System and method for droplet detection

Patent metadata

Field	Value
Publication number	US-11499183-B2
Application number	US-201816022500-A
Country	US
Kind code	B2
Filing date	Jun 28, 2018
Priority date	Jun 28, 2017
Publication date	Nov 15, 2022
Grant date	Nov 15, 2022

Abstract

Systems and methods for detection of a signal from droplets of an emulsion. An exemplary system may comprise a fluid transporter having a tube with an open end for aspirating droplets, a singulator to arrange the droplets in single file and to space the single-file droplets from one another, and a detection channel in optical communication with a detector configured to detect a signal from droplets. In some embodiments, the singulator may have a channel junction at which a stream of droplets in single file is combined with a stream of spacing fluid, and a tapered spacing channel extending downstream from the channel junction toward the detection channel. In some embodiments, the fluid transporter may suck droplet-containing fluid and spacing fluid through the detection channel from respective sources. In some embodiments, droplets may be subjected to a disaggregation routine before they are passed through the detection channel.

First claim

We claim:

1. A method of droplet detection, the method comprising:
providing a microfluidic device including a flow cell defining a plane and a channel network, the channel network including a sample inlet channel, at least one spacing-fluid inlet channel, and a spacing channel that are fluidically connected to one another at a channel junction, the channel network also including a detection channel, wherein the spacing channel has a first end fluidically connected to the channel junction and a second end fluidically connected to the detection channel;
driving a spacing fluid, and a plurality of droplets in a carrier liquid, through the channel network of the flow cell using one or more positive/negative pressure sources, wherein driving (a) aligns the plurality of droplets with one another in the sample inlet channel to generate a single-file droplet stream of the plurality of droplets in the carrier liquid entering the channel junction, (b) adds the spacing fluid to the single-file droplet stream in the carrier liquid at a location in the spacing channel where a cross-sectional area of a portion of a length of the spacing channel is decreasing towards the detecting channel, and (c) passes the single-file droplet stream through the spacing channel and the detection channel:
irradiating the plurality of droplets in the detection channel using a light source; and
detecting a signal from the irradiated plurality of droplets using a detector.

2. The method of claim 1 , wherein driving includes passing the plurality of droplets through a tapered region of the sample inlet channel to align droplets of the plurality of droplets with one another.

3. The method of claim 1 , wherein a velocity of spacing fluid entering the channel junction and a velocity of the plurality of droplets entering the channel junction are the same.

4. The method of claim 2 , wherein the tapered region of the sample inlet channel defines a central axis and has an angle of taper with respect to the central axis of less than 10 degrees.

5. The method of claim 1 , wherein driving includes applying suction to the channel network downstream of the detection channel using a negative pressure source of the one or more positive/negative pressure sources.

6. The method of claim 1 , wherein the flow cell has a planar upper side opposite a planar lower side, and wherein the channel network is located between the planar upper side and the planar lower side of the flow cell.

7. The method of claim 1 , wherein at least a portion of the spacing channel tapers toward the detection channel with a progressively decreasing angle of taper, wherein the at least a portion of the spacing channel has a length measured along a long axis of the spacing channel and has a maximum width measured perpendicular to the long axis and parallel to the plane, and wherein the length is greater than the maximum width.

8. The method of claim 7 , wherein the length of the at least a portion of the spacing channel is at least ten times a minimum width of the at least a portion of the spacing channel measured parallel to the plane.

9. The method of claim 8 , wherein the maximum width is at least three times the minimum width of the at least a portion of the spacing channel.

10. The method of claim 7 , wherein the at least a portion of the spacing channel has a constant depth measured perpendicular to the plane.

11. The method of claim 7 , wherein the at least one spacing-fluid inlet channel includes a pair of spacing-fluid inlet channels defining respective long axes that form an angle of less than 50 degrees with one another.

12. The method of claim 2 , wherein the tapered region of the sample inlet channel has a length and a minimum width each measured parallel to the plane, and wherein the length of the tapered region of the sample inlet channel is at least ten times the minimum width.

13. The method of claim 7 , wherein the length is at least three times the maximum width of the at least a portion of the spacing channel.

Assignees

• Bio Rad Laboratories Inc

Inventors

• Stumbo David P
• Carman George
• Hobbs Steve
• Makarewicz Jr Anthony J
• Simonian Dmitri
• Glade David
• Oen Joshua
• Pristinski Denis
• Dzenitis John

Classifications

• B01L2300/0816

  Cards, e.g. flat sample carriers usually with flow in two horizontal directions · CPC title

• B01L2400/0487

  fluid pressure, pneumatics · CPC title

• G01N15/1434

  Optical arrangements · CPC title

• B01L2200/0673

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

• C12Q1/6851

  Quantitative amplification · CPC title