Abstract: The present invention provides a microscope slide for analyzing a tissue sample. The slide includes an elongate planar substrate, which comprises a first major surface having a sample-affixing area within which a tissue sample may be affixed; a first positive control sample affixed on the first major surface at a first location adjacent to the sample-affixing area; and a second positive control sample affixed on the first major surface at a second location adjacent to the sample-affixing area. The first and second locations are spaced such that quality of the staining of the first and second positive control samples is indicative of the quality of the staining of the tissue sample. Also provided are a kit containing the microscope slide and a method of making and a method of using the microscope slide.

Abstract: The present disclosure relates to characterization of biological samples. By way of example, a biological sample may be contacted with a plurality of probes specific for targets in the sample, such as probes for immune markers and segmenting probes. Acquired image data of the sample may be used to segment the images into epithelial and stromal regions to characterize individual cells in the sample based on the binding of the probes. Further, the biological sample may be characterized by a distribution, location, and type of a plurality of the characterized cells.

Abstract: The present invention provides a microscope slide for analyzing a tissue sample. The slide includes an elongate planar substrate, which comprises a first major surface having a sample-affixing area within which a tissue sample may be affixed; a first positive control sample affixed on the first major surface at a first location adjacent to the sample-affixing area; and a second positive control sample affixed on the first major surface at a second location adjacent to the sample-affixing area. The first and second locations are spaced such that quality of the staining of the first and second positive control samples is indicative of the quality of the staining of the tissue sample. Also provided are a kit containing the microscope slide and a method of making and a method of using the microscope slide.

Abstract: The present disclosure relates to characterization of biological samples. By way of example, a biological sample may be contacted with a plurality of probes specific for targets in the sample, such as probes for immune markers and segmenting probes. Acquired image data of the sample may be used to segment the images into epithelial and stromal regions to characterize individual cells in the sample based on the binding of the probes. Further, the biological sample may be characterized by a distribution, location, and type of a plurality of the characterized cells.

Abstract: Improved systems and methods for the analysis of digital images are provided. More particularly, the present disclosure provides for improved systems and methods for the analysis of digital images of biological tissue samples. Exemplary embodiments provide for: i) segmenting, ii) grouping, and iii) quantifying molecular protein profiles of individual cells in terms of sub cellular compartments (nuclei, membrane, and cytoplasm). The systems and methods of the present disclosure advantageously perform tissue segmentation at the sub-cellular level to facilitate analyzing, grouping and quantifying protein expression profiles of tissue in tissue sections globally and/or locally. Performing local-global tissue analysis and protein quantification advantageously enables correlation of spatial and molecular configuration of cells with molecular information of different types of cancer.

Abstract: Improved systems and methods for the analysis of digital images are provided. More particularly, the present disclosure provides for improved systems and methods for the analysis of digital images of biological tissue samples. Exemplary embodiments provide for: i) segmenting, ii) grouping, and iii) quantifying molecular protein profiles of individual cells in terms of sub cellular compartments (nuclei, membrane, and cytoplasm). The systems and methods of the present disclosure advantageously perform tissue segmentation at the sub-cellular level to facilitate analyzing, grouping and quantifying protein expression profiles of tissue in tissue sections globally and/or locally. Performing local-global tissue analysis and protein quantification advantageously enables correlation of spatial and molecular configuration of cells with molecular information of different types of cancer.

Abstract: The invention relates generally to a process of analyzing and visualizing the expression of biomarkers in an individual cell wherein the cell is examined to develop patterns of expression by using a grouping algorithm, and a system to perform and display the analysis.

Abstract: A method is provided for scaling an image taken at a given exposure time to a selected exposure time. The method generally comprises: determining a dark pixel intensity of an imaging device; acquiring a first image at a given exposure time; and adjusting a pixel intensity of one or more pixels in the first image, based at least in part on the dark pixel intensity, for a second exposure time that is different from the given exposure time.

Abstract: Systems and methods are disclosed for selecting a field of view for a biological sample, selecting first and second biomarkers, each having a corresponding image in a multiplexed biomarker image of the selected field of view, automatically analyzing expression level criterion for each of the first and second biomarkers, rendering the corresponding images of the selected field-of-view in an overlaid manner on the graphical user interface and highlighting a first set of biological units in the biological tissue that meets both the biomarker expression level criterion for both the first and second biomarkers.

Abstract: Methods for probing multiple targets in a biological sample are provided. The methods include the steps of providing a sample containing multiple targets, binding at least one probe having a binder coupled to an enzyme to one or more target present in the sample, and reacting the bound probe with an enzyme substrate coupled to a fluorescent signal generator. The methods include the steps of observing a signal from the fluorescent signal generator and applying to the sample a solution containing an oxidizing agent that substantially inactivates both the fluorescent signal generator and the enzyme. The methods further include the steps of binding at least one probe having a binder coupled to an enzyme to one or more target present in the sample of step, reacting the bound probe with an enzyme substrate coupled to a fluorescent signal generator; and observing a signal from the fluorescent signal generator.

Abstract: Methods for probing multiple targets in a biological sample are provided. The methods include the steps of providing a sample containing multiple targets, binding at least one probe having a binder coupled to an enzyme to one or more target present in the sample, and reacting the bound probe with an enzyme substrate coupled to a fluorescent signal generator. The methods include the steps of observing a signal from the fluorescent signal generator and applying to the sample a solution containing an oxidizing agent that substantially inactivates both the fluorescent signal generator and the enzyme. The methods further include the steps of binding at least one probe having a binder coupled to an enzyme to one or more target present in the sample of step, reacting the bound probe with an enzyme substrate coupled to a fluorescent signal generator; and observing a signal from the fluorescent signal generator.

Abstract: Methods for scaling an image taken an optimal exposure time to a selected exposure time, generally comprising, determining a dark pixel intensity of an imaging device; acquiring a first image at an optimal exposure time; and adjusting a pixel intensity of one or more pixels in the first image, based at least in part on the dark pixel intensity, for a second exposure time that is different from the optimal exposure time.

Abstract: The present application discloses a technique for obtaining and storing data on expression of multiple biomarkers in individual cells or the compartments of individual cells in a tissue specimen and methods of utilizing that data to create groups, the members of which share some degree of similarity greater than the general population from which the data is drawn, by an analysis of digital images of a portion of the tissue specimen which has been iteratively stained to generate optical signals, typically fluorescent, reflective of the amount of each of the biomarkers examined.

Abstract: Methods for probing multiple targets in a biological sample are provided. The methods include the steps of providing a biological sample containing multiple targets adhered to a solid support, binding at least one fluorescent probe to one or more target present in the sample, and observing a signal from the fluorescent probe. The method further includes the steps of oxidizing the bound fluorescent probe with a solution containing an oxidizing agent that substantially inactivates the fluorescent probe, binding at least one fluorescent probe to one or more target present in the sample, and observing a signal from the fluorescent probe. The methods disclosed herein also provide for multiple iterations of binding, observing, and oxidizing for deriving information about multiple targets in a single sample. An associated kit is also provided.

Abstract: Methods for probing multiple targets in a biological sample are provided. The methods include the steps of providing a biological sample containing multiple targets, binding at least one fluorescent probe to one or more target present in the sample, and binding at least at least one control probe to one or more target present in the sample. The methods include the steps of observing a signal from the fluorescent probe and a control signal from the control probe and applying to the sample a basic solution containing an oxidizing agent that selectively inactivates the fluorescent probe and not the control probe. The methods further include the steps of binding at least one fluorescent probe to one or more target present in the sample and observing a signal from the fluorescent probe. The methods disclosed herein also provide for multiple iterations of binding, observing, and oxidizing for deriving information about multiple targets in a single sample. An associated kit is also provided.

Abstract: Methods for probing multiple targets in a biological sample are provided. The methods include the steps of providing a biological sample containing multiple targets adhered to a solid support, binding at least one fluorescent probe to one or more target present in the sample, and observing a signal from the fluorescent probe. The method further includes the steps of oxidizing the bound fluorescent probe with a solution containing an oxidizing agent that substantially inactivates the fluorescent probe, binding at least one fluorescent probe to one or more target present in the sample, and observing a signal from the fluorescent probe. The methods disclosed herein also provide for multiple iterations of binding, observing, and oxidizing for deriving information about multiple targets in a single sample. An associated kit is also provided.

Abstract: Methods for probing multiple targets in a biological sample are provided. The methods include the steps of providing a biological sample containing multiple targets, binding at least one fluorescent probe to one or more target present in the sample, and binding at least at least one control probe to one or more target present in the sample. The methods include the steps of observing a signal from the fluorescent probe and a control signal from the control probe and applying to the sample a basic solution containing an oxidizing agent that selectively inactivates the fluorescent probe and not the control probe. The methods further include the steps of binding at least one fluorescent probe to one or more target present in the sample and observing a signal from the fluorescent probe. The methods disclosed herein also provide for multiple iterations of binding, observing, and oxidizing for deriving information about multiple targets in a single sample. An associated kit is also provided.