Single cell isolation and processing system with reversible well shape
Inventors
KONRY, Tania • UGOLINI, Giovanni
Assignees
Northeastern University Boston
Publication Number
US-12064768-B2
Publication Date
2024-08-20
Expiration Date
2041-02-22
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Abstract
The technology provides arrays of microwells with reversible shape for culture, analysis, and recovery of individual cells or groups of cells. The microwells are reversibly formed by vacuum-induced deflection of an elastomeric membrane into an array of microwell molds formed in a microwell substrate. The shape of each microwell or groups of microwells can be selectively altered by applying or releasing vacuum to individual microwells. The devices, systems, and methods utilizing the technology enable collection of individual cells for further study or therapeutic use without the need for micromanipulation.
Core Innovation
The present technology provides arrays of microwells with variable, reversible shape for culture, analysis, and recovery of individual cells or groups of cells. Each microwell is formed by applying vacuum beneath a thin, flexible elastomeric membrane covering an array of microwell molds within a solid substrate. By selectively applying or releasing vacuum via pneumatic microchannels, the membrane conformably forms microwells or returns to a planar state above the molds.
This system is designed to address complications associated with conventional microwell arrays that are monolithic and permanently formed, requiring complex, costly, and low-throughput micromanipulation for cell recovery. The technology simplifies the isolation and cultivation of individual cells after cultivation in microwells, eliminating the operational difficulties, expense, and delays of traditional methods.
The reversible microwell design allows for targeted release and recovery of specific cells without disturbing unselected wells, enabling collection for further analysis or therapeutic use without micromanipulation. The microwell array, with its adjustable configuration, may be automated and integrated with imaging and perfusion systems, enabling high-throughput, precise, and programmable single-cell manipulation and recovery.
Claims Coverage
The patent includes one independent claim, which defines several key inventive features.
Reversible microwell device with elastomeric membrane and pneumatic microchannels
A microfluidic device featuring: - A microwell substrate with a plurality of microwell molds, each having an opening at the substrate's upper surface. - One or more pneumatic microchannels connecting each microwell mold to an inlet for attaching a controllable vacuum source. - An elastomeric membrane bonded to the upper substrate surface and covering the microwell mold openings. When vacuum is applied via a connected pneumatic microchannel, the elastomeric membrane conformably forms a microwell; when pressure returns to atmospheric, the membrane relaxes to a planar surface above each microwell mold.
System for single cell analysis and isolation with programmable vacuum control and cell analysis modules
A system comprising: - The above microfluidic device. - A controllable vacuum source connectable to the pneumatic microchannels and operative to deform the elastomeric membrane. - A microscope configured for observing cells in microwells. - A controller for programmed operation of the controllable vacuum source and/or directing vacuum to selected pneumatic channels. The system enables both the mechanical operation and observation/analysis of cells within reversible microwells.
Method for analysis and isolation of single cells using reversible microwell system
A method comprising: 1. Providing the system described above. 2. Applying vacuum to one or more microwell molds to form microwells by deforming the elastomeric membrane. 3. Depositing a cell suspension onto the device so cells become entrapped within microwells. 4. Analyzing entrapped cells with the microscope to determine cell characteristics. 5. Isolating cells having a characteristic of interest as determined by analysis. This method enables cell analysis and selective isolation based on observed characteristics, using the reversible microwell device and system.
The inventive features provide a reversible microwell substrate employing pneumatic actuation to form and release microwells, combined with system elements for programmable control and analysis, and methods for cell analysis and isolation utilizing these components.
Stated Advantages
Provides facile and uncomplicated techniques for isolation and cultivation of individual cells after cultivation in microwells.
Eliminates or reduces the need for costly, complex, and low-throughput micromanipulation and cell sorting techniques.
Enables accurate and quick isolation of single cells or groups of cells with potential for rapid scale-up and automation.
Allows selective recovery of desired cells after assays or culture without disturbing unselected cells.
Facilitates integration with laboratory automation, imaging, software, and high-throughput cell analysis systems.
Reusable devices allowing for multiple cell screenings.
Documented Applications
Single cell analyses for immunotherapy, single cell proteomics, and rapid screening.
Analysis of tumor cell interactions with on-demand release and collection of functionally identified cells.
Screening and isolation of cells based on secretion of biomolecules at the single cell level.
Recovery and cultivation of drug-resistant cancer cells after chemotherapy exposure for further study.
Correlating single cell functional data (e.g., secretomic or functional assay results) with genotype or phenotype.
Formation and analysis of multicellular aggregates such as tumor groups, neurospheres, mammospheres, hepatospheres, and embryoid bodies.
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