Faculty of Health and Life Sciences, Biosciences, University of Exeter, Exeter, Devon, UK.
Methods Mol Biol. 2023;2643:105-122. doi: 10.1007/978-1-0716-3048-8_8.
Transmission electron microscopy (TEM) has long been a vital technology to visualize the interaction of cellular compartments at the highest possible resolution. While this paved the way to describing organelles within the cellular context in detail, TEM has long been underused to generate quantitative data, analyzing those interactions as well as underlying mechanisms leading to their formation and modification. Here we describe a simple stereological method to unbiasedly assess the extent of organelle-organelle membrane contact sites, able to efficiently generate accurate and reproducible quantitative data from cultured mammalian cells prepared for TEM.
透射电子显微镜（TEM）长期以来一直是一种至关重要的技术，可以以最高可能的分辨率可视化细胞区室的相互作用。虽然这为详细描述细胞环境中的细胞器铺平了道路，但 TEM 长期以来一直没有被充分用于生成定量数据，也没有分析这些相互作用以及导致它们形成和修饰的潜在机制。在这里，我们描述了一种简单的体视学法，以无偏地评估细胞器-细胞器膜接触位点的程度，这种方法能够从为 TEM 准备的培养的哺乳动物细胞中高效地生成准确和可重复的定量数据。
