Methods for dynamic and whole volume imaging of the zebrafish heart

Isaac Bakis; Yuhan Sun; Laila Abd Elmagid; Xidi Feng; Mher Garibyan; Joycelyn K Yip; Fang Zhou Yu; Sayali Chowdhary; Gerardo Esteban Fernandez; Jingli Cao; Megan L McCain; Ching-Ling Lien; Michael Rm Harrison

doi:10.1016/j.ydbio.2023.09.002

Methods for dynamic and whole volume imaging of the zebrafish heart

Dev Biol. 2023 Dec:504:75-85. doi: 10.1016/j.ydbio.2023.09.002. Epub 2023 Sep 12.

Authors

Affiliations

¹ Cardiovascular Research Institute, Weill Cornell Medicine, New York, NY, 10021, USA; Department of Cell and Developmental Biology, Weill Cornell Medicine, 1300 York Avenue, New York, NY, 10021, USA.
² Saban Research Institute and Heart Institute of Children's Hospital Los Angeles, Los Angeles, CA, 90027, USA.
³ Laboratory for Living Systems Engineering, Alfred E. Mann Department of Biomedical Engineering, USC Viterbi School of Engineering, University of Southern California, Los Angeles, CA, 90089, USA.
⁴ Cardiovascular Research Institute, Weill Cornell Medicine, New York, NY, 10021, USA; Department of Emergency Medicine, Nuvance Health, Poughkeepsie, NY, 12601, USA.
⁵ Laboratory for Living Systems Engineering, Alfred E. Mann Department of Biomedical Engineering, USC Viterbi School of Engineering, University of Southern California, Los Angeles, CA, 90089, USA; Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA, 90033, USA.
⁶ Saban Research Institute and Heart Institute of Children's Hospital Los Angeles, Los Angeles, CA, 90027, USA; Department of Surgery, Keck School of Medicine, University of Southern California, Los Angeles, CA, 90033, USA. Electronic address: clien@chla.usc.edu.
⁷ Cardiovascular Research Institute, Weill Cornell Medicine, New York, NY, 10021, USA; Department of Cell and Developmental Biology, Weill Cornell Medicine, 1300 York Avenue, New York, NY, 10021, USA. Electronic address: mrh4003@med.cornell.edu.

PMID: 37708968
PMCID: PMC10841891 (available on 2024-12-01)
DOI: 10.1016/j.ydbio.2023.09.002

Abstract

Tissue development and regeneration are dynamic processes involving complex cell migration and cell-cell interactions. We have developed a protocol for complementary time-lapse and three-dimensional (3D) imaging of tissue for developmental and regeneration studies which we apply here to the zebrafish cardiac vasculature. 3D imaging of fixed specimens is used to first define the subject at high resolution then live imaging captures how it changes dynamically. Hearts from adult and juvenile zebrafish are extracted and cleaned in preparation for the different imaging modalities. For whole-mount 3D confocal imaging, single or multiple hearts with native fluorescence or immuno-labeling are prepared for stabilization or clearing, and then imaged. For live imaging, hearts are placed in a prefabricated fluidic device and set on a temperature-controlled microscope for culture and imaging over several days. This protocol allows complete visualization of morphogenic processes in a 3D context and provides the ability to follow cell behaviors to complement in vivo and fixed tissue studies. This culture and imaging protocol can be applied to different cell and tissue types. Here, we have used it to observe zebrafish coronary vasculature and the migration of coronary endothelial cells during heart regeneration.

Keywords: Antibody labeling; Clearing; Coronary vessels; Imaging; Live-cell imaging; Regeneration; Zebrafish.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Endothelial Cells* / metabolism
Heart / diagnostic imaging
Imaging, Three-Dimensional / methods
Zebrafish*

Abstract

Publication types

MeSH terms

Grants and funding