Melo Pedro, S Silveira Mariana, Mendes-Pinto Inês, Relvas João B
Instituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, Porto, Portugal.
Instituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, Porto, Portugal; Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Methods Cell Biol. 2023;174:75-92. doi: 10.1016/bs.mcb.2022.08.003. Epub 2022 Sep 10.
Microglia cells dynamically survey the central nervous system microenvironment and, in response to tissue damage inflicted by radiation therapy, disease or infection, undergo morphological and functional changes that culminate in microglia activation. Cell shape transformation can be assessed descriptively or, alternatively, it can be quantified as a continuous variable for parameters including total cell size as well as protrusion length, ramification and complexity. The purpose of the MorphoMacro method is to quantitatively profile multiple and single microglia cells using the available ImageJ platform. This method outlines the required steps and ImageJ plugins to convert fluorescence and bright-field photomicrographs into representative binary and skeletonized images and to analyze them using the MorphoMacro software plugin for multiparametric and multilevel description of microglia cell morphology in vivo and ex vivo. Overall, the protocol provides a quantitative and comprehensive tool that can be used to identify, stratify, and monitor diverse microglia morphologies in homeostatic, different disease conditions and subsequent therapeutic monitoring.
小胶质细胞动态监测中枢神经系统微环境，并在受到放射治疗、疾病或感染造成的组织损伤时，经历形态和功能变化，最终导致小胶质细胞激活。细胞形状转变可以进行描述性评估，或者也可以将其量化为一个连续变量，用于包括细胞总体大小以及突起长度、分支和复杂性等参数。MorphoMacro方法的目的是利用现有的ImageJ平台对多个和单个小胶质细胞进行定量分析。该方法概述了将荧光和明场显微照片转换为代表性的二值化和骨架化图像，并使用MorphoMacro软件插件对体内和体外小胶质细胞形态进行多参数和多层次描述所需的步骤和ImageJ插件。总体而言，该方案提供了一种定量且全面的工具，可用于在稳态、不同疾病状态及后续治疗监测中识别、分层和监测多种小胶质细胞形态。
