1 July 2009 Concepts for optical high content screens of excitable primary isolated cells for molecular imaging
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Here we describe the cell- and molecular-biological concepts to utilise excitable primary isolated cells, namely cardiomyocytes, for optical high content screens. This starts with an optimised culture of human adult cardiomyocytes, allowing culture with diminished dedifferentiation for one week. To allow fluorescence based molecular imaging genetically encoded biosensors need to be expressed in the cardiomyocytes. For transduction of end-differentiated primary cells such as neurons or cardiomyocytes, a viral gene transfer is necessary. Several viral systems were balanced against each other and an adenoviral system proofed to be efficient. This adenoviral transduction was used to express the calcium sensors YC3.6 and TN-XL in cardiomyocytes. Example measurements of calcium transients were performed by wide-field video imaging. We discuss the potential application of these cellular and molecular tools in basic research, cardiac safety screens and personalised diagnostics.
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Lars Kaestner, Lars Kaestner, Sandra Ruppenthal, Sandra Ruppenthal, Sarah Schwarz, Sarah Schwarz, Anke Scholz, Anke Scholz, Peter Lipp, Peter Lipp, } "Concepts for optical high content screens of excitable primary isolated cells for molecular imaging", Proc. SPIE 7370, Molecular Imaging II, 737008 (1 July 2009); doi: 10.1117/12.831706; https://doi.org/10.1117/12.831706

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