Fast measurement of sarcomere length and cell orientation in langendorff-perfused hearts using remote focusing microscopy

Edward J. Botcherby, Alex Corbett, Rebecca A.B. Burton, Chris W. Smith, Christian Bollensdorff, Martin J. Booth, Peter Kohl, Tony Wilson, Gil Bub

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    22 Citations (Scopus)

    Abstract

    RATIONALE:: Sarcomere length (SL) is a key indicator of cardiac mechanical function, but current imaging technologies are limited in their ability to unambiguously measure and characterize SL at the cell level in intact, living tissue. OBJECTIVE:: We developed a method for measuring SL and regional cell orientation using remote focusing microscopy, an emerging imaging modality that can capture light from arbitrary oblique planes within a sample. METHODS AND RESULTS:: We present a protocol that unambiguously and quickly determines cell orientation from user-selected areas in a field of view by imaging 2 oblique planes that share a common major axis with the cell. We demonstrate the effectiveness of the technique in establishing single-cell SL in Langendorff-perfused hearts loaded with the membrane dye di-4-ANEPPS. CONCLUSIONS:: Remote focusing microscopy can measure cell orientation in complex 2-photon data sets without capturing full z stacks. The technique allows rapid assessment of SL in healthy and diseased heart experimental preparations.

    Original languageEnglish
    Pages (from-to)863-870
    Number of pages8
    JournalCirculation Research
    Volume113
    Issue number7
    DOIs
    Publication statusPublished - 13 Sep 2013

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    Keywords

    • instrumentation
    • multiphoton fluorescence microscopy
    • optical imaging
    • sarcomeres

    ASJC Scopus subject areas

    • Physiology
    • Cardiology and Cardiovascular Medicine

    Cite this

    Botcherby, E. J., Corbett, A., Burton, R. A. B., Smith, C. W., Bollensdorff, C., Booth, M. J., Kohl, P., Wilson, T., & Bub, G. (2013). Fast measurement of sarcomere length and cell orientation in langendorff-perfused hearts using remote focusing microscopy. Circulation Research, 113(7), 863-870. https://doi.org/10.1161/CIRCRESAHA.113.301704