Systems and methods for generation of emulsions with suitable clarity with applications of use
The inventions cover systems and methods for generation of emulsions having suitable clarity without requiring refractive index matching between emulsion components. Systems can include: a substrate including a set of openings; a reservoir facing the substrate at a first side and containing a sample fluid configured for droplet formation upon interacting with the set of openings of the substrate; and a collecting container facing the substrate at a second side and containing a set of fluid layers configured with a density gradient and suitable immiscibility characteristics. One or more components of the system(s) can support methods for emulsion generation, in relation to enabling interactions between multiple continuous phases and a dispersed droplet phase to generate clear emulsions. Applications of the inventions(s) can include performance of droplet-based digital PCR in an improved manner (e.g., without requiring implementation of correction factors based upon Poisson statistics).
1. A method of analyzing target material in a sample, the method comprising:
distributing target material across a set of partitions within a collecting container, wherein the set of partitions comprises greater than or equal to 100,000 partitions, and
upon obtaining a cross-sectional image of the collecting container containing the set of partitions, generating a count of only partitions of the set of partitions emitting signal associated with the target material.
2. The method of claim 1 , wherein the set of partitions is characterized by less than or equal to 10% occupancy by the target material.
3. The method of claim 1 , wherein generating the count omits implementation of a correction factor for partitioning error.
4. The method of claim 1 , wherein the sample comprises a mixture of fetal and maternal material.
5. The method of claim 4 , further comprising providing results for prenatal analysis of a genetic disorder based upon the count.
6. The method of claim 5 , wherein the genetic disorder comprises an aneuploidy.
7. The method of claim 1 , wherein the target material comprises at least one of: nucleic acid molecules, peptides, proteins, cells, and viral particles.
8. The method of claim 1 , wherein the set of partitions is a plurality of droplets in an emulsion.
9. The method of claim 1 , wherein the emulsion is generated by: (i) combining the sample comprising target material with a density gradient material, thereby forming a mixture; and (ii) processing the mixture with a set of fluid layers comprising an oil and water combined with a surfactant.
10. The method of claim 1 , wherein distributing the target material across the set of partitions comprises centrifuging the target material of the sample through a substrate comprising a distribution of openings, and into a set of fluid layers within the collecting container.
11. The method of claim 1 , wherein the target material comprises a plurality of nucleic acids, and wherein the method further comprises performing: (i) amplification of nucleic acids of the plurality of nucleic acids and (ii) labeling of the nucleic acids or amplicons thereof within the set of partitions.
12. The method of claim 11 , wherein the labeling comprises providing, with the sample, fluorescent probes capable of emitting signals detectable in the cross-sectional image.
13. The method of claim 11 , wherein the target material comprises more than 10,000 nucleic acids.
14. The method of claim 11 , wherein only the partitions of the set of partitions contain the nucleic acids.
15. The method of claim 1 , wherein the sample comprises single cells comprising the target material, and wherein the method further comprises performing an analysis of the single cells, based upon the count.
16. The method of claim 1 , wherein the target material comprises cell-free DNA, and wherein the method further comprises monitoring for cancer associated with the cell-free DNA, based upon the count.
17. The method of claim 8 , wherein the emulsion is characterized with clarity above a threshold level of clarity, and wherein the threshold level of clarity is 70% optical transmittance of light.