Method And System For Reference-Assisted Droplet Detection, Indexing And Sorting For Assays And Diagnostics

A system for detecting, indexing, and sorting water-in-oil droplets, comprising:

• a droplet and reference-object providing module that introduces a plurality of assay droplets and a plurality of reference objects as an interspersed mixture into a microfluidic device;
• a signal detection and processing module that collects and processes signal data representing individual reference objects and assay droplets to generate respective position index for individual assay droplets by using two or more proximal reference objects of the plurality of reference objects as a position reference for a given assay droplet of the plurality of assay droplets, wherein the signal detection and processing module generates assay kinetics respectively for individual assay droplets over a predefined period of time to select a pool of droplets from the plurality of assay droplets with a predefined kinetics property;
• and
• a droplet sorting module that retrieves the selected pool of droplets for further analysis or usage.

A method for detecting and indexing droplets, comprising:

• introducing into a microfluidic device a plurality of water-in-oil droplets that comprise a plurality of assay droplets, as assay samples, and a plurality of reference objects that contain at least one kind of a reference material, wherein the reference objects and assay droplets are interspersed in a single-stream manner or a multiple single-streams manner in the microfluidic device;
• collecting signal data representing signals from individual reference objects of the plurality of reference objects and from individual assay droplets of the plurality of assay droplets in the microfluidic device at two or more time-points over a period;
• processing the collected signal data to identify the individual reference objects and the individual assay droplets, and to generate a respective position index for each of the individual assay droplets by using two or more proximal reference objects of the plurality of reference objects as a position reference, wherein the respective position index for one of the individual assay droplets is unchanged or largely unchanged over a first predefined period of time;
• and
• establishing respective assay-signal kinetics for each of the individual assay droplets over a second predefined period of time, wherein a number of the assay droplets with a predefined kinetics property are accordingly identified from a total droplet population of the plurality of assay droplets.