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US 2022/0055035 A1Published application
Microfluidic device and methods
AssigneeThe Research Foundation for The State University of New York
InventorsEric BROUZES, Martin SAUZADE
Life of this patent
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Abstract
This disclosure provides devices and methods for the isolation of single cells or particles of interest from a solution comprising a plurality of cells or a solution composed of a homogenous population of particles. Specifically, the present disclosure is directed to microfluidic devices and methods for analyzing cells in a sample. More specifically, the present disclosure provides droplet microfluidic devices and methods for using the same to obtain (trap), encapsulate, and retrieve (isolate) single cells or particles from a sample with improved efficiency.
Claims28
-19. (canceled)
A method for encapsulating a single cell in a droplet comprising:
- providing a first solution comprising at least one cell to a microfluidic device, the microfluidic device comprising:
- a first region comprising
- a microfluidic channel, wherein said microfluidic channel comprises at least one displacement element on a first inner sidewall surface of said microfluidic channel, wherein said at least one displacement element extends into the microfluidic channel toward an second inner sidewall surface of the microfluidic channel that is opposite said first inner sidewall surface, wherein said at least one displacement element comprises between three and nine tapered displacement elements, wherein each of said tapered displacement elements are aligned in series on said first inner sidewall surface of said microfluidic channel, and
- at least one trapping chamber coupled to the microfluidic channel downstream of said at least one displacement element, wherein said at least one trapping chamber comprises a first opening in said second inner sidewall of said microfluidic channel that allows the flow of fluid into said at least one trapping chamber, wherein said at least one trapping chamber comprises at least two parallel sidewalls, wherein said at least one trapping chamber comprises a second opening opposite said first opening, and wherein said second opening is connected to a trapping channel, wherein said trapping channel connects said first microfluidic channel and said second microfluidic channel, and wherein said first microfluidic channel and said second microfluidic channel are in fluid communication;
- a second region downstream of said first region comprising a second microfluidic channel;
- and
- a third region comprising a bypass channel, wherein said bypass channel adjoins the first region and the second region, wherein said bypass channel is a U-shaped channel in fluid communication with said first region and said second region,
- wherein said first solution is injected into said first microfluidic channel through a port located upstream of said tapered displacement elements, wherein said first solution flows downstream over said tapered displacement elements,
- directing said at least one cell into said at least one trapping chamber by flowing said first solution through said first microfluidic channel at a flow rate of between 6 μl/hr and 22 μl/hr,
- trapping a single cell in said at least one trapping chamber, wherein said single cell flows through said trapping chamber to said second opening in said at least one trapping chamber, wherein said single cell plugs in said second opening;
- providing a second solution to the microfluidic device, wherein said second solution flows in the same direction as the first solution toward said at least one trapping chamber and forms first interface with said first solution in said first microfluidic channel, and a second interface with said first solution across said first opening of the at least one trapping chamber;
- directing said second solution into said bypass channel and said second microfluidic channel toward said trapping channel by flowing said second solution at a flow rate of between 6 μl/hr and 22 μl/hr;
- and
- forming a droplet of said first solution comprising said single cell in said at least one trapping chamber by flowing said interface through said second microfluidic channel past said entire trapping channel.
11 more elements
A method for encapsulating a single cell in a droplet comprising:
- providing a first solution comprising at least one cell to a microfluidic device comprising:
- a first region comprising
- a microfluidic channel, wherein said microfluidic channel comprises at least one displacement element on a first inner sidewall surface of said microfluidic channel, wherein said at least one displacement element extends into the microfluidic channel toward an second inner sidewall surface of the microfluidic channel that is opposite said first inner sidewall surface, wherein said at least one displacement element comprises between three and nine tapered displacement elements, wherein each of said tapered displacement elements are aligned in series on said first inner sidewall surface of said microfluidic channel, and
- at least one trapping chamber coupled to the microfluidic channel downstream of said at least one displacement element, wherein said at least one trapping chamber comprises a first opening in said second inner sidewall of said microfluidic channel that allows the flow of fluid into said at least one trapping chamber, wherein said at least one trapping chamber comprises at least two parallel sidewalls, wherein said at least one trapping chamber comprises a second opening opposite said first opening, and wherein said second opening is connected to a trapping channel,
- a second region downstream of said first region comprising a second microfluidic channel;
- a third region comprising a bypass channel, wherein said bypass channel adjoins the first region and the second region;
- and
- a blocking rail within said at least one trapping chamber, wherein said blocking rail permits the flow of fluid through said at least one trapping chamber,
- wherein said first solution is injected into said first microfluidic channel through a port located upstream of said tapered displacement elements, wherein said first solution flows downstream over said tapered displacement elements;
- directing said at least one cell into said at least one trapping chamber by flowing said first solution through said first microfluidic channel at a flow rate of between 6 μl/hr and 22 μl/hr;
- trapping a single cell in said at least one trapping chamber, wherein said single cell flows through said first opening of said trapping chamber and contacts said blocking rail in said trapping chamber, wherein said single cell does not contact said second opening in said trapping chamber;
- providing a second solution to the microfluidic device, wherein said second solution flows in the same direction as the first solution toward said at least one trapping chamber;
- and
- directing said second solution into said at least one trapping chamber channel and said trapping channel by flowing said second solution at a flow rate of between 6 μl/hr and 22 μl/hr;
- and
- forming a droplet of said first solution comprising said single cell in said at least one trapping chamber by flowing said second solution around said blocking rail into said trapping channel.
13 more elements
A method for encapsulating a single cell in a droplet comprising:
- providing a first solution comprising at least one cell to a microfluidic device, the microfluidic device comprising:
- a first region comprising
- a microfluidic channel, wherein said microfluidic channel comprises at least one displacement element on a first inner sidewall surface of said microfluidic channel, wherein said at least one displacement element extends into the microfluidic channel toward an second inner sidewall surface of the microfluidic channel that is opposite said first inner sidewall surface, wherein said at least one displacement element comprises between three and nine tapered displacement elements, wherein each of said tapered displacement elements are aligned in series on said first inner sidewall surface of said microfluidic channel, and
- at least one trapping chamber coupled to the microfluidic channel downstream of said at least one displacement element, wherein said at least one trapping chamber comprises a first opening in said second inner sidewall of said microfluidic channel that allows the flow of fluid into said at least one trapping chamber, wherein said at least one trapping chamber comprises at least two parallel sidewalls, wherein said at least one trapping chamber comprises a second opening opposite said first opening, and wherein said second opening is connected to a trapping channel, wherein said trapping channel connects said first microfluidic channel and said second microfluidic channel, and wherein said first microfluidic channel and said second microfluidic channel are in fluid communication;
- a second region downstream of said first region comprising a second microfluidic channel;
- and
- a third region comprising a bypass channel, wherein said bypass channel adjoins the first region and the second region, wherein said bypass channel is a U-shaped channel in fluid communication with said first region and said second region,
- wherein said first solution is injected into said first microfluidic channel through a port located upstream of said tapered displacement elements, wherein said first solution flows downstream over said tapered displacement elements;
- directing said at least one cell into said at least one trapping chamber by flowing said first solution through said first microfluidic channel at a flow rate of between 6 μl/hr and 22 μl/hr;
- trapping a single cell in said at least one trapping chamber, wherein said single cell flows through said at least one trapping chamber to said second opening in said at least one trapping chamber, wherein said single cell plugs in said second opening;
- providing a second solution to the microfluidic device, wherein said second solution flows in the same direction as the first solution toward said at least one trapping chamber and forms first interface with said first solution in said first microfluidic channel;
- directing said second solution into said at least one trapping chamber by flowing said second solution at a flow rate of less than 6 μl/hr;
- and
- forming a droplet of said first solution comprising said single cell in said trapping chamber by flowing said second solution along the inner sidewalls of the at least one trapping chamber into the trapping channel, wherein the second solution surrounds the first solution containing said trapped single cell.
11 more elements
A method for encapsulating a single cell in a droplet comprising:
- providing a first solution comprising at least one cell a microfluidic device comprising:
- a first region comprising
- a microfluidic channel, wherein said microfluidic channel comprises at least one displacement element on a first inner sidewall surface of said microfluidic channel, wherein said at least one displacement element extends into the microfluidic channel toward an second inner sidewall surface of the microfluidic channel that is opposite said first inner sidewall surface, wherein said at least one displacement element comprises between three and nine tapered displacement elements, wherein each of said tapered displacement elements are aligned in series on said first inner sidewall surface of said microfluidic channel, and
- at least one trapping chamber coupled to the microfluidic channel downstream of said at least one displacement element, wherein said at least one trapping chamber comprises a first opening in said second inner sidewall of said microfluidic channel that allows the flow of fluid into said at least one trapping chamber, wherein said at least one trapping chamber is enclosed except for said first opening;
- a second region downstream of said first region comprising a second microfluidic channel;
- a third region comprising a bypass channel, wherein said bypass channel adjoins the first region and the second region;
- and
- a pressurized control channel on an outermost surface of said at least one trapping chamber,
- wherein said first solution is injected into said first microfluidic channel through a port located upstream of said tapered displacement elements, wherein said first solution flows downstream over said tapered displacement elements,
- directing said at least one cell into said at least one trapping chamber by flowing said first solution through said first microfluidic channel at a flow rate of between 6 μl/hr and 22 μl/hr,
- trapping a single cell in said at least one trapping chamber, wherein said single cell flows into said trapping chamber;
- providing a second solution to the microfluidic device, wherein said second solution flows in the same direction as the first solution toward said at least one trapping chamber and forms a first interface with said first solution in said first microfluidic channel, and a second interface with said first solution across said first opening in the at least one trapping chamber;
- directing said second solution into said bypass channel and said second microfluidic channel by flowing said second solution at a flow rate of between 6 μl/hr and 22 μl/hr;
- and
- forming a droplet of said first solution comprising said single cell in said at least one trapping chamber by flowing said first interface past said entire at least one trapping chamber.
12 more elements
Dependencies are inferred from the claim text.