The Expandables: Cracking the Staphylococcal Cell Wall for Expansion Microscopy

Frontiers in Cellular and Infection Microbiology

Kerstin Paprotka¹  Vera Kozjak-Pavlovic¹  Martin Fraunholz¹  Marcel Rühling¹  Adriana Moldovan¹  Tobias C. Kunz¹  Thomas Rudel¹ 

[1] Department of Microbiology, Julius-Maximilians-Universität Würzburg, Würzburg, Germany;

关键词: high-resolution imaging;    endosomes;    autophagosomes;    host-pathogen interaction;    expansion microscopy;    Staphylococcus aureus;   

DOI  :  10.3389/fcimb.2021.644750

来源: Frontiers

【 摘 要 】

Expansion Microscopy (ExM) is a novel tool improving the resolution of fluorescence microscopy by linking the sample into a hydrogel that gets physically expanded in water. Previously, we have used ExM to visualize the intracellular Gram-negative pathogens Chlamydia trachomatis, Simkania negevensis, and Neisseria gonorrhoeae. Gram-positive bacteria have a rigid and thick cell wall that impedes classic expansion strategies. Here we developed an approach, which included a series of enzymatic treatments resulting in isotropic 4× expansion of the Gram-positive pathogen Staphylococcus aureus. We further demonstrate the suitability of the technique for imaging of planktonic bacteria as well as endocytosed, intracellular bacteria at a spatial resolution of approximately 60 nm with conventional confocal laser scanning microscopy.

【 授权许可】

CC BY