42 results about "High-content screening" patented technology

The present invention provides protein fragment complementation assays for drug discovery, in particular to identify compounds that activate or inhibit cellular pathways. Based on the selection of an interacting protein pair combined with an appropriate PCA reporter, the assays may be run in high-throughput or high-content mode and may be used in automated screening of libraries of compounds. The interacting pair may be selected by cDNA library screening; by gene-by-gene interaction mapping; or by prior knowledge of a pathway. Fluorescent and luminescent assays can be constructed using the methods provided herein. The selection of suitable PCA reporters for high-throughput or high-content (high-context) assay formats is described for a diversity of reporters, with particular detail provided for examples of monomeric enzymes and fluorescent proteins. Methods are described for constructing such assays for one or more steps in a biochemical pathway; testing the effects of compounds from combinatorial, natural product, peptide, antibody, nucleic acid or other diverse libraries on the protein or pathway(s) of interest; and using the results of the screening to identify specific compounds that activate or inhibit the protein or pathway(s) of interest. Single-color and multi-color assays are disclosed. Further disclosed are universal expression vectors with cassettes that allow the rapid construction of assays for a large and diverse number of gene / reporter combinations. The development of such assays is shown to be straightforward, providing for a broad, flexible and biologically relevant platform for drug discovery.

The present invention relates generally to the field of high content screening of particles, e.g., cells in a flow cytometric system. In particular, the present invention relates to devices, methods and systems to obtain line-scan images of particles, e.g., cells, of a plurality of different samples simultaneously, where the line-scan images can be used to identify cells based on at least one of a variety of phenotypic characteristics such as shape, asymmetry, and intracellular information for cell sorting and selection. In some embodiments, the line-scan images are obtained as the particles, e.g., cells, in a plurality of different samples flow through a plurality of microchannels, reducing the need and time for focusing of the image detection system. In some embodiments, the laser spot size has a small spatial resolution for rapid capturing of images of cells. In some embodiments, the laser spot size has a larger spatial resolution for imaging of larger particles or cells, e.g., rare cells in a sample. In some embodiments, the devices, methods and systems are automated for a high-throughput, high content screening of particles such as cells in a plurality of samples.

Disclosed are intra-culture perfusion methods applied to standard cell culture disposables using three-dimensional cell culture scaffold and synthetic vasculature compositions in high-throughput screening and high-content screening applications, and assay development.

Systems and methods are provided for characterizing a multidimensional distribution of responses from the objects in a populations subject to a perturbation. The methods enable the creation of a “degree of response” scale interpolated from non-perturbed and perturbed reference populations. The methods enables, using the interpolated degree of response scale, the quantitation of a degree of response of a test compound subject to a given level of perturbation, and enables the generation of a dose-response curve for a test compound. The methods are useful in a wide range of applications, such cellular analysis and high-content screening of compounds, as carried out in pharmaceutical research.

Microplates containing spherical “microsphere” fluorescence standards are disclosed. The microplates can be prepared using several methods including airbrushing, application with an inkjet printer, or controlled evaporation. Spherical bead standards containing two or more regions stained with dyes of different fluorescence lifetimes, and methods for their preparation are also disclosed. The microplates can be used as calibration standards for fluorescence and confocal microscopes, and as calibration tools for microscope-based high content screening (“HCS”) instruments.

Data processing system for generating representations and analyses of cytometric information for:—loading one set of data including:—one image of two wells on a plate each having one cell in a predetermined biological condition, each image being acquired with a microscope;—features of each cell from each image;—processing loaded data for obtaining representations and analyses of the data, from:—Histogram in one dimension;—Scattergram in two-dimensions;—Density in two-dimensions;—Plate layout management;—Cell montage;—Field;—Plate heatmap;—Dose response with regrouping of replicates;—Statistics information percentage of cell in different gates;—zFactor calculation with regrouping of replicates;—List of well for selection and list of biological condition for selection;—Statistics export capability;—Ready to print layout capability on the selected well or biological condition or all wells and biological conditions.

The invention relates to an establishing method and application of a dilated cardiomyopathy and zebrafish disease model. A disease model simulating human congenital dilated cardiomyopathy (CDCM) is firstly established; and the disease model can be used as a drug screening model to be applied to screening of drugs capable of relieving symptoms of heart failure caused by CDCM. The method provided by the invention is an economic and rapid high-content screening method; and the obtained compound has relatively high specificity and druggability.

Disclosed herein are systems methods for high content screening for microscope imaging. In some embodiments, the system comprises: a microscope; and a processor configured to implement: a slide loader module; a reference imager module; a slide imager module; a region of interest (ROI) finder module; a compare imager module; a calibrator module; and an image stitcher module.

A method and system for setting image analysis parameters to control image analysis operations. The method and system include collecting set of digital training images including a set of states for the set of digital training images. An objective function is defined to determine a relative quality of plural different parameter sets used for digital image analysis. Values for the plural different parameter sets that maximize (or minimize) the objective function are determined. The method and system increases a usability of high content screening technologies by reducing a required level of expertise required to configure digital image processing.

The present invention involves the pooling of multiple high content cell-based screening assays, and carrying out a primary screen in a one or more channels of a fluorescence detection device, which drastically increases the number of simultaneous high content cell-based screening events that can be carried out. Subsequent deconvolution of primary screen “hits” (ie: those wells or locations on an array of locations in which the one or more test compounds caused a change in the fluorescence signal(s) from the fluorescent reporter molecules in the cells) enables much more rapid generation of high content cell screening data than was previously possible, and at significantly reduced costs.

The present invention belongs to the field of pharmacy and cell biology, and relates to an efficient and reliable high content CCR4 antagonist screening cell model using human chemokine receptor 4 as a target, and a screening method thereof. According to the present invention, a cell line being subjected to human CCR4-enhanced green fluorescent protein fusion expression introducing is established, excitation is performed through human thymus activation regulated chemokine and human macrophage chemotatic factor, the CCR4 with green fluorescent protein is induced to produce re-distribution so as to form green fluorescent particles, and the CCR4 antagonist screening cell model for high-content screening is established; the bioactivity of the compound is evaluated by determining the CCR4 green fluorescent particle formation excited or inhibited by the screened compound through the model; and the established drug screening model is the sensitive, efficient and reliable CCR4 antagonist screening cell model suitable for high-throughput and high-content screening, and the method is the sensitive, efficient and reliable method suitable for high-throughput and high-content screening.

The invention discloses a cell phenotype based high-content multi-index renal toxicity detection method and applications thereof. According to the detection method, A fluorescent molecular probe that can change the functions of nucleus or mitochondrial and carry out specific detection is adopted to label cells; a high content screening system is taken as the detection means, based on the high content screening system, live cells, which are injected with drugs to be detected, are subjected to imaging analysis so as to determine at least one influence of the drugs on cell number, nucleus and mitochondrial, and thus the influence of the drugs on the cells can be determined. The invention also discloses applications of the detection method. The provided detection method has the advantages that multiple indexes can be detected, while the cell structure and functional integrity are maintained, and the detection method has the characteristics of rapidness, high flux, low cost, and high accuracy.

The present invention is directed to Protein-fragment Complementation Assays (PCAs) and assay compositions based on fluorescent proteins. The invention provides methods for fragmenting fluorescent proteins and generating mutant fragments with desired spectral characteristics for PCA. The invention encompasses assays and compositions based on fluorescent proteins from the species Aequorea, Anemonia and Anthozoa. In particular, the invention is directed to fragments of mutant fluorescent proteins having improved spectral properties over the wild-type proteins. The invention encompasses fragments of mutant versions of A. Victoria green fluorescent protein (GFP), in particular yellow fluorescent proteins (EYFP and super-EYFP), ‘Venus’, cyan, ‘citrine’, blue, cyan-green, and photoactivatable variants of GFP The invention also encompasses red fluorescent PCAs based on Discosoma red fluorescent protein (RFP PCA)and a kindling fluorescent protein PCA (KFP1 PCA) derived from Anemonia sulcata. Any useful mutation of a fluorescent protein can be engineered into a fragment, generating a wide range of assays useful for drug discovery, target validation, high-throughput screening, high-content screening, pathway mapping, drug mechanism-of-action studies, biosensors, and diagnostics.

A method and system for setting image analysis parameters to control image analysis operations. The method and system include collecting set of digital training images including a set of states for the set of digital training images. An objective function is defined to determine a relative quality of plural different parameter sets used for digital image analysis. Values for the plural different parameter sets that maximize (or minimize) the objective function are determined. The method and system increases a usability of high content screening technologies by reducing a required level of expertise required to configure digital image processing.

In one aspect, the present invention relates to a method 200 for identifying one or more phenotypes from a multi-parameter data set. The method 200 comprises measuring 202 correlation between pairs of parameters within the multi-parameter data set, modifying 204 correlated parameter values within a predetermined multi-parameter data analysis set to form an analysis parameter set, and analyzing 206 the multi-parameter data set using the analysis parameter set to identify one or more phenotypes from the multi-parameter data set. Various embodiments of the present invention may, for example, be used in an automated high-content screening (HCS) apparatus 100 for biological cellular analysis.

Described herein is a three-dimensional cell culture scaffold composition comprising an absorbent rigid (AR) component, and in some embodiments, further comprises a gel component. The absorbent rigid component preferably comprises a glass fiber material. It is a surprising finding of the present invention that an AR component having a void volume of between approximately 70% and 95% results in a three-dimensional cell culture composition that allows for robust, high-throughput screening and high-content screening accessible tissue models with preserved cell morphology, heterogeneity of cell types and cell populations, extracellular matrix constituents, functional cell-cell and cell-extracellular matrix interactions and signaling with sufficient specificities to tissue physiology and pathology.

The invention relates to an establishing method and application of a dilated cardiomyopathy and zebrafish disease model. A disease model simulating human congenital dilated cardiomyopathy (CDCM) is firstly established; and the disease model can be used as a drug screening model to be applied to screening of drugs capable of relieving symptoms of heart failure caused by CDCM. The method provided by the invention is an economic and rapid high-content screening method; and the obtained compound has relatively high specificity and druggability.

Disclosed are intra-culture perfusion methods applied to standard cell culture disposables using three-dimensional cell culture scaffold and synthetic vasculature compositions in high-throughput screening and high-content screening applications, and assay development.