Measuring the micromechanical properties of embryonic tissues
Measuring the micromechanical properties of embryonic tissues
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DOI：
10.1016/j.ymeth.2015.08.001
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发表时间：
2016-02-01
期刊：
影响因子：
4.8
通讯作者：
Fleury, Vincent
中科院分区：
文献类型：
--
作者：
Chevalier, Nicolas R.;Gazguez, Elodie;Fleury, Vincent
局部力学特性在细胞（分化、迁移）和组织水平（力传递、器官形成、形态发生）上都对胚胎发生起着重要的指导作用。测量它们是一个挑战，因为胚胎组织很小（μ m到mm），柔软（0.1-10千帕）。我们在这里描述了如何制造、校准玻璃纤维悬臂梁，并将其用于施加小力（0.1-10 μ N）、测量收缩活动和评估胚胎组织的总体拉伸弹性。我们概述了压力（流体静力或渗透）如何应用于胚胎组织来量化刚度各向异性。这些技术可以以低成本和最少的设备组装。然后，我们提出了一种方案，准备组织切片，使用原子力显微镜（AFM）进行局部弹性和粘附测量。我们比较了天然和甲醛固定胚胎组织切片的AFM纳米压痕图，并讨论了AFM获得的局部弹性模量与其他体积测量方法获得的弹性模量的比较。我们在小鸡胚胎消化道的具体例子上说明了所有的技术，强调技术问题和常见的陷阱。本报告的主要目的是使生物学家和生物物理学家广泛使用这些微力学测量技术。(C) 2015爱思唯尔公司版权所有。
Local mechanical properties play an important role in directing embryogenesis, both at the cell (differentiation, migration) and tissue level (force transmission, organ formation, morphogenesis). Measuring them is a challenge as embryonic tissues are small (mu m to mm) and soft (0.1-10 kPa). We describe here how glass fiber cantilevers can be fabricated, calibrated and used to apply small forces (0.1-10 mu N), measure contractile activity and assess the bulk tensile elasticity of embryonic tissue. We outline how pressure (hydrostatic or osmotic) can be applied to embryonic tissue to quantify stiffness anisotropy. These techniques can be assembled at low cost and with a minimal amount of equipment. We then present a protocol to prepare tissue sections for local elasticity and adhesion measurements using the atomic force microscope (AFM). We compare AFM nanoindentation maps of native and formaldehyde fixed embryonic tissue sections and discuss how the local elastic modulus obtained by AFM compares to that obtained with other bulk measurement methods. We illustrate all of the techniques presented on the specific example of the chick embryonic digestive tract, emphasizing technical issues and common pitfalls. The main purpose of this report is to make these micromechanical measurement techniques accessible to a wide community of biologists and biophysicists. (C) 2015 Elsevier Inc. All rights reserved.