Apparatuses, systems and methods for imaging flow cytometry

Patent metadata

Field	Value
Publication number	US-10969326-B2
Application number	US-202016734676-A
Country	US
Kind code	B2
Filing date	Jan 6, 2020
Priority date	Mar 31, 2017
Publication date	Apr 6, 2021
Grant date	Apr 6, 2021

Abstract

The present disclosure provides apparatuses, systems, and methods for performing particle analysis through flow cytometry at comparatively high event rates and for gathering high resolution images of particles.

First claim

What is claimed:

1. A system, comprising:
a flow channel,
the flow channel having an inlet, and
the flow channel defining a flow path for fluid communicated therethrough;
the flow channel defining a first region located at a first distance along the flow channel from the inlet and defining a first cross-sectional profile, and a second region located at a second distance along the flow channel from the inlet and defining a second cross-sectional profile,
the first cross-sectional profile and the second cross-sectional profile differing from one another in area;
a first detection module, the first detection module being disposed so as to receive a first
signal related to illumination of the first region of the flow channel; and
a second detection module, the second detection module being disposed so as to receive a
second signal related to illumination of the second region of the flow,
wherein the first and second cross-sectional areas differ from one another and
wherein the ratio of the first cross-sectional area to the second cross-sectional area is from about 100:1 to about 1:100, excluding 1:1.

2. The system of claim 1 , wherein the first signal and the second signal differ from one another.

3. The system of claim 1 , wherein the second detection module is operable based on a signal received by the first detection module.

4. The system of claim 1 , wherein the flow channel defines a first direction of flow along the first region and a second direction of flow along the second region, the first direction of flow differing from the second direction of flow.

5. The system of claim 1 , wherein the second detection module is an image detector.

6. The system of claim 1 , wherein the first detection module is a detector.

7. The system of claim 1 , further comprising a first illumination source configured to illuminate the first region of the flow channel.

8. The system of claim 1 , wherein the first illumination source is configured to provide illumination essentially parallel to a direction of flow of the first region or perpendicular to a direction of flow of the first region.

9. The system of claim 7 , further comprising a second illumination source configured to illuminate the second region of the flow channel.

10. The system of claim 9 , wherein the second illumination source is configured to provide illumination essentially parallel to a direction of flow of the second region.

11. The system of claim 9 , wherein the second illumination source is configured to provide illumination essentially perpendicular to a direction of flow of the second region.

12. The system of claim 9 , wherein the first illumination source and the second illumination source provide different illuminations.

13. The system of claim 1 , further comprising a particle separator in fluid communication with the flow channel.

14. The system of claim 13 , wherein the particle separator is configured to divert particles based upon particle size, particle shape, particle mass, a signal associated with one or more particles, or any combination thereof.

15. The system of claim 1 , wherein the second detection module is aligned to detect a signal parallel to a direction of flow of the second region.

16. A method, comprising:
communicating a population of particles within a flow channel,
the flow channel having an inlet,
the flow channel defining a flow path for fluid communicated therethrough, and
the flow channel defining a first region located at a first distance along the flow channel from the inlet and defining a first cross-sectional profile, and defining a second region located at a second distance along the flow channel from the inlet and defining a second cross-sectional profile,
the first cross-sectional profile and the second cross-sectional profile differing from one another in area and wherein the ratio of the first cross-sectional area to the second cross-sectional area is from about 100:1 to about 1:100, excluding 1:1;
collecting, with a first detection module disposed so as to receive a first signal related to illumination of the first region of the flow channel, a first signal from a particle traversing the first region; and
based on the first signal, selectively collecting a second signal from the particle with a second detection module as the particle traverses the second region, the second signal differing from the first signal, and the second detection module being disposed so as to receive the second signal related to illumination of the second region of the flow.

17. The method of claim 16 , wherein the second detection module comprises an image detector.

18. The method of claim 16 , wherein the second detection module is aligned to detect a signal parallel to an axis of flow of the second region.

19. The method of claim 16 , wherein the first signal and the second signal differ from one another.

20. The method of claim 16 , wherein the second detection module is operable based on a signal received by the first detection module.

21. The method of claim 16 , wherein the flow channel defines a first direction of flow along the first region and a second direction of flow along the second region, the first direction of flow differing from the second direction of flow.

22. The method of claim 16 , wherein the first detection module is a detector.

23. The method of claim 16 , further comprising operating a first illumination source configured to illuminate the first region of the flow channel.

24. The method of claim 23 , wherein the first illumination source is configured to provide illumination essentially parallel to a direction of flow of the first region or perpendicular to a direction of flow of the first region.

25. The method of claim 23 , further comprising operating a second illumination source configured to illuminate the second region of the flow channel.

26. The method of claim 25 , wherein the second illumination source is configured to provide illumination essentially parallel to a direction of flow of the second region.

27. The method of claim 25 , wherein the second illumination source is configured to provide illumination essentially perpendicular to a direction of flow of the second region.

28. The method of claim 25 , wherein the first illumination source and the second illumination source provide different illuminations.

29. The method of claim 16 , further comprising operating a particle separator in fluid communication with the flow channel.

30. The method of claim 29 , wherein the particle separator is configured to divert particles based upon particle size, particle shape, particle mass, a signal associated with one or more particles, or any combination thereof.

31. A method, comprising:
flowing a population of particles through a flow channel;
with a first optical train, illuminating at least some of the population of particles at a first region of the flow channel, the first region being located at a first distance along the flow channel from the inlet and defining a first cross-sectional profile;
with a first detection module, collecting a first signal or lack thereof related to the illuminating;
with a second optical train, illuminating at least some of the population of particles at (i) a second region of the flow channel that is located at a second distance along the

Assignees

• Life Technologies Corp

Inventors

• Kaduchak Gregory
• Ward Michael

Classifications

• G01N2015/1452

  Adjustment of focus; Alignment · CPC title

• G01N2015/1445

  Three-dimensional imaging, imaging in different image planes, e.g. under different angles or at different depths, e.g. by a relative motion of sample and detector, for instance by tomography · CPC title

• G01N2015/1438

  Using two lasers in succession · CPC title

• G01N2015/1422

  Electrical focussing · CPC title

• G01N2015/142

  Acoustic or ultrasonic focussing · CPC title