Droplet-based particle sorting
The present invention relates to droplet-based particle sorting. According to one embodiment, a droplet microactuator is provided and includes: (a) a suspension of particles; and (b) electrodes arranged for conducting droplet operations using droplets comprising particles. A method of transporting a particle is also provided, wherein the method includes providing a droplet comprising the particle and transporting the droplet on a droplet microactuator.
1. A method of providing a droplet comprising a single particle, the method comprising:
(a) providing a droplet comprising a suspension of particles on an electrowetting droplet actuator surrounded by an oil medium;
(b) dispensing from the droplet of (a) to provide a dispensed droplet, wherein the dispensing is electrowetting-mediated without the use of dielectrophoresis;
(c) determining whether the dispensed droplet on the electrowetting droplet actuator comprises a single particle; and
(d) sorting the dispensed droplet on the electrowetting droplet actuator based on the results of the determining step using electrowetting-mediated droplet operations.
2. The method of claim 1 further comprising repeating steps 23 ( b ), 23 ( c ), and 24 ( d ).
3. The method of claim 1 wherein the dispensing is electrode-mediated.
4. The method of claim 1 wherein the dispensing is dielectrophoresis-mediated.
5. The method of claim 1 wherein the oil medium is substantially immiscible with the droplet.
6. The method of claim 1 , further wherein the single particle comprises a target particle type, and wherein step 23 ( c ) further comprises detecting the target particle type of the single particle in the dispensed droplet based on a property of and/or signal detected from the dispensed droplet.
7. The method of claim 1 further comprising combining a dispensed droplet comprising multiple particles with the suspension of particles.
8. The method of claim 1 further comprising:
(a) splitting a dispensed droplet comprising multiple particles to yield two or more daughter droplets; and
(b) analyzing one or more of the daughter droplets for the presence of a single particle.
9. The method of claim 1 further comprising:
(a) combining dispensed droplet comprising multiple particles a buffer droplet;
(b) splitting a dispensed droplet comprising multiple particles to yield two or more daughter droplets; and
(c) analyzing one or more of the daughter droplets for the presence of a single particle.
10. The method of claim 1 further comprising transporting a dispensed droplet not comprising a single particle back to the suspension of particles using electrowetting-mediated droplet operations.
11. The method of claim 1 wherein a dispensed droplet lacking particles is discarded.
12. The method of claim 1 wherein single particle-containing droplets are transported to on-droplet microactuator reservoirs.
13. The method of claim 1 wherein additional droplet operations are conducted on a dispensed droplet comprising a single particle.
14. The method of claim 1 wherein a dispensed droplet comprising a single particle is further analyzed.
15. The method of claim 1 wherein step 23 ( c ) is achieved by detecting the number of particles in the droplet based on a property of and/or signal from the dispensed droplet.
16. The method of claim 15 wherein the detecting step is achieved using a sensor.
17. The method of claim 1 further comprising combining a droplet comprising a single particle with a droplet comprising a substance which may interact with the single particle.
18. The method of claim 17 wherein the substance which may interact with the single particle is selected from the group consisting of: beads, particles, microparticles, nanoparticles, drugs, proteins, peptides, antigens, toxins, and antibodies.
19. The method of claim 17 further comprising determining the presence or quantity of a substance indicative of the particle's response to the substance.
20. The method of claim 19 wherein the determining step comprises conducting an assay comprising droplet-based steps.
21. The method of claim 17 wherein the particle comprises a T cell.
22. The method of claim 1 wherein step 23 ( c ) is based on a signal detected within the dispensed droplet.
23. The method of claim 22 wherein the signal comprises a fluorescence from a molecule which binds to the particle.
24. The method of claim 22 wherein the signal comprises radioactive particles.
25. The method of claim 22 wherein the signal comprises a visual image.