Abstract: A microfluidic diagnostic device with a three-dimensional (3D) flow architecture comprises a polymeric body having first and second opposing surfaces and comprising first flow channels in the first opposing surface, second flow channels in the second opposing surface, and connecting flow passages extending through a thickness of the polymeric body to connect the first flow channels to the second flow channels, thereby defining a continuous 3D flow pathway in the polymeric body. The microfluidic diagnostic device also includes a first cover adhered to the first opposing surface to seal the first flow channels, a second cover adhered to the second opposing surface to seal the second flow channels, and one or more access ports in fluid communication with the continuous 3D flow pathway for introducing liquid reagent(s) and/or a sample into the polymeric body.

Abstract: Provided are methods and related devices for preparing a cell and tissue culture, including a hanging drop culture. Microwells are specially loaded with cell mixtures using a removable reservoir and forcing cells into the underlying microwells. The removable reservoir is removed and the cells partitioned into the individual microwells and covered by an immiscible layer of fluid. The microwells and immiscible layer is inverted and the cells in the microwells cultured. The microwells may have shape-controlling elements to control the three-dimensional shape of the culture.

Abstract: A stacked testing assembly (100) includes a microfluidic cartridge (10) for analyzing a fluid sample and a testing setup, said microfluidic cartridge includes a number of layers (1, 2) stacked in a height direction with many different kinds of combinations, said testing setup (20) is capable of assembling and testing all kinds of said layers combinations with no change to the setup.

Abstract: The present disclosure provides a biological machine comprising a skeleton comprising at least one hydrogel strip and at least two hydrogel bases, and at least one bioactuator, the at least one bioactuator comprising at least one muscle ring, the muscle ring comprising skeletal muscle cells, wherein the muscle ring is tethered around the at least two pillars. The muscle ring can comprises skeletal muscle, and skeletal muscle innervated with motor neurons, allowing chemical control of muscle contraction. Methods of making the biological machine is also provided.

Abstract: Various methods and devices for spatial molecular analysis from tissue is provided. For example, a method of spatially mapping a tissue sample is provided with a microarray having a plurality of wells, wherein adjacent wells are separated by a shearing surface; overlaying said microarray with a tissue sample; applying a deformable substrate to an upper surface of said tissue sample; applying a force to the deformable substrate, thereby forcing underlying tissue sample into the plurality of wells; shearing the tissue sample along the shearing surface into a plurality of tissue sample islands, with each unique tissue sample island positioned in a unique well; and imaging or quantifying said plurality of tissue sample islands, thereby generating a spatial map of said tissue sample. The imaging and/or quantifying may use a nucleic acid amplification technique.

Abstract: Provided herein are multiplexible particle systems and related methods of making and using the multiplexible particle systems. A plurality of monodisperse polymer particle populations are provided, wherein each population has a unique electrical parameter for multiplexed detection by flow through a spatially confined electric field, and the distribution of the electrical parameter within each population is sufficiently narrow for reliable multiplex detection. The density difference between populations may be relatively uniform, such as within 30%, including within 30% of a suspending solution density for when the particles are flowed through a confined electric field and detected in a multiplex manner by a change in the electric parameter measured by a counting device. Relatively uniform density of particles is important for ensuring minimal settling while the plurality of particle populations flow together under a single flow regime.

Abstract: Described herein are systems and methods which utilize an array of wells to isolate pathogens and nucleic acid detection techniques to accurately and rapidly detect pathogens in fluid samples, even in very low concentrations, including from solid or semi-solid samples that have been fluidized. The provided systems and methods dry the fluid sample to deposit a fraction of the total volume in a number of wells and perform nucleic acid detection on individual wells to detect even individual pathogens and provide a quantitative analysis of the amount of pathogen within the sample. Also provided are methods and systems for precise delivery of dried materials, including biomolecules that are enzymes of use in the process, to microwells.

Abstract: Various methods and devices for spatial molecular analysis from tissue is provided. For example, a method of spatially mapping a tissue sample is provided with a microarray having a plurality of wells, wherein adjacent wells are separated by a shearing surface; overlaying said microarray with a tissue sample; applying a deformable substrate to an upper surface of said tissue sample; applying a force to the deformable substrate, thereby forcing underlying tissue sample into the plurality of wells; shearing the tissue sample along the shearing surface into a plurality of tissue sample islands, with each unique tissue sample island positioned in a unique well; and imaging or quantifying said plurality of tissue sample islands, thereby generating a spatial map of said tissue sample. The imaging and/or quantifying may use a nucleic acid amplification technique.

Abstract: Provided are methods and devices for the label free detection of analytes in solution, including analytes suspended in a biological fluid. A field effect transistor (FET) is positioned in close proximity to a paired set of reference electrodes and the reference electrodes electrically biased to provide desalting and a stable gate voltage to the FET. In this manner, charged ions are depleted in the sensing region of the sensor and device sensitivity to analyte detection improved by the removal of charge that otherwise interferes with measurement. Also provided are methods and systems providing increased in reference electrode surface area and/or decrease in droplet volume to further improve label-free detection of analytes.

Abstract: A sample carrier may include a sample preparation module and an amplification module. A sample mixes with a lysis medium and a nucleic acid amplification medium in the sample preparation module and then flows into a plurality of microfluidic chambers in the amplification module. The microfluidic chambers have disposed therein primers configured to initiate amplification of one or more target nucleic acid sequences corresponding to one or more pathogens. The sample carrier is inserted into an apparatus that includes a plurality of Sight sources and a camera. The light sources illuminate the microfluidic chambers with excitation light, a fluorophore emits fluorescence light indicative of nucleic acid amplification in response to the excitation-light, and the camera captures images of the microfluidic chambers. A target nucleic acid sequence in the sample is indicated by the images showing an increasing fluorescence in a microfluidic chamber that has the primers for that sequence.

Abstract: This disclosure relates to methods and devices to count particles of interest, such as cells. The methods include obtaining a fluid sample that may contain particles of interest; counting all types of particles in a portion of the sample using a first electrical differential counter to generate a first total; removing any particles of interest from the portion of the fluid sample; counting any particles remaining in the portion of the fluid sample using a second electrical differential counter after the particles of interest are removed to generate a second total; and calculating a number of particles of interest originally in the fluid sample by subtracting the second total from the first total, wherein the difference is the number of particles of interest in the sample. These methods and related devices can be used, for example, to produce a robust, inexpensive diagnostic kit for CD4+ T cell counting in whole blood samples.

Abstract: Provided are methods and devices for the label free detection of analytes in solution, including analytes suspended in a biological fluid. A field effect transistor (FET) is positioned in close proximity to a paired set of reference electrodes and the reference electrodes electrically biased to provide desalting and a stable gate voltage to the FET. In this manner, charged ions are depleted in the sensing region of the sensor and device sensitivity to analyte detection improved by the removal of charge that otherwise interferes with measurement. Also provided are methods and systems providing increased in reference electrode surface area and/or decrease in droplet volume to further improve label-free detection of analytes.

Abstract: Provided herein are methods and devices for characterizing a biomolecule parameter by a nanopore-containing membrane, and also methods for making devices that can be used in the methods and devices provided herein. The nanopore membrane is a multilayer stack of conducting layers and dielectric layers, wherein an embedded conducting layer or conducting layer gates provides well-controlled and measurable electric fields in and around the nanopore through which the biomolecule translocates. In an aspect, the conducting layer is graphene.

Abstract: Provided are methods and systems that characterize a property of a particle suspended in a fluid sample in a label-free manner. Detection elements are provided fluidically adjacent upstream and downstream from a modulation element. Fluid sample containing particles flows across a first detection element and a first particle parameter detected for each particle that passes the first detection element or a first aggregate particle parameter for a plurality of particles that pass the first detection element. The particles flow from the first detection element to a first modulation element, wherein the first modulation element effects a change in a property of the particles flowing past the first modulation element. A second detection element then detects the particle parameter again or a second aggregate particle parameter for a plurality of particles that pass the second detection element. Comparing the first and second particle or aggregate parameters thereby characterizes the particle property.

Abstract: Provided herein are methods and devices for characterizing a biomolecule parameter by a nanopore-containing membrane, and also methods for making devices that can be used in the methods and devices provided herein. The nanopore membrane is a multilayer stack of conducting layers and dielectric layers, wherein an embedded conducting layer or conducting layer gates provides well-controlled and measurable electric fields in and around the nanopore through which the biomolecule translocates. In an aspect, the conducting layer is graphene.

Abstract: This disclosure relates to methods and devices to count particles of interest, such as cells. The methods include obtaining a fluid sample that may contain particles of interest; counting all types of particles in a portion of the sample using a first electrical differential counter to generate a first total; removing any particles of interest from the portion of the fluid sample; counting any particles remaining in the portion of the fluid sample using a second electrical differential counter after the particles of interest are removed to generate a second total; and calculating a number of particles of interest originally in the fluid sample by subtracting the second total from the first total, wherein the difference is the number of particles of interest in the sample. These methods and related devices can be used, for example, to produce a robust, inexpensive diagnostic kit for CD4+ T cell counting in whole blood samples.

Abstract: The invention provides locomotive biological machines comprised of hydrogel structures and one or more types of cells. The locomotive biological machines are capable of controlled directional movement and can be used for sensing, information processing, actuation, protein expression, and transportation.

Abstract: The present disclosure provides a bio-field effect transistor (BioFET) and a method of fabricating a BioFET device. The method includes forming a BioFET using one or more process steps compatible with or typical to a complementary metal-oxide-semiconductor (CMOS) process. The BioFET device may include a substrate; a gate structure disposed on a first surface of the substrate and an interface layer formed on the second surface of the substrate. The interface layer may allow for a receptor to be placed on the interface layer to detect the presence of a biomolecule or bio-entity. An amplification factor of the BioFET device may be provided by a difference in capacitances associated with the gate structure on the first surface and with the interface layer formed on the second surface.

Abstract: A stacked testing assembly (100) includes a microfluidic cartridge (10) for analyzing a fluid sample and a testing setup, said microfluidic cartridge includes a number of layers (1, 2) stacked in a height direction with many different kinds of combinations, said testing setup (20) is capable of assembling and testing all kinds of said layers combinations with no change to the setup.

Abstract: This disclosure relates to methods and devices to count particles of interest, such as cells. The methods include obtaining a fluid sample that may contain particles of interest; counting all types of particles in a portion of the sample using a first electrical differential counter to generate a first total; removing any particles of interest from the portion of the fluid sample; counting any particles remaining in the portion of the fluid sample using a second electrical differential counter after the particles of interest are removed to generate a second total; and calculating a number of particles of interest originally in the fluid sample by subtracting the second total from the first total, wherein the difference is the number of particles of interest in the sample. These methods and related devices can be used, for example, to produce a robust, inexpensive diagnostic kit for CD4+ T cell counting in whole blood samples.