Journal
METHODS AND APPLICATIONS IN FLUORESCENCE
Volume 3, Issue 1, Pages -Publisher
IOP PUBLISHING LTD
DOI: 10.1088/2050-6120/3/1/014005
Keywords
super-resolution fluorescence microscopy; dSTORM; PALM; RNA polymerase; PAINT; chromosome structure; E. coli
Categories
Funding
- German Science Foundation (DFG) [EXC115, SFB902]
- European Molecular biology Organization [ASTF 573-2013]
Ask authors/readers for more resources
We demonstrate correlative super-resolution PALM, PAINT and dSTORM imaging of RNA polymerase, membrane and chromosomal DNA in fixed E. coli. This protocol allows the combination of precise structural information of the nucleoid (dSTORM) with quantitative super-resolution imaging (PALM) of interacting proteins. The spatial distribution and organization of RNA polymerase and DNA are visualized in bacterial cells grown at doubling times of 25 or 44 min. We observe that RNA polymerase is concentrated at the edge of the highly structured nucleoid during fast growth, whereas it is found more evenly distributed during medium-fast growth. In both conditions, the nucleoid shows densely packed areas which appear to be inaccessible to RNA polymerase. This finding is confirmed by live-cell tracking of RNA polymerase and subsequent imaging of the respective nucleoids using a protocol for fast fixation on-the-slide.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available