Imaging Facility, Max Planck Institute of Immunobiology and Epigenetics, Freiburg, Germany.
PLoS One. 2018 Sep 18;13(9):e0203956. doi: 10.1371/journal.pone.0203956. eCollection 2018.
Mitochondrial DNA molecules coated with proteins form compact particles called mitochondrial nucleoids. They are redistributed within mitochondrial network undergoing morphological changes. The straightforward technique to characterize nucleoids' motions is fluorescence microscopy. Mitochondrial nucleoids are commonly labelled with fluorescent protein tags, which is not always feasible and was reported to cause artifacts. Organic DNA-binding dyes are free of these drawbacks, but they lack specificity to mitochondrial DNA. Here, considering physico-chemical properties of such dyes, we achieved preferential live-cell labelling of mitochondrial nucleoids by a nucleic acid staining dye SYBR Gold. It enabled time-lapse imaging of mitochondrial nucleoids by structured illumination microscopy and quantification of their motions.
线粒体 DNA 分子与蛋白质结合形成称为线粒体类核的致密颗粒。它们在经历形态变化的线粒体网络中重新分布。描述类核运动的直接技术是荧光显微镜。线粒体类核通常用荧光蛋白标签标记,但并非总是可行,并且据报道会造成假象。有机 DNA 结合染料没有这些缺点,但它们对线粒体 DNA 没有特异性。在这里,考虑到这些染料的物理化学性质,我们通过核酸染色染料 SYBR Gold 实现了线粒体类核的优先活细胞标记。这使得通过结构光照明显微镜对线粒体类核进行延时成像,并对它们的运动进行定量。