A flat embedding method for transmission electron microscopy reveals an unknown mechanism of tetracycline

• DOI: 10.1038/s42003-021-01809-8
• Publication Date: 2021-03-08T11:03:05Z
• Authors: Michaela Wenzel; Marien P. Dekker; Biwen Wang; Maroeska J. Burgg...; Wilbert Bitter; Jan R. T. van Wee...; Leendert W. Hamoen
• Keywords: 0301 basic medicine; 570; Tissue Embedding; QH301-705.5; Cell Membrane; Membrane Proteins; Microtomy; Tetracycline; Article; Anti-Bacterial Agents; 3. Good health; 03 medical and health sciences; Bacterial Proteins; Microscopy, Electron, Transmission; Biology (General); Bacillus subtilis

Abstract Transmission electron microscopy of cell sample sections is a popular technique in microbiology. Currently, ultrathin sectioning done on resin-embedded pellets, which consumes milli- to deciliters culture and results randomly orientated cells. This problematic for rod-shaped bacteria often precludes large-scale quantification morphological phenotypes due the lack sufficient numbers longitudinally cut Here we report flat embedding method that enables observation thousands cells per single section only requires microliter volumes. We successfully applied this Bacillus subtilis , Escherichia coli, Mycobacterium bovis Acholeplasma laidlawii . To assess potential quantify phenotypes, monitored antibiotic-induced changes B. Surprisingly, found ribosome inhibitor tetracycline causes membrane deformations. Further investigations showed disturbs organization localization peripheral proteins MinD, MinC, MreB. These observations are not result inhibition but constitute secondary antibacterial activity so far has defied discovery.