Chapter title |
Imaging of Transcription and Replication in the Bacterial Chromosome with Multicolor Three-Dimensional Superresolution Structured Illumination Microscopy
|
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Chapter number | 7 |
Book title |
Bacterial Chromatin
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8675-0_7 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8674-3, 978-1-4939-8675-0
|
Authors |
Martin, Carmen Mata, Cagliero, Cedric, Sun, Zhe, Chen, De, Jin, Ding Jun, Carmen Mata Martin, Cedric Cagliero, Zhe Sun, De Chen, Ding Jun Jin |
Abstract |
Superresolution imaging technology has contributed to our understanding of the subnucleoid organization in E. coli cells. Multicolor superresolution images revealing "bacterial nucleolus-like structure or organization," "nucleolus-like compartmentalization of the transcription factories," and "spatial segregation of the transcription and replication machineries" have enhanced our understanding of the dynamic landscape of the bacterial chromatin. This chapter provides a brief introduction into multicolor three-dimensional superresolution structured illumination microscopy (3D-SIM) used to study the spatial organization of the transcription machinery and its spatial relationship with replisomes from a microbiological research perspective. In addition to a detailed protocol, practical considerations are discussed in relation to (1) sampling and treatment of cells containing fluorescent fusion proteins, (2) imaging the transcription and replication machineries at single-cell levels, (3) performing imaging experiments to capture the spatial organization of the transcription machinery and the nucleoid, and (4) image acquisition and analysis. |
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Unknown | 3 | 30% |