Hörner Florian, Meissner Robert, Polali Sruthi, Pfeiffer Jana, Betz Timo, Denz Cornelia, Raz Erez
Institute of Cell Biology, Center of Molecular Biology of Inflammation, University of Münster, Von-Esmarch-Straße 56, 48149, Münster, Germany.
Institute of Applied Physics, University of Münster, Corrensstraße 2/4, 48149, Münster, Germany.
J Biophotonics. 2017 Nov;10(11):1492-1501. doi: 10.1002/jbio.201600226. Epub 2017 Feb 6.
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.
理解胚胎发育需要对塑造生物体内细胞和组织的力及机械特征进行表征。此外，对细胞施加力并改变细胞和细胞器形状的实验应用能够确定这些力在形态发生中所起的作用。在此，我们展示了一种基于全息光镊的新型显微平台，用于发育中的脊椎动物胚胎的体内应用，与目前使用的装置不同，该平台允许同时捕获多个物体，并能快速比较不同位置的粘弹性特性。这种非侵入性技术有助于在不干扰胚胎发育的情况下对细胞和组织的力学特性进行动态分析。我们展示了该平台在操纵细胞器形状以及表征活斑马鱼胚胎中细胞的力学生物学特性方面的应用。这里所描述的全息光镊方法具有普遍意义，并且可以很容易地转移到研究斑马鱼的一系列发育过程中，从而为其他生物体的类似研究建立一个通用平台。如在呈现的1天大的胚胎中所观察到的，在单细胞阶段注射到斑马鱼胚胎中的荧光珠会保留在胚胎内且不影响其发育。
