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Light-sheet fluorescence imaging to localize cardiac lineage and protein distribution.

  • Author(s): Ding, Yichen
  • Lee, Juhyun
  • Ma, Jianguo
  • Sung, Kevin
  • Yokota, Tomohiro
  • Singh, Neha
  • Dooraghi, Mojdeh
  • Abiri, Parinaz
  • Wang, Yibin
  • Kulkarni, Rajan P
  • Nakano, Atsushi
  • Nguyen, Thao P
  • Fei, Peng
  • Hsiai, Tzung K
  • et al.

Published Web Location

https://doi.org/10.1038/srep42209
Abstract

Light-sheet fluorescence microscopy (LSFM) serves to advance developmental research and regenerative medicine. Coupled with the paralleled advances in fluorescence-friendly tissue clearing technique, our cardiac LSFM enables dual-sided illumination to rapidly uncover the architecture of murine hearts over 10 by 10 by 10 mm3 in volume; thereby allowing for localizing progenitor differentiation to the cardiomyocyte lineage and AAV9-mediated expression of exogenous transmembrane potassium channels with high contrast and resolution. Without the steps of stitching image columns, pivoting the light-sheet and sectioning the heart mechanically, we establish a holistic strategy for 3-dimentional reconstruction of the "digital murine heart" to assess aberrant cardiac structures as well as the spatial distribution of the cardiac lineages in neonates and ion-channels in adults.

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