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Author Spotlight: Customized Light-Sheet Imaging for Investigating Myocardial Structures in Rodent Hearts
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Single-Cell Determination of Cardiac Microtissue Structure and Function Using Light Sheet Microscopy.

Diwakar Turaga1, Oriane B Matthys1,2, Tracy A Hookway1

  • 1Gladstone Institutes, San Francisco, California.

Tissue Engineering. Part C, Methods
|March 1, 2020
PubMed
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Light sheet fluorescence microscopy enables single-cell imaging of engineered cardiac microtissues. This advanced technique reveals cell distribution and functional differences, improving our understanding of tissue structure and function.

Area of Science:

  • Biomedical Engineering
  • Cell Biology
  • Microscopy

Background:

  • Native cardiac tissue comprises diverse cell types essential for function.
  • Engineered cardiac tissues require non-myocytes for stability, but their specific roles are unclear.
  • Conventional imaging limits the study of cellular interactions within 3D cardiac microtissues.

Purpose of the Study:

  • To investigate the spatial distribution and functional interactions of multiple cardiac cell types within 3D engineered microtissues.
  • To demonstrate the utility of light sheet fluorescence microscopy (LSFM) for high-resolution imaging of cardiac microtissues.
  • To correlate cellular structure with function at both tissue and single-cell levels.

Main Methods:

  • Utilized light sheet fluorescence microscopy (LSFM) for high-resolution imaging of intact 3D cardiac microtissues.
Keywords:
cardiac microtissuesheterotypic interactionslight sheet microscopylive imagingstructure–function

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  • Performed multicellular spatial distribution analysis of cardiomyocytes (CMs) and cardiac fibroblasts.
  • Conducted live calcium imaging to assess single-cell cardiomyocyte activity within the microtissues.

    • Main Results:

    • CMs and cardiac fibroblasts exhibited random spatial distribution within the 3D microtissues.
    • LSFM enabled single-cell resolution of CM calcium activity.
    • Functional heterogeneity in CM calcium handling correlated with their spatial location in the microtissues.

    Conclusions:

    • LSFM is a powerful tool for analyzing cell-cell interactions and structure-function relationships in engineered cardiac microtissues.
    • Understanding the spatial and functional dynamics of heterogeneous cell populations is crucial for developing advanced cardiac tissue models.
    • This study provides a foundation for future investigations into the complex interplay of cells within engineered tissues.