Yi-Jen Sun,1 Fan Bai,2 An-Chi Luo,1 Xiang-Yu Zhuang,1 Tsai-Shun Lin,1 Yu-Cheng Sung,1 Yu-Ling Shih,3 and Chien-Jung 1
1Department of Physics and Graduate Institute of Biophysics, National Central University, Jhongli, Taiwan ROC
2Biomedical Pioneering Innovation Center (BIOPIC), School of Life Sciences, Peking University, Beijing, China
3Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan, ROC
Microbiology, Time-Lapse Microscopy
The dynamic assembly of the cell wall is key to the maintenance of cell shape during bacterial growth. Here, we present a method for the analysis of Escherichia coli cell wall growth at high spatial and temporal resolution, which is achieved by tracing the movement of fluorescently labeled cell wall-anchored flagellar motors. Using this method, we clearly identify the active and inert zones of cell wall growth during bacterial elongation. Within the active zone, the insertion of newly synthesized peptidoglycan occurs homogeneously in the axial direction without twisting of the cell body. Based on the measured parameters, we formulate a Bernoulli shift map model to predict the partitioning of cell wall-anchored proteins following cell division.
Alexa Fluor 594 was excited by a 580-nm LED light source (58 W/cm2) (pE4000, CoolLED, UK) and was observed through a 650-nm emission filter.
Product Associated Features
Each of the four LED channels of the pE-4000 offer individual irradiance control. In addition to the 256 wavelength combinations, this is a flexible system for time-lapse applications where photobleaching and photodamage can be minimised to enable extended analysis.
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