Marchi S, Bonora M, Patergnani S, Giorgi C, Pinton P
Section of Pathology, Oncology and Experimental Biology, Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, Ferrara, Italy.
Section of Pathology, Oncology and Experimental Biology, Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, Ferrara, Italy.
Methods Enzymol. 2017;588:171-186. doi: 10.1016/bs.mie.2016.09.080. Epub 2016 Nov 12.
It is widely acknowledged that mitochondria are highly active structures that rapidly respond to cellular and environmental perturbations by changing their shape, number, and distribution. Mitochondrial remodeling is a key component of diverse biological processes, ranging from cell cycle progression to autophagy. In this chapter, we describe different methodologies for the morphological study of the mitochondrial network. Instructions are given for the preparation of samples for fluorescent microscopy, based on genetically encoded strategies or the employment of synthetic fluorescent dyes. We also propose detailed protocols to analyze mitochondrial morphometric parameters from both three-dimensional and bidimensional datasets. Finally, we describe a protocol for the visualization and quantification of mitochondrial structures through electron microscopy.
人们普遍认为，线粒体是高度活跃的结构，可通过改变其形状、数量和分布来快速响应细胞和环境扰动。线粒体重塑是从细胞周期进程到自噬等多种生物学过程的关键组成部分。在本章中，我们描述了用于线粒体网络形态学研究的不同方法。基于基因编码策略或使用合成荧光染料，给出了荧光显微镜样品制备的说明。我们还提出了详细的方案，用于从三维和二维数据集中分析线粒体形态计量参数。最后，我们描述了一种通过电子显微镜可视化和定量线粒体结构的方案。
