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SAN DIEGO, CA, United States

Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase II | Award Amount: 761.59K | Year: 2016

DESCRIPTION provided by applicant Defects in heart formation during embryonic and fetal development likely contribute to miscarriages which terminate up to of all pregnancies and Congential Heart Defects CHDs which are among the most common birth defects Environmental factors may influence miscarriages and up to of all CHDs but little is known about how such chemicals affect formation of the heart during early embryonic differentiation The goal of this SBIR project is to develop a Cardiopoiesis Assay heart formation assay to enable testing of compounds to determine if they influence the emergence of cardiac myocytes from multipotent mesodermal progenitors a critical early decision point in heart formation During the previous Phase I portion of the project a version of the assay was developed using mouse embryonic stem cells and this assay has proven useful in identifying chemicals siRNAs and miRs that modify cardiac differentiation For Phase II we propose to develop the assay using human induced pluripotent stem cells hiPSCs which will be derived from multiple donors to increase the genetic diversity represented by the assay The Cardiopoiesis Assay will be useful in predicting whether chemical compounds affect heart formation in the human embryo which is of critical importance to human health The assay will be of interest to government agencies in the US such as the Environmental Protection Agency and worldwide concerned with environmental toxicology PUBLIC HEALTH RELEVANCE Defects of the heart are the most common birth defects and may be caused in part by toxic compounds from the environment that interfere with heart formation prior to birth We propose to develop a test system to screen chemicals for dangerous effects on heart formation We propose to use human stem cells which we will derive from skin samples obtained from volunteer donors the skin samples are obtained by a painless procedure that is risk free Such stem cells can be cultured in a manner that they form beating heart cells in laboratory dishes We will develop a procedure to test if chemicals prevent formation of heart cells from stem cells as such chemicals are likely to cause heart birth defects

Automated image screening operations of pipelined image processing systems and methods are controlled with a graphical user interface enabling user control of screening analysis process setup and execution and user viewing of results. A gating interface is provided to include and/or exclude cells in an aggregation of individual cell data.

Video recordings from two or more optical channels are produced, processed, and analyzed simultaneously in order to provide quantitative analysis of action potentials, calcium transients and ionic flux in excitable cells loaded with voltage or ion sensitive dyes with distinct excitation and emission wavelengths, The specific wavelengths of fluorescent light emitted from each dye are separated and recorded. The recordings are mutually registered and cytometric analysis is performed to provide a quantitative analysis of the action potentials calcium transient, and/or ionic flux on a cell-by-cell and well-by-well basis in microtiter plates. The cells are then fixed, labeled for other biomarkers, and scanned again. The resulting fixed cell images are registered with the live cell recordings and analyzed; missing cells that were washed off are detected relative to the live recordings, and cytometry data from live and fixed cell scans is collated cell-by-cell.

A system, method and kit for processing an original image of biological material to identify certain components of a biological object by locating the biological object in the image, enhancing the image by sharpening components of interest in the object, and applying a contour-finding function to the enhanced image to create a contour mask. The contour mask may be processed to yield a segmented image divided by structural units of the biological material.

Continuous-scanning image acquisition in an automated microscopy system uses an image reflected off of an object that supports a specimen being imaged to automatically focus the microscopy system.

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