Transmission electron microscopy (TEM) is a scientific research standard for producing nanometer-resolution ultrastructural images of subcellular components within cells and tissues. Mitochondria, endoplasmic reticulum (ER), lysosomes, and autophagosomes are organelles of particular interest to those investigating metabolic disorders. However, there is no clear consensus amongst regarding the best methods for quantifying the features of organelles in TEM images. In this protocol, we propose a standardized approach to accurately measure the morphology of these important subcellular structures using the free program ImageJ, developed by the National Institutes of Health (NIH). Specifically, we detail procedures for obtaining mitochondrial length, width, area, and circularity, in addition to assessing cristae morphology. We further provide methods for measuring interactions between the mitochondria and ER and measuring the length and width of lysosomes and autophagosomes. This standardized method can be used to quantify key features of organelle morphology, allowing investigators to produce accurate and reproducible measurements of organelle structures in their experimental samples.

中文翻译：

---

使用 ImageJ 分析透射电子显微镜的通用方法

透射电子显微镜 (TEM) 是一种科学研究标准，用于生成细胞和组织内亚细胞成分的纳米分辨率超微结构图像。线粒体、内质网 (ER)、溶酶体和自噬体是那些研究代谢紊乱的人特别感兴趣的细胞器。然而，关于量化 TEM 图像中细胞器特征的最佳方法，目前还没有明确的共识。在该协议中，我们提出了一种标准化方法，使用由美国国立卫生研究院 (NIH) 开发的免费程序 ImageJ 来准确测量这些重要亚细胞结构的形态。具体来说，除了评估嵴形态外，我们还详细介绍了获得线粒体长度、宽度、面积和圆形度的程序。我们进一步提供了测量线粒体和 ER 之间相互作用以及测量溶酶体和自噬体的长度和宽度的方法。这种标准化方法可用于量化细胞器形态的关键特征，使研究人员能够对其实验样品中的细胞器结构进行准确且可重复的测量。