do Couto Natália Fernanda, Queiroz-Oliveira Thamires, Horta Maria Fátima, Castro-Gomes Thiago, Andrade Luciana Oliveira
Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brasil.
Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brasil.
Bio Protoc. 2020 Aug 5;10(15):e3703. doi: 10.21769/BioProtoc.3703.
Cell signalling, cell secretion, and plasma membrane repair are processes that critically rely on intracellular vesicles, important components of the endocytic and secretory pathways. More specifically, the strategic distribution of intracellular vesicles is important for diverse cellular processes. The method presented here is a simple, affordable, and efficient tool to analyze the distribution of intracellular vesicles such as lysosomes, endosomes, Golgi vesicles or secretory granules under different experimental conditions. The method is an accessible way to analyze the density and dispersion of intracellular vesicles by combining immunofluorescence with pixel-based quantification software (, ImageJ/FIJI). This protocol can be used widely within the scientific community because it utilizes ImageJ/FIJI, an open source software that is free. By tracking fluorescent vesicles based on their position relative to cell nuclei we are able to quantify and analyze their distribution throughout the cell.
细胞信号传导、细胞分泌和质膜修复是严重依赖细胞内囊泡的过程，细胞内囊泡是内吞和分泌途径的重要组成部分。更具体地说，细胞内囊泡的战略分布对多种细胞过程很重要。本文介绍的方法是一种简单、经济且高效的工具，用于分析在不同实验条件下细胞内囊泡（如溶酶体、内体、高尔基体囊泡或分泌颗粒）的分布。该方法是通过将免疫荧光与基于像素的定量软件（如ImageJ/FIJI）相结合来分析细胞内囊泡密度和分散度的一种可行方法。该方案可以在科学界广泛使用，因为它使用的是免费的开源软件ImageJ/FIJI。通过根据荧光囊泡相对于细胞核的位置进行追踪，我们能够量化和分析它们在整个细胞中的分布。
