Le Gall Antoine, Cattoni Diego I, Nollmann Marcelo
Centre de Biochimie Structurale, CNRS UMR5048, INSERM U1054, Université de Montpellier, 29 rue de Navacelles, 34090, Montpellier, France.
Methods Mol Biol. 2017;1624:253-268. doi: 10.1007/978-1-4939-7098-8_19.
The bacterial nucleoid is highly organized, yet it is dynamically remodeled by cellular processes such as transcription, replication, or segregation. Many principles of nucleoid organization have remained obscure due to the inability of conventional microscopy methods to retrieve structural information beyond the diffraction limit of light. Structured illumination microscopy has recently been shown to provide new levels of spatial details on bacterial chromosome organization by surpassing the diffraction limit. Its ease of use and fast 3D multicolor capabilities make it a method of choice for imaging fluorescently labeled specimens at the nanoscale. We describe a simple high-throughput method for imaging bacterial chromosomes using this technique.
细菌类核高度有序,但会通过转录、复制或分离等细胞过程进行动态重塑。由于传统显微镜方法无法获取超出光衍射极限的结构信息,类核组织的许多原理仍不清楚。最近研究表明,结构光照明显微镜通过突破衍射极限,能够提供细菌染色体组织新层次的空间细节。其易用性和快速的三维多色成像能力使其成为在纳米尺度对荧光标记样本成像的首选方法。我们描述了一种使用该技术对细菌染色体进行成像的简单高通量方法。