Dynamic structure and protein expression of the live embryonic heart captured by 2-photon light sheet microscopy and retrospective registration

Creators: Trivedi, Vikas; Truong, Thai V.; Trinh, Le A.; Holland, Daniel B.; Liebling, Michael; Fraser, Scott E.

Abstract

We present an imaging and image reconstruction pipeline that captures the dynamic three-dimensional beating motion of the live embryonic zebrafish heart at subcellular resolution. Live, intact zebrafish embryos were imaged using 2-photon light sheet microscopy, which offers deep and fast imaging at 70 frames per second, and the individual optical sections were assembled into a full 4D reconstruction of the beating heart using an optimized retrospective image registration algorithm. This imaging and reconstruction platform permitted us to visualize protein expression patterns at endogenous concentrations in zebrafish gene trap lines.

Additional Information

© 2015 Optical Society of America. Received 12 Mar 2015; revised 25 Apr 2015; accepted 1 May 2015; published 11 May 2015. This work was funded by Rosen Center for Bioengineering, Caltech, Center of Excellence in Genomic Science (Caltech), NIH/NHGRI Grant # P50HR004071 and FaceBase, NIH Grant # U01DE020063. We thank Leigh Ann Fletcher, Colleen Paquette and Andrey Andreev for fish care.

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