Imaging of Bacterial Chromosome Organization by 3D Super-Resolution Microscopy

Methods Mol Biol. 2017:1624:253-268. doi: 10.1007/978-1-4939-7098-8_19.

Abstract

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.

Keywords: Bacterial chromosome; High-throughput; Nucleoid; Structured illumination microscopy; Super-resolution microscopy.

MeSH terms

  • Bacillus subtilis / genetics
  • Chromosomes, Bacterial / chemistry*
  • DNA Replication
  • Escherichia coli / genetics
  • Imaging, Three-Dimensional / methods*
  • Microscopy, Fluorescence