Compositions and methods for passive optical barcoding for multiplexed assays
Inventors
Kim, Jeffrey • Nguyen, Anh Tuan • Miller, Brandon
Assignees
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Publication Number
US-12276659-B2
Publication Date
2025-04-15
Expiration Date
Abstract
Compositions comprising multiple hydrogel particles having substantially the same diameter, but with each subgrouping of particles from the multiple hydrogel particles having different associated values for one or more passive optical properties that can be deconvoluted using cytometric instrumentation. Each hydrogel particle from the multiple hydrogel particles can be functionalized with a different biochemical or chemical target from a set of targets. A method of preparing hydrogel particles includes forming droplets and polymerizing the droplets, with optional functionalization.
Core Innovation
The disclosure describes hydrogel particle compositions for demultiplexing one or more targets based on an encoded optical property. The composition includes a plurality of populations of hydrogel particles, where each population has substantially the same average particle diameter, and each population is encoded with a different forward and/or side scatter optical property so that the populations are optically distinguishable from one another by cytometric instrumentation.
The described populations further include different functionalized ligands configured to form particle target combinations with targets selected from chemical, cellular, or biochemical targets. The hydrogel particle populations are configured to enable simultaneous demultiplexing of the one or more targets, and optional fluorescence is described as an additional demultiplexing property in some embodiments.
The disclosure also describes encoding strategies in which passive optical properties are tuned while maintaining identically sized particle populations. The optical properties can be deconvoluted using forward scatter and/or side scatter signals associated with the different hydrogel particle subpopulations, including an example where side scatter changes due to encapsulated nanoparticles while forward scatter remains generally unchanged.
Claims Coverage
The partial content provides one independent claim directed to a composition for demultiplexing based on encoded optical properties, using multiple hydrogel particle populations that share substantially the same particle diameter but differ in forward and/or side scatter properties and functionalized ligands. Dependent claims within the provided set further specify demultiplexing modalities and quantitative constraints, including optional fluorescence and constraints relating to refractive index and particle diameter.
Demultiplexing composition with identically sized hydrogel populations encoded by forward and/or side scatter
A composition comprising a plurality of populations of hydrogel particles, each population having substantially the same average particle diameter, wherein each population comprises a different forward and/or side scatter optical property so as to be optically distinguishable from other populations within the composition.
Functionalized ligand-defined particle target combinations
Each population of hydrogel particles comprises a different functionalized ligand configured to form a particle target combination with a target selected from a group consisting of a chemical, cellular, or biochemical target.
Simultaneous demultiplexing of one or more targets
Each population of the plurality of hydrogel particles is configured to allow for simultaneous demultiplexing of the one or more targets.
The independent claim coverage centers on simultaneous demultiplexing using multiple hydrogel particle populations that are identically sized but optically distinguishable by different forward and/or side scatter properties, with each population carrying a different functionalized ligand to form particle-target combinations for chemical, cellular, or biochemical targets.
Stated Advantages
Enables simultaneous demultiplexing of one or more targets using optically distinguishable hydrogel particle populations.
Documented Applications
Demultiplexing one or more targets selected from chemical, cellular, or biochemical targets in multiplexed assays using cytometric instrumentation with encoded forward and/or side scatter optical properties.
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