Compositions and methods for controllably merging emulsion droplets and sample analysis
The present disclosure in some aspects provides methods for the controlled merging of emulsion droplets, which can be used to assemble useful compositions such as droplets (e.g., stabilized micelles) containing a precise combination of analytes and/or analytical reagents. In some embodiments, disclosed herein is a method, e.g., for detecting the presence/absence, a level or amount, and/or an activity of an analyte in a sample, comprising merging two or more emulsion droplets such that an interaction between an analyte and an analyte-interacting reagent occurs in the merged droplet. The two or more emulsion droplets may be merged using a method for the controlled merging of emulsion droplets disclosed herein.
1. A method for analyzing an analyte, comprising:
contacting (i) a first emulsion comprising a first aqueous droplet in a first liquid matrix, with (ii) a second emulsion comprising a second aqueous droplet in a second liquid matrix, wherein the first aqueous droplet comprises an analyte and the second aqueous droplet comprises a reagent, under conditions that allow the first aqueous droplet to merge with the second aqueous droplet to form a merged droplet,
wherein an interaction or a reaction in the merged droplet involving the analyte and the reagent generates a detectable signal, and
wherein merging of the first and second aqueous droplets is controlled, and the controlled merging is provided by:
(i) the first aqueous droplet comprising a first redox species and the first liquid matrix comprising a first electrolyte, the second aqueous droplet comprising a second redox species and the second liquid matrix comprising a second electrolyte, wherein the first and second aqueous droplets are each contacted with an electrode to cause charge transfer between the aqueous droplet and the electrode;
(ii) the first emulsion stabilized with a first charged surfactant, and the second emulsion stabilized with a second charged surfactant of opposite charge from that of the first charged surfactant;
(iii) contacting the first emulsion with a positive electrode sufficient to cause electrostatic charging of the first aqueous droplet, and contacting the second emulsion with a negative electrode sufficient to cause electrostatic charging of the second aqueous droplet; and/or
(iv) the first aqueous droplet comprising a first magnetic particle, the second aqueous droplet comprising a second magnetic particle, wherein an external magnetic field is applied to produce an attractive force between said first magnetic particle and said second magnetic particle.
2. The method of claim 1 , wherein the detectable signal is detected for analyzing the presence or absence, an amount or concentration, and/or an activity of the analyte in a sample.
3. The method of claim 1 , wherein the first aqueous droplet has a first net ionic charge, and the second aqueous droplet has a second net ionic charge that is the opposite of the first net ionic charge.
4. The method of claim 1 , wherein the first aqueous droplet comprises a first charged surfactant and the second aqueous droplet comprises a second charged surfactant, and wherein the second surfactant is of opposite charge from the charge of the first charged surfactant.
5. The method of claim 1 , wherein the reagent comprises a fluorescent label.
6. The method of claim 1 , wherein the reagent comprises a magnetic label.
7. The method of claim 1 , wherein the contacting step comprises mixing streams of the first emulsion and the second emulsion and at least one demulsifying agent.
8. The method of claim 1 , wherein the contacting step comprises mixing streams of the first emulsion and the second emulsion and at least one surfactant.
9. The method of claim 1 , wherein the detectable signal comprises a fluorescent signal, which is optionally induced by a non-polarized excitation light, a linearly polarized excitation light, a circularly polarized excitation light, an elliptically polarized excitation light, a non-coherent excitation light, a coherent excitation light, a continuous excitation light, a pulsed excitation light, an excitation light applied at a single incident angle, or an excitation light applied at a set of incidence angles.
10. The method of claim 1 , further comprising applying an external electric field to the merged droplet to modify the distribution of the electric field vector of an evanescent wave within each droplet and thereby affect fluorescence emission.
11. The method of claim 1 , further comprising applying an external electric field to distort the merged droplet.
12. The method of claim 1 , wherein the reagent comprises a fluorescent particle and is capable of specifically binding to the analyte.
13. The method of claim 12 , wherein the first and second emulsions are mixed on a surface in an analytical chamber, thereby allowing the first and second aqueous droplets to form the merged droplet on the surface, and allowing the reagent to specifically bind to the analyte in the merged droplet.
14. The method of claim 13 , further comprising applying an excitation light at an angle sufficient to produce total internal reflection on the surface during said mixing process sufficient to cause fluorescence of the fluorescent particle, wherein said fluorescence is dependent on fluorescent particle position within the electric field vector of an evanescent wave that arises from a refractive index difference between said flat surface and the aqueous droplet.
15. The method of claim 14 , further comprising:
detecting and/or measuring the magnitude of the fluorescence emission during said mixing process; and
determining the identity of reagent used within each droplet from the pattern of fluorescent emission wavelengths from each droplet,
wherein a change in the stochastic behavior of the magnitude of the fluorescence emission provides an indication of analyte presence/absence, amount or concentration, and/or binding interaction.
16. The method of claim 12 , wherein the first and second emulsions are mixed to form a combined emulsion wherein the merged droplet is in suspension in the combined emulsion.
17. The method of claim 16 , further comprising applying an excitation light during said mixing process sufficient to cause fluorescence of the fluorescent particle, wherein said fluorescence is dependent on fluorescent particle position within the electric field vector of an evanescent wave that arises from a refractive index difference between said liquid matrix and the aqueous droplet.
18. The method of claim 17 , further comprising:
detecting and/or measuring the magnitude of the fluorescence emission during said mixing process; and
determining the identity of reagent used within each droplet from the pattern of fluorescent emission wavelengths from each droplet,
wherein a change in the stochastic behavior of the magnitude of the fluorescence emission provides an indication of analyte presence/absence, amount or concentration, and/or binding interaction.
19. A method for controlled merging of emulsion droplets, comprising contacting (i) a first emulsion comprising a first aqueous droplet in a first liquid matrix, with (ii) a second emulsion comprising a second aqueous droplet in a second liquid matrix, under conditions that allow the first aqueous droplet to merge with the second aqueous droplet to form a merged droplet, wherein the merging is controlled and is provided by:
(i) the first aqueous droplet comprising a first redox species and the first liquid matrix comprising a first electrolyte, the second aqueous droplet comprising a second redox species and the second liquid matrix comprising a second electrolyte, wherein the first and second aqueous droplets are each contacted with an electrode to cause charge transfer between the aqueous droplet and the electrode;
(ii) the first emulsion stabilized with a first charged surfactant, and the second emulsion stabilized with a second charged surfactant of opposite charge from that of the first charged surfactant;
(iii) contacting the first emulsion with a positive electrode sufficient to cause electrostatic charging of the first aqueous droplet, and contacting the second emulsion with a negative electrode sufficient to cause electrostatic charging of the second aqueous droplet; and/or
(iv) the first aqueous droplet comprising a first magnetic particle, the second aqueous droplet comprising a second magnetic particle, wherein an external magnetic field is applied to produce an attractive force between said first magnetic particle and said second magnetic particle.