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Authors: Cédric Delmon 1 ; Erwan Ferrandon 2 ; Emilie Chouzenoux 3 ; Audrey Prorot 1 ; Sophie Alain 4 and Claire Lefort 2

Affiliations: 1 Peirene, GRESE (EA 4330), University of Limoges, Limoges, France ; 2 XLIM Research Institute, UMR CNRS 7252, University of Limoges, Limoges, France ; 3 Center for Visual Computing, CentraleSupélec, Inria Saclay, Université Paris-Saclay, Gif-sur-Yvette, France ; 4 RESINFIT, UMR INSERM 1092, University of Limoges, Bacteriology-Virology-Hygiene Department, University Hospital Center, France

Keyword(s): Multiphoton Microscopy, Bacterial Imaging, Spectral Characterization, Two-photon Excitation, Fluorescence Emission, Metabolic Indicators, Point-Spread-Function, Computational Restoration, Block Distributed Majorize-minimize Memory Gradient.

Abstract: We demonstrate the interest of multiphoton microscopy (MPM) for imaging bacteria without any labelling process. Six families of bacteria are tested: Escherichia coli, Staphylococcus epidermidis, Proteus vulgaris, Pseudomonas fluorescens, Bacillus subtilis and Clostridium perfringens. For each of these bacteria, the image of a cell is recorded through a multiphoton microscope thus revealing the 3D shape of these bacteria. For the first time, the images of such bacteria are recorded without any labelling solution. A protocol of controlling the image produced is led thanks to a standard staining protocol with carboxy fluorescein diacetate (CFDA) for E. Coli and Staphylococcus epidermidis. Similar object shapes with or without labelling are produced, thus validating the label-free images generated by MPM. Then, the two-photon excitation spectra are measured for each of these bacteriaand the emission spectra delivered by E. Coli and Bacillus subtilis are shown. The origin of the two-photo n fluorescence (TPF) emission of the bacteria thanks to the nonlinear imaging solution is discussed regarding to the TPF excitation and emission spectra of metabolic indicators. (More)

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Paper citation in several formats:
Delmon, C.; Ferrandon, E.; Chouzenoux, E.; Prorot, A.; Alain, S. and Lefort, C. (2022). Multiphoton Microscopy for Bacterial Imaging: A Label-free Solution Resting on Endogenous Two-photon Fluorescence. In Proceedings of the 10th International Conference on Photonics, Optics and Laser Technology - PHOTOPTICS; ISBN 978-989-758-554-8; ISSN 2184-4364, SciTePress, pages 76-83. DOI: 10.5220/0010979600003121

@conference{photoptics22,
author={Cédric Delmon. and Erwan Ferrandon. and Emilie Chouzenoux. and Audrey Prorot. and Sophie Alain. and Claire Lefort.},
title={Multiphoton Microscopy for Bacterial Imaging: A Label-free Solution Resting on Endogenous Two-photon Fluorescence},
booktitle={Proceedings of the 10th International Conference on Photonics, Optics and Laser Technology - PHOTOPTICS},
year={2022},
pages={76-83},
publisher={SciTePress},
organization={INSTICC},
doi={10.5220/0010979600003121},
isbn={978-989-758-554-8},
issn={2184-4364},
}

TY - CONF

JO - Proceedings of the 10th International Conference on Photonics, Optics and Laser Technology - PHOTOPTICS
TI - Multiphoton Microscopy for Bacterial Imaging: A Label-free Solution Resting on Endogenous Two-photon Fluorescence
SN - 978-989-758-554-8
IS - 2184-4364
AU - Delmon, C.
AU - Ferrandon, E.
AU - Chouzenoux, E.
AU - Prorot, A.
AU - Alain, S.
AU - Lefort, C.
PY - 2022
SP - 76
EP - 83
DO - 10.5220/0010979600003121
PB - SciTePress