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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
Subject categories Biophysical chemistry, Microbiology


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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