Exporting a selected group of micro-objects from a micro-fluidic device

Patent metadata

Field	Value
Publication number	US-10800652-B2
Application number	US-201816123882-A
Country	US
Kind code	B2
Filing date	Sep 6, 2018
Priority date	Oct 22, 2013
Publication date	Oct 13, 2020
Grant date	Oct 13, 2020

Abstract

A group of micro-objects in a holding pen in a micro-fluidic device can be selected and moved to a staging area, from which the micro-objects can be exported from the micro-fluidic device. The micro-fluidic device can have a plurality of holding pens, and each holding pen can isolate micro-objects located in the holding pen from micro-objects located in the other holding pens or elsewhere in the micro-fluidic device. The selected group of micro-objects can comprise one or more biological cells, such as a clonal population of cells. Embodiments of the invention can thus select a particular group of clonal cells in a micro-fluidic device, move the clonal cells to a staging area, and export the clonal cells from the micro-fluidic device while maintaining the clonal nature of the exported group.

First claim

What is claimed:

1. A process of exporting micro-objects from a micro-fluidic apparatus comprising:
an enclosure comprising a base and a micro-fluidic structure disposed on said base, wherein said enclosure defines a flow region configured to contain a liquid medium and a plurality of holding pens,
wherein said flow region comprises a channel, wherein each holding pen of said plurality opens off of said channel and comprises an isolation region and a connection region, with said isolation region fluidically connected to said channel via said connection region, and wherein said isolation region is configured to hold a group of micro-objects and isolate said group of micro-objects from micro-objects in other holding pens of said plurality,
wherein said enclosure further comprises
an inlet and an outlet, each associated with said flow region, for respectively inputting and removing said liquid medium from said flow region, said outlet providing a passage through said enclosure; and
a first electrode, a second electrode, and an electrode activation substrate comprising DEP electrodes located at an inner surface of said flow region, said process comprising:

selecting a group of micro-objects located in one holding pen of said plurality, wherein said selected group of micro-objects comprises a plurality of biological cells from a clonal colony of biological cells;
moving said selected group of micro-objects into said channel; and
exporting said selected group of micro-objects by flowing said liquid medium in said channel, wherein said export flow of said liquid medium carries said selected group of micro-objects to said outlet and through said passage through said enclosure.

2. The process of claim 1 , wherein said plurality of biological cells is a subset of the clonal colony of biological cells.

3. The process of claim 1 , wherein said biological cells are immunological cells, cancer cells, transformed cells, or stem cells.

4. The process of claim 1 , wherein said selecting comprises determining that said group of micro-objects has a particular activity.

5. The process of claim 1 , wherein said selecting comprises determining that said group of micro-objects has a particular physical characteristic.

6. The process of claim 1 , wherein selecting said group of micro-objects and moving said selected group comprises activating said DEP electrodes in a pattern that traps said group of micro-objects.

7. The process of claim 6 , wherein said DEP electrodes are activated by directing patterned light onto said electrode activation substrate at said inner surface of said flow region.

8. The process of claim 1 , wherein said exporting comprises discarding a leading volume of said liquid medium.

9. The process of claim 1 , wherein said selected group of micro-objects is exported in a volume of said liquid medium of about 1 microliter or less.

10. The process of claim 1 , wherein said selected group of micro-objects is exported in a volume of said liquid medium of about 1 microliter to about 5 microliters.

11. The process of claim 1 further comprising, after exporting said group of micro-objects, inspecting said passage through said enclosure for any un-exported micro-objects of said selected group.

12. The process of claim 11 further comprising neutralizing any un-exported ones of said micro-objects located in said passage through said enclosure.

13. The process of claim 1 , wherein said microfluidic device further comprises an export interface configured to interface with an export device, wherein said export interface is disposed on said enclosure adjacent said passage and comprises an opening to said passage.

14. The process of claim 13 , wherein said export flow of liquid medium carries said selected group of micro-objects through said export interface and into said export device.

15. The process of claim 13 further comprising, after exporting said group of micro-objects, inspecting said passage through said enclosure, said export interface, and/or said export device for any un-exported micro-objects of said selected group.

16. The process of claim 15 further comprising neutralizing any un-exported ones of said micro-objects located in said passage through said enclosure, said export interface, and/or said export device.

17. The process of claim 1 , wherein, prior to said moving of said selected group of micro-objects into said channel, the process further comprises:
flushing said channel with an initial flow of liquid medium in said channel; and
slowing or stopping said initial flow of said liquid medium in said channel.

18. The process of claim 17 comprising stopping said initial flow of said liquid medium in said channel.

19. The process of claim 17 , wherein slowing said initial flow comprises reducing the initial flow of said liquid medium in said channel to less than 0.05 uL/sec.

20. The process of claim 17 , wherein flushing said channel comprises flowing an amount of said liquid medium through said channel at a rate of 0.05 to 5.0 uL/sec, and wherein the amount is at least 2 times the combined volume of fluid that the microfluidic apparatus and the export device are capable holding.

21. The process of claim 17 , wherein the initial flow of liquid medium in said channel is continuous.

22. The process of claim 17 , wherein the initial flow of liquid medium in said channel is pulsed, periodic, or intermittent.

23. The process of claim 1 , wherein the clonal nature of said selected group of cells is maintained during said exporting.

24. The process of claim 1 , wherein said first electrode is part of a cover of said enclosure, and wherein said second electrode and said electrode activation substrate are part of said base of said enclosure.

25. The process of claim 24 , wherein said cover is comprised by said micro-fluidic structure.

26. The process of claim 1 , wherein each holding pen of said plurality comprises a barrier disposed on an inner surface of said enclosure.

Assignees

• Berkeley Lights Inc

Inventors

• Breinlinger Keith J
• Malleo Daniele
• Mathieu Gaetan L
• Nevill J Tanner
• Slocum Alexander H
• White Mark P

Classifications

• G01N35/10

  Devices for transferring samples {or any liquids} to, in, or from, the analysis apparatus, e.g. suction devices, injection devices {(G01N35/0099 takes precedence)} · CPC title

• G01N33/54326

  Magnetic particles · CPC title

• G01N33/543 Primary

  with an insoluble carrier for immobilising immunochemicals · CPC title

• G01N33/5023

  on expression patterns · CPC title

• G01N27/44791

  Microapparatus (sample containers with integrated microfluidic structures B01L3/5027) · CPC title