Bacteria Single-Cell and Photosensitizer Interaction Revealed by Quantitative Phase Imaging
Igor Buzalewicz , Agnieszka Ulatowska-Jarża , Aleksandra Kaczorowska , Marlena Gąsior-Głogowska , Halina Podbielska , Magdalena Karwańska , Alina Wieliczko , Anna Matczuk , Katarzyna Kowal , Marta Kopaczyńska
AbstractQuantifying changes in bacteria cells in the presence of antibacterial treatment is one of the main challenges facing contemporary medicine; it is a challenge that is relevant for tackling issues pertaining to bacterial biofilm formation that substantially decreases susceptibility to biocidal agents. Three-dimensional label-free imaging and quantitative analysis of bacteria–photosensitizer interactions, crucial for antimicrobial photodynamic therapy, is still limited due to the use of conventional imaging techniques. We present a new method for investigating the alterations in living cells and quantitatively analyzing the process of bacteria photodynamic inactivation. Digital holographic tomography (DHT) was used for in situ examination of the response of Escherichia coli and Staphylococcus aureus to the accumulation of the photosensitizers immobilized in the copolymer revealed by the changes in the 3D refractive index distributions of single cells. Obtained results were confirmed by confocal microscopy and statistical analysis. We demonstrated that DHT enables real-time characterization of the subcellular structures, the biophysical processes, and the induced local changes of the intracellular density in a label-free manner and at sub-micrometer spatial resolution.
|Journal series||International Journal of Molecular Sciences, ISSN 1422-0067, (N/A 140 pkt)|
|Publication size in sheets||1.15|
|Keywords in English||digital holographic tomography; photodynamic inactivation; single-cell bacteria|
|ASJC Classification||; ; ; ; ; ; ;|
|License||Journal (articles only); published final; ; with publication|
|Score||= 140.0, 17-05-2021, ArticleFromJournal|
|Publication indicators||: 2018 = 1.224; : 2018 = 4.183 (2) - 2018=4.331 (5)|
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