Microfluidic device for storage and well-defined arrangement of droplets

• US 9,664,619 B2
• Filed: 04/28/2009
• Issued: 05/30/2017
• Est. Priority Date: 04/28/2008
• Status: Active Grant

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Rowat, Amy, Weitz, David A., Boehm, Christian, Koester, Sarah, Agresti, Jeremy
• Primary Examiner(s): Gross, Christopher M
• Application Number:
  US12/990,102
• Publication Number:
  US 20110190146A1
• Time in Patent Office: 2,954 Days
• Field of Search: None
• CPC Class Codes: B01L 2200/0621 Control of the sequence of chambers filled or emptied
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• CPC Class Codes: B01L 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics

Abstract

The present invention relates to systems and methods for the arrangement of droplets in pre-determined locations. Many applications require the collection of time-resolved data. Examples include the screening of cells based on their growth characteristics or the observation of enzymatic reactions. The present invention provides a tool and related techniques which addresses this need, and which can be used in many other situations. The invention provides, in one aspect, a tool that allows for stable storage and indexing of individual droplets. The invention can interface not only with microfluidic/microscale equipment, but with macroscopic equipment to allow for the easy injection of liquids and extraction of sample droplets, etc.

Claims

1. A method of arranging droplets in an array, comprising:

urging a first droplet through a first constriction in a storage channel into a first pot in the storage channel,

trapping the first droplet in the first pot, and

applying a pressure differential along the storage channel such that the first droplet is urged through a second constriction in the storage channel into a second pot in the storage channel that is downstream of the first pot, and a second droplet is urged through the first constriction of the storage channel into the first pot of the storage channel, wherein, upon removal of the applied pressure differential, the first droplet is trapped within the second pot of the storage channel and the second droplet is trapped within the first pot of the storage channel simultaneously.

2. The method of claim 1, further comprising:

forming a hole in the second pot; and

transferring the first droplet in the second pot into a well of a secondary device via the hole formed in the second pot.

3. The method of claim 2, wherein the secondary device comprises a microtiter plate.

4. The method of claim 1, further comprising:

determining a property of the first droplet in the second pot.

5. The method of claim 4, wherein the property comprises fluorescence.

6. The method of claim 4, wherein the property comprises a level of expression of a gene.

7. The method of claim 1, further comprising:

moving a device containing the storage channel while maintaining the first droplet within the second pot.

8. The method of claim 1, wherein the first droplet houses a chemical reaction.

9. The method of claim 1, wherein the first droplet is one of a plurality of droplets, and wherein:

the storage channel comprises a plurality of pots, and

at least about 1% of the plurality of droplets are trapped within the pots.

10. The method of claim 9, wherein the at least about 1% of the plurality of droplets are loaded into pots of the storage channel at a rate of at least about 10 droplets per second.

11. The method of claim 1, wherein the storage channel is part of a device comprising at least 100 pots.

12. The method of claim 1, wherein the first pot is in direct fluid communication with no more than two other pots in the storage channel.

13. The method of claim 1, wherein the first pot is connected to exactly two constrictions in the storage channel.

14. The method of claim 1, wherein the first droplet is a cell.

15. The method of claim 14, wherein the cell is a single-cell organism cell, a plant cell, an animal cell, a suspension cell, or an engineered cell.

16. The method of claim 1, wherein the first droplet contains a cell.

17. The method of claim 16, wherein the first droplet contains exactly one cell.

18. The method of claim 16, further comprising growing the cell in the first droplet.

19. The method of claim 16, wherein the cell is a single-cell organism cell, a plant cell, an animal cell, a suspension cell, or an engineered cell.

20. The method of claim 19, wherein the cell is a bacterium cell.

21. The method of claim 19, wherein the cell is an immune cell.

22. The method of claim 21, wherein the cell is a T cell, B cell, a macrophage, a neutrophil, a basophil, a mast cell, or an eosinophil.

23. The method of claim 19, wherein the cell is an engineered cell.

24. The method of claim 23, wherein the cell is Chinese hamster ovarian (CHO) cell or a 3T3 cell.

25. The method of claim 16, wherein the first droplet contains multiple cells of different species.

26. The method of claim 25, further comprising growing the multiple cells of different species in the first droplet.

27. The method of claim 16, further comprising carrying out a reaction of the cell in the first droplet.

28. The method of claim 16, further comprising determining a property of the cell in the second pot.

29. The method of claim 28, wherein the property comprises an amount of a chemical secreted or taken up by the cell.

30. The method of claim 16, wherein the volume of the first droplet shrinks over a period of time.

31. The method of claim 1, wherein the first droplet comprises one or more RNA, siRNA, DNA, protein, peptide, enzyme, a virus, or a combination thereof.

32. The method of claim 1, wherein the first droplet comprises a bead.

33. The method of claim 32, wherein the first droplet comprises a bead and a cell.

34. The method of claim 33, wherein the cell is a single-cell organism cell, a plant cell, an animal cell, a suspension cell, or an engineered cell.

35. The method of claim 33, further comprising growing the cell on the bead in the first droplet.

36. The method of claim 1, wherein a ratio of the diameter of the first pot to the first constriction is greater than 2:

1.

37. The method of claim 1, wherein the first pot and the second pot are adjacent and connected by the second constriction.