Chromatin immunocapture devices and methods of use

Patent metadata

Field	Value
Publication number	US-11566275-B2
Application number	US-202016827512-A
Country	US
Kind code	B2
Filing date	Mar 23, 2020
Priority date	Mar 3, 2014
Publication date	Jan 31, 2023
Grant date	Jan 31, 2023

Abstract

This application provides fluidic devices, such as microfluidic devices, which can be used for the creation and/or manipulation of droplets in droplet-based microfluidic systems, as well as systems and methods for using the same. The microfluidic devices can be used to generate droplets, extract or inject volume to droplets, and/or split droplets. Also provided are methods for generating nucleosomes, and isolated DNA from nucleosomes (or from non-nucleosomes), for example using the disclosed devices.

First claim

We claim:

1. A device comprising:
a main microchannel defining a main fluid flow path and having a first opening;
a first microchannel defining a first fluid flow path, the first fluid flow path being in fluidic communication with the main fluid flow path via the first opening, the first microchannel forming a first angle relative to the main microchannel, the first angle being less than 90 degrees;
a second microchannel defining a second fluid flow path, the second fluid flow path being in fluidic communication with the main fluid flow path via the first opening and in fluidic communication with the first fluid flow path, the second microchannel forming a second angle relative to the main microchannel, the second angle being less than 90 degrees; and
an electric field generator positioned adjacent the main fluid flow path at the location of the first opening,
wherein the first and second microchannel form a third angle relative to one another, the third angle being between 60 and 135 degrees, and
wherein the electric field generator comprises at least one source channel and at least one ground channel, wherein:
the at least one source channel comprises an electrode at an end of the source channel; and
the at least one ground channel comprises an electrode at an end of the ground channel.

2. The device of claim 1 , further comprising:
main fluid control member configured to control the flow of a first fluid in the main fluid flow path; and
at least one additional fluid control member configured to control the flow of a second fluid in the first and second fluid flow paths.

3. The device of claim 1 , wherein the electric field generator is connected to a power supply.

4. The device of claim 3 , wherein height of the source channel is 40 μm, and/or the width of the source channel is 100 μm.

5. The device of claim 3 , wherein the power supply is an AC power supply.

6. The device of claim 1 , wherein the electrode at the end of the source channel is adjacent to the main fluid flow path at the location of the first opening.

7. The device of claim 1 , wherein the electric field generator is configured to produce an electric field at the location of the first opening when an electrolytic solution is connected to the source channel and the ground channel.

8. The device of claim 7 , wherein the electrolyte solution comprises NaCl.

9. The device of claim 8 , wherein the NaCl concentration is 0.5 M.

10. The device of claim 1 , further comprising a droplet-formation channel defining a droplet-forming fluid flow path, the droplet-forming fluid flow path being in fluidic communication with the main fluid flow path via a second opening, the droplet-formation channel forming a T-junction with the main microchannel.

11. The device of claim 1 , wherein:
(a) the height of the main microchannel, first microchannel, second microchannel, droplet-formation channel, and/or opening is 10 μm to 200 μm; and/or
(b) the width of the main microchannel, first microchannel, second microchannel, droplet-formation microchannel, and/or opening is 10 μm to 200 μm.

12. The device of claim 11 , wherein the height of the main microchannel, first microchannel, second microchannel, and/or droplet-formation channel is 30 μm to 50 μm.

13. The device of claim 11 , wherein:
the width of the main microchannel, first microchannel, and/or droplet-formation channel is 30 μm to 50 μm;
the width of the second microchannel is 10 to 50 μm; and/or
the width of the opening is 5 to 30 μm.

14. The device of claim 11 , wherein the height of the main microchannel, first microchannel, second microchannel, droplet-formation microchannel, and/or opening is 40 μm.

15. The device of claim 11 , wherein the width of the main microchannel, first microchannel, second microchannel, and/or droplet-formation microchannel is 40, 25, or 15 μm.

16. The device of claim 1 , wherein the third angle is 75° to 115°.

17. The device of claim 16 , wherein the third angle is 90°.

18. The device of claim 1 , wherein the first or second angle is 20° to 60°.

19. The device of claim 18 , wherein the first and/or second angle is 45°.

Assignees

• Univ Illinois
• Mayo Found Medical Education & Res

Inventors

• Bailey Ryan
• Tice Joshua
• Ordog Tamas
• Lee Jeong Heon
• Graybill Richard Martin
• Xu Yi
• Doonan Steven

Classifications

• C12Q1/6806 Primary

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title

• B01L2200/0673

  Handling of plugs of fluid surrounded by immiscible fluid · CPC title

• C12Q2565/629

  being a microfluidic device · CPC title

• C12Q1/6844

  Nucleic acid amplification reactions · CPC title

• B01L3/502784

  specially adapted for droplet or plug flow, e.g. digital microfluidics · CPC title