System for generating droplets with pressure monitoring
System, including methods, apparatus, and kits, for forming emulsions. In an exemplary method of generating droplets, a device may be selected that includes a plurality of emulsion-formation units each including a sample well, a continuous-phase well, a droplet well, and a channel network that fluidically interconnects the wells and creates a droplet-generation region. A discrete volume of sample-containing fluid may be placed into the sample well of each emulsion-formation unit, and a discrete volume of continuous-phase fluid into the continuous-phase well of each emulsion-formation unit. Pressure may be applied to the device with a fluidics assembly after the step of placing, such that the plurality of emulsion-formation units generate droplets in parallel with one another. A pressure signal may be detected from the fluidics assembly. Application of the pressure may be stopped when the pressure signal indicates that a sample well is empty.
1. A method of generating droplets, the method comprising:
selecting a device including a plurality of emulsion-formation units each including a sample well, a continuous-phase well, a droplet well, and a channel network that fluidically interconnects the wells and creates a droplet-generation region;
placing a discrete volume of sample-containing fluid into the sample well of each emulsion-formation unit, and a discrete volume of continuous-phase fluid into the continuous-phase well of each emulsion-formation unit;
applying pressure to the device with a fluidics assembly after the step of placing, such that in each emulsion-formation unit (a) sample-containing fluid flows from the sample well to the droplet-generation region, (b) continuous-phase fluid flows from the continuous-phase well to the droplet-generation region, and (c) sample-containing droplets and continuous-phase fluid flow from the droplet-generation region to the droplet well, wherein the plurality of emulsion-formation units generate droplets in parallel with one another;
detecting a pressure signal from the fluidics assembly; and
stopping application of the pressure when the pressure signal indicates that a sample well is empty.
2. The method of claim 1 , wherein the device has a row of sample wells, a row of continuous-phase wells, and a row of droplet wells, and wherein the step of applying pressure includes a step of applying pressure in parallel to each well of at least one of the rows of wells, and wherein the step of stopping application of the pressure includes a step of stopping application of pressure in parallel to each well of the at least one row of wells.
3. The method of claim 1 , wherein the step of stopping application of the pressure includes a step of adjusting a valve to vent a portion of the fluidics assembly that is applying the pressure.
4. The method of claim 1 , wherein the step of applying pressure includes a step of applying negative pressure in parallel to each of the droplet wells, and wherein the step of stopping application of the pressure includes a step stopping application of the negative pressure to each of the droplet wells.
5. The method of claim 4 , wherein the fluidics assembly includes a manifold, wherein the negative pressure is applied via the manifold, and wherein the manifold is sealed to each of the droplet wells via a same gasket.
6. The method of claim 1 , wherein the fluidics assembly includes a pressure reservoir that is operatively connected to a pump, wherein the pump is configured to create negative or positive pressure in the pressure reservoir, and wherein the step of applying pressure includes a step of applying negative or positive pressure via the pressure reservoir while the pressure reservoir remains isolated from the pump.
7. The method of claim 1 , the pressure being first pressure, further comprising a step of applying second pressure after the step of stopping application, such that continuous-phase fluid is removed selectively, relative to droplets, from an emulsion present in each droplet well.
8. The method of claim 7 , wherein the second pressure includes positive pressure applied to each of the droplet wells.
9. The method of claim 7 , wherein the second pressure causes continuous-phase fluid to flow from the droplet well of each emulsion-formation unit via a channel that connects the droplet well to the corresponding droplet-generation region.
10. The method of claim 7 , wherein the step of applying first pressure is performed for a first length of time, and wherein the step of applying second pressure is performed for a second length of time that is based on the first length of time.
11. A method of generating droplets, the method comprising:
selecting a device including a plurality of separate emulsion-formation units each including a sample well, a continuous-phase well, a droplet well, and a channel network that fluidically interconnects the wells and creates a droplet-generation region;
placing a discrete volume of sample-containing fluid into the sample well of each emulsion-formation unit, and a discrete volume of continuous-phase fluid into the continuous-phase well of each emulsion-formation unit;
applying negative pressure to the droplet well of each of the emulsion-formation units via a manifold after the step of placing, with the manifold sealed to each droplet well via a same gasket, such that in each emulsion-formation unit (a) sample-containing fluid flows from the sample well to the droplet-generation region, (b) continuous-phase fluid flows from the continuous-phase well to the droplet-generation region, and (c) sample-containing droplets and continuous-phase fluid flow from the droplet-generation region to the droplet well, wherein the plurality of emulsion-formation units generate droplets in parallel with one another;
detecting a pressure signal corresponding to the negative pressure; and
stopping application of negative pressure to all of the droplet wells in parallel when the pressure signal indicates that air has entered the channel network of an emulsion-formation unit.
12. The method of claim 11 , wherein the step of stopping application includes a step of adjusting a valve to vent each droplet well.
13. The method of claim 11 , wherein the manifold is connected to a pressure reservoir that is operatively connected to a pump, wherein the pump is configured to create negative pressure in the pressure reservoir, and wherein the step of applying negative pressure is performed via the pressure reservoir while the pressure reservoir remains isolated from the pump.
14. The method of claim 11 , further comprising a step of applying positive pressure in parallel to each of the droplet wells after the step of stopping application, such that continuous-phase fluid is removed selectively, relative to droplets, from an emulsion present in each droplet well.
15. The method of claim 14 , wherein the positive pressure causes continuous-phase fluid to flow from the droplet well of each emulsion-formation unit via a channel that connects the droplet well to the corresponding droplet-generation region.
16. The method of claim 14 , wherein the step of applying negative pressure is performed for a first length of time, and wherein the step of applying positive pressure is performed for a second length of time that is based on the first length of time.