in fluid connection with the flow path and downstream of the first plurality of sorting actuators, the first plurality of sorting actuators comprising multiple actuation states, wherein each actuation state is configured to direct at least one part of the core stream to at least one corresponding sorting channel of the plurality of sorting channels;

a sorting actuator driver connected to the first plurality of sorting actuators; and

a processing unit connected to the sorting actuator driver.

15. The method of claim 14, further comprising:

transferring the signal to the sorting actuator driver through the processing unit;

using the location of the particle across the core stream to select at least one first sorting actuator, wherein the at least one first sorting actuator is one of the first plurality of sorting actuators;

bringing the at least one first sorting actuator into a desired actuation state with the sorting actuator driver in response to the signal; and

using the desired actuation state of the at least one first sorting actuator to direct a part of the core stream with the particle to a desired sorting channel.

16. The method of claim 15, wherein the particle analyzer further comprises a second plurality of sorting actuators connected to the flow cell and opposite the first plurality of sorting actuators, the second plurality of sorting actuators being connected to the sorting actuator driver and being configured to be operated in coordination with the first plurality of sorting actuators.

17. The method of claim 16, further comprising bringing at least one second sorting actuator into a desired actuation state with the sorting actuator driver in response to the signal, wherein the at least one second sorting actuator is one of the second plurality of sorting actuators.

18. The method of claim 13, wherein the core stream has a cross-sectional aspect ratio of at least 4 and a largest cross-sectional dimension of at least 50 micrometers.

19. The method of claim 18, wherein the light beam is aligned with the largest cross-sectional dimension of the core stream under an angle of no more than 45 degrees.

20. The method of claim 18, wherein the light beam is aligned substantially along the largest cross-sectional dimension of the core stream.