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

    Research output: Contribution to journalArticle

    19 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

    Fingerprint

    Sarcomeres
    Microscopy
    Photons
    Isolated Heart Preparation
    Heart Diseases
    Coloring Agents
    Technology
    Light
    Membranes

    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., ... 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

    Fast measurement of sarcomere length and cell orientation in langendorff-perfused hearts using remote focusing microscopy. / Botcherby, Edward J.; Corbett, Alex; Burton, Rebecca A B; Smith, Chris W.; Bollensdorff, Christian; Booth, Martin J.; Kohl, Peter; Wilson, Tony; Bub, Gil.

    In: Circulation Research, Vol. 113, No. 7, 13.09.2013, p. 863-870.

    Research output: Contribution to journalArticle

    Botcherby, EJ, Corbett, A, Burton, RAB, Smith, CW, Bollensdorff, C, Booth, MJ, 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, vol. 113, no. 7, pp. 863-870. https://doi.org/10.1161/CIRCRESAHA.113.301704
    Botcherby, Edward J. ; Corbett, Alex ; Burton, Rebecca A B ; Smith, Chris W. ; Bollensdorff, Christian ; Booth, Martin J. ; Kohl, Peter ; Wilson, Tony ; Bub, Gil. / Fast measurement of sarcomere length and cell orientation in langendorff-perfused hearts using remote focusing microscopy. In: Circulation Research. 2013 ; Vol. 113, No. 7. pp. 863-870.
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    AU - Bollensdorff, Christian

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