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Updated: Mar 13, 2026

Imaging Cleared Embryonic and Postnatal Hearts at Single-cell Resolution
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Imaging Cleared Embryonic and Postnatal Hearts at Single-cell Resolution.

Wasay M Shaikh Qureshi1, Lianjie Miao1, David Shieh1

  • 1Department of Molecular and Cellular Physiology, Albany Medical College.

Journal of Visualized Experiments : Jove
|October 22, 2016
PubMed
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Researchers developed new whole-embryo clearing methods to visualize single cardiac progenitor cells. This technique enables detailed study of heart development and congenital heart defects at single-cell resolution.

Area of Science:

  • Developmental Biology
  • Cell Biology
  • Biotechnology

Background:

  • Whole-heart clonal analysis is crucial for understanding cardiac development and morphogenesis.
  • Existing imaging techniques are limited by tissue opacity and light scattering at increased depths.
  • New clearing methods are needed to achieve single-cell resolution in whole-heart imaging.

Purpose of the Study:

  • To develop and adapt whole-embryo clearing techniques for high-resolution imaging of cardiac morphogenesis.
  • To evaluate the efficacy of Scale and CUBIC clearing methods for embryonic and postnatal hearts.
  • To enable the study of gene function at a single-clone resolution during cardiac development.

Main Methods:

  • Established single-cell lineage tracing using the ROSA26-CreERT2; ROSA26-Confetti system.

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Last Updated: Mar 13, 2026

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  • Adapted and combined whole-embryo clearing methodologies (Scale, CUBIC) with whole-mount staining.
  • Imaged single clones within the heart at single-cell resolution.

    • Main Results:

    • Successfully imaged the heart at single-cell resolution using adapted clearing techniques.
    • Scale effectively cleared embryonic hearts but not postnatal hearts.
    • CUBIC cleared postnatal hearts but caused damage to embryonic hearts.

    Conclusions:

    • The described methods allow for the study of cardiac morphogenesis at single-cell resolution.
    • Optimized clearing techniques are essential for overcoming imaging limitations in whole-heart studies.
    • This approach will advance the understanding of the cellular and molecular basis of congenital heart defects.