| Autor: |
Wenzel M; Bacterial Cell Biology, Swammerdam Institute for Life Sciences, University of Amsterdam, 1098 XH, Amsterdam, The Netherlands. wenzelm@chalmers.se.; Department of Medical Microbiology and Infection Control, Amsterdam University Medical Centers - Location VUMC, 1081 HZ, Amsterdam, The Netherlands. wenzelm@chalmers.se.; Chemical Biology, Department for Biology and Biological Engineering, Chalmers University of Technology, 412 96, Gothenburg, Sweden. wenzelm@chalmers.se., Dekker MP; Department of Clinical Genetics, Center for Neurogenomics and Cognitive Research (CNCR), Neuroscience Campus Amsterdam, Amsterdam University Medical Centers - Location VUMC, 1081 HZ, Amsterdam, The Netherlands., Wang B; Bacterial Cell Biology, Swammerdam Institute for Life Sciences, University of Amsterdam, 1098 XH, Amsterdam, The Netherlands., Burggraaf MJ; Department of Medical Microbiology and Infection Control, Amsterdam University Medical Centers - Location VUMC, 1081 HZ, Amsterdam, The Netherlands., Bitter W; Department of Medical Microbiology and Infection Control, Amsterdam University Medical Centers - Location VUMC, 1081 HZ, Amsterdam, The Netherlands.; Department of Molecular Cell Biology, Amsterdam Institute for Molecules, Medicines, and Systems, Faculty of Science, Vrije Universiteit Amsterdam, 1081 HZ, Amsterdam, The Netherlands., van Weering JRT; Department of Clinical Genetics, Center for Neurogenomics and Cognitive Research (CNCR), Neuroscience Campus Amsterdam, Amsterdam University Medical Centers - Location VUMC, 1081 HZ, Amsterdam, The Netherlands. jan.van.weering@vu.nl., Hamoen LW; Bacterial Cell Biology, Swammerdam Institute for Life Sciences, University of Amsterdam, 1098 XH, Amsterdam, The Netherlands. |
| Abstrakt: |
Transmission electron microscopy of cell sample sections is a popular technique in microbiology. Currently, ultrathin sectioning is done on resin-embedded cell pellets, which consumes milli- to deciliters of culture and results in sections of randomly orientated cells. This is problematic for rod-shaped bacteria and often precludes large-scale quantification of morphological phenotypes due to the lack of sufficient numbers of longitudinally cut cells. Here we report a flat embedding method that enables observation of thousands of longitudinally cut cells per single section and only requires microliter culture volumes. We successfully applied this technique to Bacillus subtilis, Escherichia coli, Mycobacterium bovis, and Acholeplasma laidlawii. To assess the potential of the technique to quantify morphological phenotypes, we monitored antibiotic-induced changes in B. subtilis cells. Surprisingly, we found that the ribosome inhibitor tetracycline causes membrane deformations. Further investigations showed that tetracycline disturbs membrane organization and localization of the peripheral membrane proteins MinD, MinC, and MreB. These observations are not the result of ribosome inhibition but constitute a secondary antibacterial activity of tetracycline that so far has defied discovery. |
