DNA Damage and Repair

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Poster:
Multispectral imaging flow cytometry reveals distinct frequencies of γ-H2AX foci induction in DNA double strand break repair defective human cell lines.— Bourton EC. et.al. Cytometry A. 2012 Feb;81(2):130-7. doi: 10.1002
  Technical Information:
Quantitation of gamma-H2AX spots on the ImageStream (09-008)
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Merck:/Freestyle/BI-Bioscience/Cell-Analysis/amnis/Amins2-images/Application-Covers/2014-Rodrigues-CytoA-1.jpgReference Paper:
Multi-Parameter Dose Estimations in Radiation Biodosimetry Using the Automated Cytokinesis-Block Micronucleus Assay with Imaging Flow Cytometry. Cytometry A 2014; 10: 883-893.


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Merck:/Freestyle/BI-Bioscience/Cell-Analysis/amnis/Amins2-images/dna-damage-2.jpgThe study of DNA repair mechanisms is important in the field of oncology where radiotherapy used to treat tumors induces double strand breaks (DSB). The DNA damage that occurs can be indirectly visualized with a microscope by immunostaining the repair proteins that are recruited to DSB foci. Amnis® imaging flow cytometry automatically and objectively collect thousands of images, quantifies the foci in a population of cells and determines dose response kinetics faster and easier than manual microscopy.


Quantifying Micronuclei in Irradiated Human Lymphocytes Using the ImageStream®X

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The cytokinesis-block micronucleus (CBMN) assay is a well-established technique in biological dosimetry for estimating radiation doses by correlating the frequency of MN in binucleated cells (BNCs) to a calibrated dose in peripheral blood lymphocytes. Advanced masking techniques allow for automated imaging, identification and enumeration of fluorescently stained BNCs and MN in irradiated lymphocytes. Using features such as aspect ratio, shape ratio and fluorescence spot counting, rapid analysis can be performed on large numbers of cell images captured by the ImageStream®X. Data was generously provided by M. A. Rodrigues.

Quantitation of γ-H2AX foci in Irradiated Cells Using the ImageStream®X

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Phosphorylated H2AX (γ-H2AX) facilitates recognition and repair of DNA double strand breaks (DSBs) that may occur from exposure to ionizing radiation. Staining irradiated cells for γ-H2AX reveals nuclear foci that are readily observed microscopically in a dose response manner. Irradiated cells were analyzed for the number of spots in the nuclear region using advanced masking techniques that identify the punctate staining. Morphological measurements employed in this analysis including object shape, size, and punctate fluorescence spot counting emphasizing the advantages of quantitative multiparametric image analysis on large numbers of cells provided with the ImageStream®X.