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Published August 2008 | Published
Journal Article Open

An all-optical approach for probing microscopic flows in living embryos

Abstract

Living systems rely on fluid dynamics from embryonic development to adulthood. To visualize biological fluid flow, devising the proper labeling method compatible with both normal biology and in vivo imaging remains a major experimental challenge. Here, we describe a simple strategy for probing microscopic fluid flows in vivo that meets this challenge. An all-optical procedure combining femtosecond laser ablation, fast confocal microscopy and 3D-particle tracking was devised to label, image and quantify the flow. This approach is illustrated by studying the flow generated within a micrometer scale ciliated vesicle located deep inside the zebrafish embryo and involved in breaking left-right embryonic symmetry. By mapping the velocity field within the vesicle and surrounding a single beating cilium, we show this method can address the dynamics of cilia-driven flows at multiple length scales, and can validate the flow features as predicted from previous simulations. This approach provides new experimental access to questions of microscopic fluid dynamics in vivo.

Additional Information

© 2008 Biophysical Society. Received for publication 16 May 2008 and in final form 6 June 2008. We are grateful to D. Wu and T. Truong for critical comments, the Caltech Biological Imaging Center, and P. Björkman for sharing equipments. J.V. was supported by the Human Frontier Science Program Fellowship. This work was supported by the Biological Imaging Center of the Caltech Beckman Institute and a Centers of Excellence in Genomic Sciences grant from the National Institutes of Health (P50 HG004071).

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