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The Expandables: Cracking the Staphylococcal Cell Wall for Expansion Microscopy.


ABSTRACT: 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.

SUBMITTER: Kunz TC 

PROVIDER: S-EPMC8008081 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature

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The Expandables: Cracking the Staphylococcal Cell Wall for Expansion Microscopy.

Kunz Tobias C TC   Rühling Marcel M   Moldovan Adriana A   Paprotka Kerstin K   Kozjak-Pavlovic Vera V   Rudel Thomas T   Fraunholz Martin M  

Frontiers in cellular and infection microbiology 20210316


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 <i>Chlamydia trachomatis</i>, <i>Simkania negevensis</i>, and <i>Neisseria gonorrhoeae</i>. Gram-positive bacteria have a rigid and thick cell wall that impedes classic expansion strategies. Here we developed an approach, which included a seri  ...[more]

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