3 years ago

Holographic optical tweezers-based in vivo manipulations in zebrafish embryos

Holographic optical tweezers-based in vivo manipulations in zebrafish embryos
Jana Pfeiffer, Robert Meissner, Sruthi Polali, Erez Raz, Cornelia Denz, Florian Hörner, Timo Betz
Understanding embryonic development requires the characterization of the forces and the mechanical features that shape cells and tissues within the organism. In addition, experimental application of forces on cells and altering cell and organelle shape allows determining the role such forces play in morphogenesis. Here, we present a holographic optical tweezers-based new microscopic platform for in vivo applications in the context of a developing vertebrate embryo that unlike currently used setups allows simultaneous trapping of multiple objects and rapid comparisons of viscoelastic properties in different locations. This non-invasive technique facilitates a dynamic analysis of mechanical properties of cells and tissues without intervening with embryonic development. We demonstrate the application of this platform for manipulating organelle shape and for characterizing the mechanobiological properties of cells in live zebrafish embryos. The method of holographic optical tweezers as described here is of general interest and can be easily transferred to studying a range of developmental processes in zebrafish, thereby establishing a versatile platform for similar investigations in other organisms. Fluorescent beads injected into zebrafish embryos at 1-cell stage are maintained within the embryos and do not affect their development as observed in the presented 1-day old embryo. We established a procedure allowing the use of holographic optical tweezers for the purpose of manipulating cellular organelles and moving particles introduced into cells. The method allows the characterization of mechanical properties distribution within cells, in the physiological environment of the developing early vertebrate embryo. The method is demonstrated by manipulating the shape of the cell nucleus, and by comparing mechanical features at different locations within the cell, using the delay in bead oscillation as readout.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.1002/jbio.201600226

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