Time-Lapse imaging of bactericidal effect on nanostructural surface

Ito, Tetsuo; Nakade, Kazuki; Jindai, Keisuke; Sagawa, Takashi; Kojima, Hiroaki; Shimizu, Tomohiro; Shingubara, Shoso

doi:10.1117/12.2514492

Cited by 1 publication

(2 citation statements)

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“…When the membrane damage occurs, characteristic EPS production, bending of nanopillars, and leaking of cytoplasm are observed. These postmembrane damage characteristics have recently been observed in HIM, TEM, SEM, and confocal studies. ,,,,− However, as these events occur rapidly and only at the cell surface, the images obtained using these techniques commonly show the bacteria flattened on the nanopillars, apparently due to membrane damage. It should be noted that the interaction between bacterial membranes and nanostructures cannot be resolved with these microscopic tools due to the limitation in the time scale or spatial resolution.…”

Section: Introductionmentioning

confidence: 85%

“…Recently, the interfaces where bacteria and natural nanotopographies interact have been observed at nanoscale resolution through the use of some of the most advanced microscopic techniques available to-date including helium ion microscopy (HIM), transmission electron microscopy (TEM), focused ion beam (FIB) techniques, and faster atomic force microscopy (AFM). ,,,− These approaches have furthered our understanding of bacteria–nanotopography interactions of natural surfaces. High-resolution TEM images of cross-sectioned interfaces revealed that the natural nanopillars under the bacteria are bent during the interaction and that dead bacteria appeared to have produced EPS …”

Section: Introductionmentioning

confidence: 99%

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Resolving Bio–Nano Interactions of E. coli Bacteria–Dragonfly Wing Interface with Helium Ion and 3D-Structured Illumination Microscopy to Understand Bacterial Death on Nanotopography

Bandara

Ballerin

Leppänen

et al. 2020

ACS Biomater. Sci. Eng.

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Obtaining a comprehensive understanding of the bactericidal mechanisms of natural nanotextured surfaces is crucial for the development of fabricated nanotextured surfaces with efficient bactericidal activity. However, the scale, nature, and speed of bacteria–nanotextured surface interactions make the characterization of the interaction a challenging task. There are currently several different opinions regarding the possible mechanisms by which bacterial membrane damage occurs upon interacting with nanotextured surfaces. Advanced imaging methods could clarify this by enabling visualization of the interaction. Charged particle microscopes can achieve the required nanoscale resolution but are limited to dry samples. In contrast, light-based methods enable the characterization of living (hydrated) samples but are limited by the resolution achievable. Here we utilized both helium ion microscopy (HIM) and 3D structured illumination microscopy (3D-SIM) techniques to understand the interaction of Gram-negative bacterial membranes with nanopillars such as those found on dragonfly wings. Helium ion microscopy enables cutting and imaging at nanoscale resolution, while 3D-SIM is a super-resolution optical microscopy technique that allows visualization of live, unfixed bacteria at ∼100 nm resolution. Upon bacteria–nanopillar interaction, the energy stored due to the bending of natural nanopillars was estimated and compared with fabricated vertically aligned carbon nanotubes. With the same deflection, shorter dragonfly wing nanopillars store slightly higher energy compared to carbon nanotubes. This indicates that fabricated surfaces may achieve similar bactericidal efficiency as dragonfly wings. This study reports in situ characterization of bacteria–nanopillar interactions in real-time close to its natural state. These microscopic approaches will help further understanding of bacterial membrane interactions with nanotextured surfaces and the bactericidal mechanisms of nanotopographies so that more efficient bactericidal nanotextured surfaces can be designed and fabricated, and their bacteria–nanotopography interactions can be assessed in situ.

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