Automated detection and repositioning of micro-objects in microfluidic devices

Patent metadata

Field	Value
Publication number	US-10832404-B2
Application number	US-201816000385-A
Country	US
Kind code	B2
Filing date	Jun 5, 2018
Priority date	Dec 9, 2014
Publication date	Nov 10, 2020
Grant date	Nov 10, 2020

Abstract

Methods are provided for the automated detection of micro-objects in a microfluidic device. In addition, methods are provided for repositioning micro-objects in a microfluidic device. In addition, methods are provided for separating micro-objects in a spatial region of the microfluidic device.

First claim

What is claimed:

1. A method of re-positioning a micro-object in a microfluidic device comprising a plurality of sequestration pens, the method comprising:
identifying a first micro-object of a set of micro-objects disposed within the microfluidic device;
computing one or more trajectories for the first micro-object, each trajectory comprising a path connecting the first micro-object, at a first location with a second location within the microfluidic device, wherein the first or second location is located in a sequestration pen of the plurality of sequestration pens disposed within the microfluidic device;
selecting a trajectory from the one or more trajectories computed for the first micro-object, wherein the length of the selected trajectory is minimized; and
re-positioning the micro-object by moving the micro-object along its selected trajectory.

2. The method of claim 1 , further comprising:
identifying a second micro-object of the set of micro-objects disposed within the microfluidic device;
computing one or more trajectories for the second micro-object, each trajectory comprising a path connecting the second micro-object, at a first location with a second location within the microfluidic device, wherein the first or second location is located in a sequestration pen of the plurality of sequestration pens disposed within the microfluidic device;
selecting a trajectory from the one or more trajectories computed for the second micro-object; and
re-positioning the second micro-object by moving the second micro-object along the selected trajectory from the one or more trajectories computed for the second micro-object.

3. The method of claim 2 , wherein the movement of the first micro-object and the second micro-object is synchronized.

4. The method of claim 2 , further comprising identifying the micro-object using machine learning techniques.

5. The method of claim 1 , further comprising:
computing a density value associated with the set of micro-objects; and
computing the one or more trajectories based, at least in part, on the density value associated with the set of micro-objects.

6. The method of claim 5 , further comprising:
determining if the density value exceeds a threshold value; and
computing, for at least one micro-object of the set of micro-objects, one or more trajectories connecting the micro-object with one or more sequestration pens of the set of sequestration pens.

7. The method of claim 5 , further comprising:
determining that the density value does not exceed a threshold value; and
computing, for at least one sequestration pen of the set of sequestration pens, one or more trajectories connecting the sequestration pen with one or more micro-objects of the set of micro-objects.

8. The method of claim 1 , wherein the second location is located in a sequestration pen of the plurality of sequestration pens disposed within the microfluidic device.

9. The method of claim 8 , further comprising identifying a set of sequestration pens, wherein said identifying comprises identifying empty sequestration pens amongst the plurality of sequestration pens.

10. The method of claim 8 , wherein the sequestration pen comprises an isolation region and a connection region which connects the isolation region to a microfluidic channel of the microfluidic device.

11. The method of claim 10 , wherein the selected trajectory moves from the microfluidic channel, through the connection region, and ends in the isolation region of the sequestration pen.

12. The method of claim 1 , further comprising: calculating a sum of lengths of one or more trajectories computed for the first micro-object; and minimizing the sum of the lengths of the one or more trajectories wherein minimizing the sum of the lengths comprises using at least one of the following: a greedy algorithm, a heuristics-based algorithm, a non-linear algorithm, and a constrained search.

13. The method of claim 1 , wherein selecting a trajectory of the one or more trajectories computed for the first micro-object, further comprises determining whether a trajectory of the one or more computed trajectories exceeds a pre-determined length.

14. The method of claim 1 , wherein re-positioning the micro-object comprises accelerating the micro-object from an initial velocity to a traveling velocity over a first time period.

15. The method of claim 14 , wherein re-positioning the micro-object comprises decelerating the micro-object from the traveling velocity to a final velocity over a second time period.

16. The method of claim 1 , wherein the first or second location is within a microfluidic channel of the microfluidic device.

17. The method of claim 16 , wherein the first location is within a sequestration pen of the plurality of sequestration pens and the second location is within the microfluidic channel of the microfluidic device.

18. The method of claim 1 , wherein the first location is within a first sequestration pen of the microfluidic device and the second location is within a second sequestration pen of the microfluidic device.

19. The method of claim 1 , wherein the microfluidic device comprises a dielectrophoresis (DEP) configuration.

20. The method of claim 19 , wherein re-positioning the micro-object comprises activating patterns of DEP electrodes within the microfluidic device, thereby generating DEP forces that move the at least one micro-object.

21. A method of re-positioning a first micro-object in a microfluidic device comprising a plurality of sequestration pens, the method comprising:
identifying a first micro-object disposed within the microfluidic device;
computing one or more trajectories for the first micro-object, each trajectory of the one or more trajectories comprising a path connecting the first micro-object at a first location with a second location within the microfluidic device, wherein the first or second location is located in a sequestration pen of the plurality of sequestration pens;
selecting a trajectory from the one or more trajectories for the first micro-object by determining whether the trajectory exceeds a pre-determined maximum length; and
re-positioning the first micro-object by moving the first micro-object along its selected trajectory.

22. The method of claim 21 , further comprising:
identifying a second micro-object disposed within the microfluidic device;
computing one or more trajectories for the second micro-object, each trajectory comprising a path connecting the second micro-object at a first location with a second location within the microfluidic device, wherein the first or second location is located in a sequestration pen of the plurality of sequestration pens disposed within the microfluidic device;
selecting a trajectory from the one or more trajectories computed for the second micro-object; and
re-positioning the second micro-object by moving the second micro-object along its selected trajectory.

23. The method of claim 22 , wherein the first micro-object and the second micro-object are moved such that the movement is synchronized.

24. The method of claim 21 , further comprising:
computing a density value associated with a set of micro-objects comprising the first micro-object and the second micro-object; and
computing the one or more trajectories based, at least in part, on the density value associated with the set of micro-objects.

25. The method of claim 21 , further comprising selec

Assignees

• Berkeley Lights Inc

Inventors

• Du Fenglei
• Lundquist Paul M
• Tenney John A
• Lionberger Troy A

Classifications

• B01L2400/0427

  Electrowetting · CPC title

• B01L2300/0645

  Electrodes · CPC title

• B01L3/50273 Primary

  characterised by the means or forces applied to move the fluids · CPC title

• G01N2015/0038

  Investigating nanoparticles · CPC title

• G06T2207/30024

  Cell structures in vitro; Tissue sections in vitro · CPC title