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Exploring photoinactivation of microbial biofilms using laser scanning microscopy and confined two-photon excitation.

Journal article
Authors Hanna Thomsen
Fabrice Graf
Anne Farewell
Marica B Ericson
Published in Journal of biophotonics
Volume 11
Issue 10
ISSN 1864-0648
Publication year 2018
Published at Department of Chemistry and Molecular Biology
Centre for antibiotic resistance research, CARe
Language en
Links dx.doi.org/10.1002/jbio.201800018
www.ncbi.nlm.nih.gov/entrez/query.f...
Subject categories Biophysical chemistry, Microbiology

Abstract

One pertinent complication in bacterial infection is the growth of biofilms, i.e., communities of surface-adhered bacteria resilient to antibiotics. Photodynamic inactivation has been proposed as an alternative to antibiotic treatment; however, novel techniques complementing standard efficacy measures are required. Herein, we present an approach employing multiphoton microscopy complemented with Airyscan super-resolution microscopy, to visualize the distribution of curcumin in Staphylococcus epidermidis biofilms. The effects of complexation of curcumin with hydroxypropyl-γ-cyclodextrin (HPγCD) were studied. It was shown that HPγCD-curcumin demonstrated higher bioavailability in the biofilms compared to curcumin, without affecting the subcellular uptake. Spectral quantification following photodynamic inactivation demonstrates a method for monitoring elimination of biofilms in real time using noninvasive 3D imaging. Additionally, spatially confined two-photon inactivation was demonstrated for the first time in biofilms. These results support the feasibility of advanced optical microscopy as a sensitive tool for evaluating treatment efficacy in biofilms towards improved mechanistic studies of photodynamic inactivation. This article is protected by copyright. All rights reserved.

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