Abstract: Systems and methods of using the same for functional fluorescence imaging of live cells in suspension with isotropic three dimensional (3D) diffraction-limited spatial resolution are disclosed. The method-live cell computed tomography (LCCT)-involves the acquisition of a series of two dimensional (2D) pseudo-projection images from different perspectives of the cell that rotates around an axis that is perpendicular to the optical axis of the imaging system. The volumetric image of the cell is then tomographically reconstructed.

Abstract: Systems and methods of using the same for functional fluorescence imaging of live cells in suspension with isotropic three dimensional (3D) diffraction-limited spatial resolution are disclosed. The method-live cell computed tomography (LCCT)-in-volves the acquisition of a series of two dimensional (2D) pseudo-projection images from different perspectives of the cell that rotates around an axis that is perpendicular to the optical axis of the imaging system. The volumetric image of the cell is then tomographically reconstructed.

Abstract: An integrated technological platform enabling real-time quantitative multiparameter metabolic profiling, utilizing either or both of extra and intracellular optical sensors, individually or simultaneously. A scalable embedded micropocket array structure, generally fabricated on fused silica substrates, facilitates the integration of multiple, spatially separated extracellular sensors for multiparameter analysis in a container formed with the use of an activation mechanism forming part of a device configured to hold the container during the measurements. The creation of hermetically sealed microchambers is carried out with a pneumatically and/or mechanically and/or electromechanically driven device that is “floating” within the holding device and that is optionally equipped with a vacuum/suction mechanism to hold a component of the container at its surface.

Abstract: A device for high-throughput multi-parameter functional profiling of the same cells in multicellular settings and in isolation is provided. In certain aspects, an integrated microfluidic device for multi-parameter metabolic and other phenotypic profiling of live biological cells is useable with: 1) multicellular clusters or small biopsy tissue samples, 2) cultures of the constituent cells obtained after cluster/tissue dissociation, and 3) the same constituent single cells in isolation. The approach enables study of the effects of multicellular complexity, such as in response to treatment, pathogens, stress, or other factors concerning disease origination and progression. Measurements may be performed on single cells or multicellular populations or tissues in the same assay at the same time.

Abstract: An integrated technological platform enabling real-time quantitative multiparameter metabolic profiling, utilizing either or both of extra and intracellular optical sensors, individually or simultaneously. A scalable embedded micropocket array structure, generally fabricated on fused silica substrates, facilitates the integration of multiple, spatially separated extracellular sensors for multiparameter analysis in a container formed with the use of an activation mechanism forming part of a device configured to hold the container during the measurements. The creation of hermetically sealed microchambers is carried out with a pneumatically and/or mechanically and/or electromechanically driven device that is “floating” within the holding device and that is optionally equipped with a vacuum/suction mechanism to hold a component of the container at its surface.

Abstract: Systems and methods of using the same for functional fluorescence imaging of live cells in suspension with isotropic three dimensional (3D) diffraction-limited spatial resolution are disclosed. The method-live cell computed tomography (LCCT)-in-volves the acquisition of a series of two dimensional (2D) pseudo-projection images from different perspectives of the cell that rotates around an axis that is perpendicular to the optical axis of the imaging system. The volumetric image of the cell is then tomographically reconstructed.

Abstract: Systems and methods of using the same for functional fluorescence imaging of live cells in suspension with isotropic three dimensional (3D) diffraction-limited spatial resolution are disclosed. The method-live cell computed tomography (LCCT)-involves the acquisition of a series of two dimensional (2D) pseudo-projection images from different perspectives of the cell that rotates around an axis that is perpendicular to the optical axis of the imaging system. The volumetric image of the cell is then tomographically reconstructed.

Abstract: The invention relates to microfluidic devices and array disks for “one-pot” isolated chemical reactions. The array disks comprise a plurality of sectors in which each sector comprises one microfluidic device. The microfluidic devices comprise a fluid delivery channel and an array of wells wherein the fluid delivery channel delivery fluid into the wells in a serpentine arrangement. In some embodiments, the fluid delivery channel is directly above the array of wells. In other embodiments, the fluid delivery channel is offset from the array of wells so that side channels branching from the fluid delivery channel delivers fluid into the wells. The well of the microfluidic device comprises a gas-permeable membrane that forms the floor, well, or at least a portion of the floor or wall of the well. In preferred embodiments, the well is cylindrical.

Abstract: A device for high-throughput multi-parameter functional profiling of the same cells in multicellular settings and in isolation is provided. In certain aspects, an integrated microfluidic device for multi-parameter metabolic and other phenotypic profiling of live biological cells is useable with: 1) multicellular clusters or small biopsy tissue samples, 2) cultures of the constituent cells obtained after cluster/tissue dissociation, and 3) the same constituent single cells in isolation. The approach enables study of the effects of multicellular complexity, such as in response to treatment, pathogens, stress, or other factors concerning disease origination and progression. Measurements may be performed on single cells or multicellular populations or tissues in the same assay at the same time.

Abstract: Systems and methods for in situ laser lysis for analysis of biological tissue (live, fixed, frozen or otherwise preserved) at single cell resolution in 3D. For example, a system and method for lysing individual cells in situ, including the steps of capturing a tissue sample comprising a cellular content, subjecting the tissue sample to a stream of continuous fluid flow, lysing a selected area of the tissue sample with a laser, thereby releasing at least a portion of the cellular content from the tissue sample, recovering at least one target molecule from the cellular content in the stream, and processing at least one target molecule is provided. The system collects cellular contents, performs highly multiplexed (RT-qPCR or RNA-seq), and sequentially (cell-by-cell) reconstructs a 3D spatial map of mRNA expression of the tissue with a large number of genes. A 3D spatial map of the DNA, RNA, and/or proteins can be generated for each cell in the tissue.

Abstract: The invention relates to microfluidic devices and array disks for “one-pot” isolated chemical reactions. The array disks comprise a plurality of sectors in which each sector comprises one microfluidic device. The microfluidic devices comprise a fluid delivery channel and an array of wells wherein the fluid delivery channel delivery fluid into the wells in a serpentine arrangement. In some embodiments, the fluid delivery channel is directly above the array of wells. In other embodiments, the fluid delivery channel is offset from the array of wells so that side channels branching from the fluid delivery channel delivers fluid into the wells. The well of the microfluidic device comprises a gas-permeable membrane that forms the floor, well, or at least a portion of the floor or wall of the well. In preferred embodiments, the well is cylindrical.

Abstract: The invention relates to microfluidic devices and array disks for “one-pot” isolated chemical reactions. The array disks comprise a plurality of sectors in which each sector comprises one microfluidic device. The microfluidic devices comprise a fluid delivery channel and an array of wells wherein the fluid delivery channel delivery fluid into the wells in a serpentine arrangement. In some embodiments, the fluid delivery channel is directly above the array of wells. In other embodiments, the fluid delivery channel is offset from the array of wells so that side channels branching from the fluid delivery channel delivers fluid into the wells. The well of the microfluidic device comprises a gas-permeable membrane that forms the floor, well, or at least a portion of the floor or wall of the well. In preferred embodiments, the well is cylindrical.

Abstract: Novel methods and devices including modified endoscopes that employ fiber optic technology and multiple extrinsic optical sensors for enablement of simultaneous determination of change rates of biological analytes in vivo with single-cell resolution. The devices employ dynamic oscillatory actuation to determine the gradient of analytes of interest along one or multiple directions with respect to the cell. The change rate or flux of the analyte of interest can be easily determined by applying the first Fick's law that relates the flux with the concentration gradient. The dual operation mode of the device markedly increases measurement sensitivity and accuracy.

Abstract: Systems and methods for in situ laser lysis for analysis of biological tissue (live, fixed, frozen or otherwise preserved) at single cell resolution in 3D. For example, a system and method for lysing individual cells in situ, including the steps of capturing a tissue sample comprising a cellular content, subjecting the tissue sample to a stream of continuous fluid flow, lysing a selected area of the tissue sample with a laser, thereby releasing at least a portion of the cellular content from the tissue sample, recovering at least one target molecule from the cellular content in the stream, and processing at least one target molecule is provided. The system collects cellular contents, performs highly multiplexed (RT-qPCR or RNA-seq), and sequentially (cell-by-cell) reconstructs a 3D spatial map of mRNA expression of the tissue with a large number of genes. A 3D spatial map of the DNA, RNA, and/or proteins can be generated for each cell in the tissue.

Abstract: The present invention provides a system and method for lysing individual cells in situ, including the steps of capturing a tissue sample comprising a cellular content, subjecting the tissue sample to a stream of continuous fluid flow, lysing a selected area of the tissue sample with a laser, thereby releasing at least a portion of the cellular content from the tissue sample, recovering at least one target molecule from the cellular content in the stream, and processing the at least one target molecule.

Abstract: The invention relates to microfluidic devices and array disks for “one-pot” isolated chemical reactions. The array disks comprise a plurality of sectors in which each sector comprises one microfluidic device. The microfluidic devices comprise a fluid delivery channel and an array of wells wherein the fluid delivery channel delivery fluid into the wells in a serpentine arrangement. In some embodiments, the fluid delivery channel is directly above the array of wells. In other embodiments, the fluid delivery channel is offset from the array of wells so that side channels branching from the fluid delivery channel delivers fluid into the wells. The well of the microfluidic device comprises a gas-permeable membrane that forms the floor, well, or at least a portion of the floor or wall of the well. In preferred embodiments, the well is cylindrical.

Abstract: Novel methods and devices including modified endoscopes that employ fiber optic technology and multiple extrinsic optical sensors for enablement of simultaneous determination of change rates of biological analytes in vivo with single-cell resolution. The devices employ dynamic oscillatory actuation to determine the gradient of analytes of interest along one or multiple directions with respect to the cell. The change rate or flux of the analyte of interest can be easily determined by applying the first Fick's law that relates the flux with the concentration gradient. The dual operation mode of the device markedly increases measurement sensitivity and accuracy.