Marsal Maria, Jorba Ignasi, Rebollo Elena, Luque Tomas, Navajas Daniel, Martín-Blanco Enrique
Instituto de Biología Molecular de Barcelona, Consejo Superior de Investigaciones Científicas.
Institute for Bioengineering of Catalonia, Universitat de Barcelona and CIBER Enfermedades Respiratorias.
J Vis Exp. 2017 Nov 29(129):56224. doi: 10.3791/56224.
Elucidating the factors that direct the spatio-temporal organization of evolving tissues is one of the primary purposes in the study of development. Various propositions claim to have been important contributions to the understanding of the mechanical properties of cells and tissues in their spatiotemporal organization in different developmental and morphogenetic processes. However, due to the lack of reliable and accessible tools to measure material properties and tensional parameters in vivo, validating these hypotheses has been difficult. Here we present methods employing atomic force microscopy (AFM) and particle tracking with the aim of quantifying the mechanical properties of the intact zebrafish embryo yolk cell during epiboly. Epiboly is an early conserved developmental process whose study is facilitated by the transparency of the embryo. These methods are simple to implement, reliable, and widely applicable since they overcome intrusive interventions that could affect tissue mechanics. A simple strategy was applied for the mounting of specimens, AFM recording, and nanoparticle injections and tracking. This approach makes these methods easily adaptable to other developmental times or organisms.
阐明指导不断演变的组织的时空组织的因素是发育研究的主要目的之一。各种观点声称对理解细胞和组织在不同发育和形态发生过程中的时空组织的力学特性做出了重要贡献。然而，由于缺乏可靠且易于使用的工具来测量体内的材料特性和张力参数，验证这些假设一直很困难。在此，我们介绍了采用原子力显微镜（AFM）和粒子追踪的方法，旨在量化斑马鱼胚胎卵黄细胞在胚盘形成期间的完整力学特性。胚盘形成是一个早期保守的发育过程，胚胎的透明性有助于对其进行研究。这些方法易于实施、可靠且广泛适用，因为它们克服了可能影响组织力学的侵入性干预。我们应用了一种简单的策略来进行样本固定、AFM记录以及纳米颗粒注射和追踪。这种方法使这些方法能够轻松适应其他发育阶段或生物体。
