Microfluidic devices and methods based on massively parallel picoreactors for cell and molecular diagnostics
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Publication Number
US-9110026-B2
Publication Date
2015-08-18
Expiration Date
Abstract
Microfluidic devices and methods of forming cell reactors for performing cell analysis in a microfluidic chip. A microfluidic chip, in one implementation, includes a plurality of trapping sites, each of the plurality of trapping sites having a plurality of micropillars configured to trap one or more cells in an interior space formed by the plurality of micropillars. The plurality of micropillars in each trapping site form a picoreactor for cell and molecular diagnostics, such as characterizing, isolation, processing, and amplification of different cells, cells containing different substances, different particles, different biochemical compositions, proteins, and enzymes.
Core Innovation
The invention provides a method of forming cell reactors for performing cell analysis in a microfluidic chip. One or more cells are trapped in at least one of a plurality of trapping sites, and each trapping site has a plurality of micropillars configured to trap the one or more cells in an interior space formed by the plurality of micropillars. After trapping, an encapsulated reactor array is formed for the trapped cells in the microfluidic chip.
The method encapsulates the trapped cells by flowing a first immiscible fluid around the one or more cells at the plurality of trapping sites. This encapsulation is performed after trapping of the one or more cells, resulting in an encapsulated reactor array in the microfluidic chip. The trapped-and-encapsulated architecture is described as a microfluidic massively parallel picoreactor platform for cell and molecular diagnostics.
In further described implementations, the platform enables integrated multistep processing in the encapsulated reactor array, including reverse transcription and RT-PCR/qPCR with on-chip thermal cycling using hot immiscible fluid and cold immiscible fluid circulating from respective hot and cold reservoirs. Additional described processing includes in-situ electrical processing of molecules or genes obtained from one or more cells, and downstream electrophoresis with EWOD-based droplet movement. The document also describes additional sample preprocessing and mRNA purification based on magnetic-bead droplet fission/fusion.
Claims Coverage
The independent claim covers forming an encapsulated reactor array in a microfluidic chip by micropillar-based trapping followed by encapsulation with an immiscible fluid. Across the dependent claims, multiple inventive features refine the micropillar trapping structure and extend the workflow to thermal cycling, in-situ electrical processing, and aligned reagent microarray spotting.
Micropillar-Based Trapping at Plurality of Trapping Sites
Trapping one or more cells in at least one of a plurality of trapping sites, where each trapping site includes a plurality of micropillars configured to trap the one or more cells in an interior space formed by the plurality of micropillars.
Immiscible-Fluid Encapsulation Forming Encapsulated Reactor Array
Flowing a first immiscible fluid around the one or more cells to encapsulate the one or more cells at the plurality of trapping sites to form an encapsulated reactor array, after the trapping of the one or more cells.
Inner and Outer Micropillar Layer Configuration
Arranging a plurality of micropillars as an inner layer to trap one or more cells and as an outer layer to keep the first immiscible fluid surrounding the trapped cells.
Thermal Cycling with Hot and Cold Immiscible Fluids
Circulating a hot immiscible fluid and a cold immiscible fluid from respective hot and cold reservoirs to carry out thermal cycling.
In-Situ Electrical Processing of Molecules or Genes
Performing in-situ electrical processing of molecules or genes obtained from one or more cells.
Microarray Reagent Spotting Aligned to Trapping Sites
Spotting reagents onto a microarray on the bottom portion of a microfluidic chip, with the spotted reagent locations aligned to a plurality of trapping sites on the top portion of the chip.
Overall claim coverage centers on micropillar configured trapping sites followed by immiscible-fluid encapsulation to form an encapsulated reactor array, with refinements including inner/outer micropillar layer function, thermal cycling using hot and cold immiscible fluids, in-situ electrical processing of molecules or genes, and reagent microarray spotting aligned to trapping-site locations.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Cell and molecular diagnostics using a microfluidic massively parallel picoreactor platform.
Prenatal DMD diagnosis using described diagnostic examples.
Cancer diagnostics including CTC copy-number and SC-PCR as described examples.
HCV analysis in PBMCs as described examples.
HIV/AIDS analysis as described examples.
Biomedical research including use with iPS cells as described examples.
Combinatorial drug screening using a Programmable Array of Living (PAL) cells, including paired drug concentration and combinations in isolated reactor compartments as described examples.
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