Multiphoton Microscopy for Bacterial Imaging: A Label-free Solution Resting on Endogenous Two-photon Fluorescence

Cédric Delmon, Erwan Ferrandon, Emilie Chouzenoux, Audrey Prorot, Sophie Alain, Claire Lefort

2022

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-photon 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.

Paper Citation

in Harvard Style

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 - Volume 1: PHOTOPTICS, ISBN 978-989-758-554-8, pages 76-83. DOI: 10.5220/0010979600003121

in Bibtex Style

@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 - Volume 1: PHOTOPTICS,},
year={2022},
pages={76-83},
publisher={SciTePress},
organization={INSTICC},
doi={10.5220/0010979600003121},
isbn={978-989-758-554-8},
}

in EndNote Style

TY - CONF

JO - Proceedings of the 10th International Conference on Photonics, Optics and Laser Technology - Volume 1: PHOTOPTICS,
TI - Multiphoton Microscopy for Bacterial Imaging: A Label-free Solution Resting on Endogenous Two-photon Fluorescence
SN - 978-989-758-554-8
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