Parallel microfluidic device for high throughput cell assays in microdroplets
Inventors
Kang, Wenjing • KONRY, Tania • SARKAR, Saheli
Assignees
Northeastern University Boston
Publication Number
US-12121899-B2
Publication Date
2024-10-22
Expiration Date
2039-07-24
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Abstract
A high throughput microdroplet-based system for single cell assays in microdroplets is provided. The system integrates parallel devices and switches to enable simultaneous analysis of different cells or cell combinations, or different assay conditions, on a single microfluidic chip. Interconnections between the inlets of individual devices on a common chip enable simultaneous screening of the effect of different combinations of drugs on single cells. The use of an oil inlet and microchannels with matched total flow resistance allows the synchronous generation of droplets with the same dimensions and/or volumes.
Core Innovation
The invention relates to a high throughput microdroplet-based system for single cell assays in microdroplets. This system integrates multiple parallel microfluidic devices and switches on a single chip, enabling simultaneous analysis of different cells or cell combinations, or different assay conditions, within a unified microfluidic platform. Interconnected inlets of individual devices allow for concurrent screening of various combinations of drugs on single cells. The design includes an oil inlet and microchannels that are engineered to have matched total flow resistance, facilitating the synchronous generation of droplets with uniform dimensions and volumes.
This technology addresses the limitations of prior droplet-based microfluidic platforms that lacked the ability for high throughput, simultaneous, and multiplexed screening of multiple cells, combinations, or assay conditions within a single integrated chip. Traditional systems typically required more extended analysis times and did not allow comprehensive parallel analysis of diverse cell types or responses to multiple therapeutic agents in a coordinated manner. The present system offers higher throughput by integrating larger arrays and multiplexing capabilities while maintaining droplet uniformity and ensuring scalable, parallel operation.
Additional features include arrays for microdroplet storage, allowing for real-time monitoring through time-lapse microscopy, and optional droplet sorting and merging modules to enable enrichment or further manipulation of droplets based on specific characteristics or experimental needs. This integrated approach allows for robust single cell analysis in applications such as drug screening, cell-cytokine interaction studies, and immunotherapeutic assessment at the single cell level using minimized reagent volumes.
Claims Coverage
There are several independent claims, each outlining a distinct inventive feature central to the patent’s coverage.
Multiplex microfluidic system with parallel droplet formation and incubation
The multiplex microfluidic system comprises a chip with multiple microfluidic devices, each containing a microdroplet incubation chamber. The system includes: - An oil supply microchannel network with branches fluidically connecting an oil inlet to two or more devices, with substantially equal flow resistance. - A cell supply microchannel network with branches fluidically connecting one or more cell supply inlets to two or more microfluidic devices, also maintaining substantially equal flow resistance. - Multiple droplet formation junctions, each linked to a microfluidic device, configured to produce aqueous microdroplets suspended in oil for delivery to the incubation chamber. Each droplet formation junction is formed at an intersection between an oil supply branch and one or more cell supply branches, outputting droplets to the incubation chamber.
Method of multiplex analysis of single cell characteristics using the microfluidic system
The method includes: 1. Providing the multiplex microfluidic system, oil, at least a first cell suspension, and optionally a reagent. 2. Flowing oil and cell suspension into the respective inlets to form at least two streams of aqueous microdroplets in oil at the droplet formation junctions, with droplets entering the microdroplet incubation chambers. 3. Incubating the microdroplets to allow cell characteristic expression. 4. Analyzing single cell characteristics by observing the microdroplets during incubation.
Microdroplet sorting and analysis chip with sequential incubation and merging capabilities
This chip includes: - An oil supply microchannel from the inlet to a droplet formation junction to form aqueous microdroplets. - Cell supply microchannel(s) leading to the same junction. - A first microdroplet sorter downstream of the junction for sorting according to cell characteristics, diverting microdroplets into two or more microchannels. - A first incubation chamber downstream of the sorter. - A second sorter downstream of the first chamber for further sorting into two or more microchannels. - An optional merging junction to merge sorted microdroplets with droplets containing a reagent or additional cells, and a second incubation chamber downstream of the second sorter or merging junction.
Method of sorting and analyzing cells in microdroplets with sequential sorting, merging, and incubation
This method involves: 1. Providing the sorting and analysis chip, oil, at least a first cell suspension, and optionally a reagent or another cell suspension. 2. Forming a microdroplet stream at the droplet formation junction, with at least a portion containing one or more cells. 3. Sorting the microdroplets at the first sorter by cell characteristics, with selected droplets entering the first incubation chamber. 4. Incubating the droplets and analyzing cell characteristics during incubation. 5. Passing droplets from the first incubation chamber to the second sorter for additional sorting by cell characteristics. 6. Directing some droplets into an optional merging junction for merging with reagent or additional cell droplets before entry into the second incubation chamber. 7. Incubating and analyzing cell characteristics in the second incubation chamber.
In summary, the independent claims protect an integrated multiplex microfluidic system with matched-flow droplet formation, parallelized analysis chambers, droplet sorting and merging functions, as well as methods for complex single-cell analysis workflows and droplet handling.
Stated Advantages
Enables high-throughput, multiplexed single cell analysis and drug screening within a single integrated microfluidic chip platform.
Allows simultaneous analysis of different cells, cell combinations, or assay conditions using a single device.
Facilitates synchronous generation of uniform droplets by matching total flow resistance of microchannels.
Provides parallelization and increased throughput compared to previous systems by supporting larger droplet arrays and higher droplet filling rates.
Reduces the number of inlets and simplifies interfacing by sharing oil and analyte supplies across multiple devices.
Allows real-time monitoring and time-lapse microscopy via droplet storage arrays.
Supports enrichment and sorting of droplets containing specific cell types or phenotypes for downstream analysis.
Permits merging of droplets with reagents or additional cell types to study sequential or combinatorial treatments.
Enables the correlation of functional single-cell phenotypes with molecular/genetic analysis.
Documented Applications
High-throughput single cell drug screening applications for pharmaceutical research.
Simultaneous multi-parametric assessment of live single cell analysis and cell-cell interaction dynamics.
Screening the efficacy of cell-based immunotherapies by assessing cell function and dynamics.
Quantifying immune cell–mediated killing of tumor or microbial cells using individual or mixed cell populations.
Determining dose-response curves for drugs at the single cell level using on-board drug dilution and parallel arrays.
Profiling patient-derived cells for personalized therapy optimization and assessment.
Enrichment, manipulation, and sorting of droplets for downstream transcriptomic or genomic analysis of sorted single cells.
Assessing real-time cell behaviors such as cytokine release, cell death, and phenotypic changes in response to drugs or immune effectors.
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