Abstract: A microfluidic device includes a first substrate including at least one microfluidic channel and a plurality of microwells, as well as a cooperating second substrate defining multiple split-walled cell trap structures that are registered with and disposed within the plurality of microwells. A method for performing an assay includes flowing cells and a first aqueous medium into a plurality of microwells of a microfluidic device, wherein each microwell includes a cell trap structure configured to trap at least one cell. The method further comprises flowing a nonpolar fluid with low permeability for analytes of interest through a microfluidic channel to flush a portion of the first aqueous medium from the microfluidic channel while retaining another portion of the first aqueous medium and at least one cell within each microwell. Surface tension at a non-polar/polar medium interface prevents molecule exchange between interior and exterior portions of microwells.

Abstract: Fluorescent pH sensors are provided. The fluorescent pH sensor comprises a copolymer for sensing pH and a polymerized form of N-(2-hydroxypropyl)methacrylamide (HPMA) or 2-hydroxyethyl methacrylate (HEMA). The probe for sensing pH has formula (I): wherein R1 and R2 are as defined herein. The fluorescent pH sensors may be used in determining the pH of a sample and detecting extracellular pH in a sample. Methods for preparing the fluorescent pH sensors and the probe for sensing pH are also provided.

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: A thixotropic and biocompatible gel, formed by combining of 4-arm polyethylene glycol with fumed silica (PEG-silica). The thixotropicity of the gel is affected by the ratio of PEG to silica. To study the gels biocompatibility, the cell suspension in PBS or another medium was added to the PEG-silica gel via mixing to uniformly disperse the cells in the gel and then placed in an incubator before performing fluorescence live-dead assays. Duration of cell viability (cell life time) in gels was measured to be up to several days, depending on gel composition. Due to their optical and fluidic properties the gels are compatible with live cell imaging including 3D computed tomography and offer a means for moving the cells in a highly controllable manner by applying and removing pressure on the gels.

Abstract: Mitochondria-targeting potassium sensors and method(s) for making such sensors. The sensor shows a response to potassium and displays a 130-fold dynamic range of fluorescence intensity and high brightness. The sensors response to potassium concentrations was demonstrated to be unaffected by cellular pH value and/or concentrations of other ions. The sensors can be used for monitoring the mitochondrial potassium efflux/influx.

Abstract: A microfluidic device includes a first substrate including at least one microfluidic channel and a plurality of microwells, as well as a cooperating second substrate defining multiple split-walled cell trap structures that are registered with and disposed within the plurality of microwells. A method for performing an assay includes flowing cells and a first aqueous medium into a plurality of microwells of a microfluidic device, wherein each microwell includes a cell trap structure configured to trap at least one cell. The method further comprises flowing a nonpolar fluid with low permeability for analytes of interest through a microfluidic channel to flush a portion of the first aqueous medium from the microfluidic channel while retaining another portion of the first aqueous medium and at least one cell within each microwell. Surface tension at a non-polar/polar medium interface prevents molecule exchange between interior and exterior portions of microwells.

Abstract: The present disclosure relates to an optical luminescence dual sensor comprising a polymerized form of a probe for sensing pH; a polymerized form of a probe for sensing oxygen; a polymerized form of an internal reference probe; and a matrix. The present disclosure also relates to methods of preparing an optical luminescence dual sensor and methods of using them.

Abstract: Mitochondria-targeting potassium sensors and method(s) for making such sensors. The sensor shows a response to potassium and displays a 130-fold dynamic range of fluorescence intensity and high brightness. The sensors response to potassium concentrations was demonstrated to be unaffected by cellular pH value and/or concentrations of other ions. The sensors can be used for monitoring the mitochondrial potassium efflux/influx.

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: The present disclosure relates to an optical fluorescence sensor comprising a probe for sensing glucose, an intra-reference probe, and a matrix. The present disclosure also relates to an optical fluorescence dual sensor comprising a probe for sensing glucose having two boronic acid moieties, a probe for sensing oxygen comprising modified porphyrin, an intra-reference probe that is rhodamine-based, and a matrix. The present disclosure additionally relates to methods of preparing these sensors and methods of using them.

Abstract: A triple sensor structured for simultaneous measurement of glucose, oxygen, and pH. The sensor components are in thin film states such as sensing films or membranes, with a glucose probe associated with emission of radiation in the blue part of the spectrum, an oxygen probe associated with radiation in red portion of the spectrum, and a pH probe—with a green portion of the spectrum. The optical probes are chemically grafted or immobilized in a suitable polymer matrix, alleviating the leaching of the probes from the matrix, improving the thin film sensing stability, and enabling the repeatable use of the same sensing films.

Abstract: The present disclosure relates to an optical luminescence dual sensor comprising a polymerized form of a probe for sensing pH; a polymerized form of a probe for sensing oxygen; a polymerized form of an internal reference probe; and a matrix. The present disclosure also relates to methods of preparing an optical luminescence dual sensor and methods of using them.

Abstract: This disclosure relates generally to potassium ion sensors and monomers derived from such sensors. The disclosure also provides for polymers (e.g., random copolymers, nanoparticles, polymer thin films, and sensors) having polymerized monomeric potassium ion sensors as described herein. These compounds and polymers are useful for measuring intracellular and extracellular potassium ion concentrations.

Abstract: A triple sensor structured for simultaneous measurement of glucose, oxygen, and pH. The sensor components are in thin film states such as sensing films or membranes, with a glucose probe associated with emission of radiation in the blue part of the spectrum, an oxygen probe associated with radiation in red portion of the spectrum, and a pH probe—with a green portion of the spectrum. The optical probes are chemically grafted or immobilized in a suitable polymer matrix, alleviating the leaching of the probes from the matrix, improving the thin film sensing stability, and enabling the repeatable use of the same sensing films.

Abstract: A triple sensor structured for simultaneous measurement of glucose, oxygen, and pH. The sensor components are in thin film states such as sensing films or membranes, with a glucose probe associated with emission of radiation in the blue part of the spectrum, an oxygen probe associated with radiation in red portion of the spectrum, and a pH probe—with a green portion of the spectrum. The optical probes are chemically grafted or immobilized in a suitable polymer matrix, alleviating the leaching of the probes from the matrix, improving the thin film sensing stability, and enabling the repeatable use of the same sensing films.

Abstract: The present disclosure relates to an optical fluorescence sensor comprising a probe for sensing glucose, an intra-reference probe, and a matrix. The present disclosure also relates to an optical fluorescence dual sensor comprising a probe for sensing glucose, a probe for sensing oxygen, an intra-reference probe, and a matrix. The present disclosure additionally relates to methods of preparing these sensors and methods of using them.

Abstract: This disclosure relates generally to potassium ion sensors and monomers derived from such sensors. The disclosure also provides for polymers (e.g., random copolymers, nanoparticles, polymer thin films, and sensors) having polymerized monomeric potassium ion sensors as described herein. These compounds and polymers are useful for measuring intracellular and extracellular potassium ion concentrations.

Abstract: The present invention provides an SPR system and corresponding methods of use, for determining the presence or concentration of tumor-associated antigens in cancer patient samples. The SPR system may have multiple channels, with each channel having operably affixed thereto an antibody specific for a tumor-associated antigen, so as to allow detection of multiple tumor-associated antigens simultaneously. When a biological sample from a patient is applied to the SPR system, the presence of two or more tumor-associated antigens can be determined by measuring an SPR signal shift from each channel. The SPR system may detect the presence or concentration of a tumor-associated carbohydrate antigen, where the sensor surface contains affixed thereto an antibody specific for the glycosyl epitope, as well as an antibody specific for the polypeptide to which the carbohydrate antigen is naturally associated in cancer patients.