Journal Article Epublish 14 citations
PubMed: 34064730 DOI: 10.3390/ijms22105068
Bacteria Single-Cell and Photosensitizer Interaction Revealed by Quantitative Phase Imaging.
Int J Mol Sci 22(10)
Authors: Igor Buzalewicz, Agnieszka Ulatowska-Jarża, Aleksandra Kaczorowska, Marlena Gąsior-Głogowska, Halina Podbielska, Magdalena Karwańska, Alina Wieliczko, Anna K Matczuk, Katarzyna Kowal, Marta Kopaczyńska
Affiliations
- Department of Biomedical Engineering, Faculty of Fundamental Problems of Technology, Wrocław University of Science and Technology, 27 Wybrzeże S. Wyspiańskiego St. 50-370 Wrocław, Poland.
- Department of Epizootiology and Veterinary Administration with Clinic of Infectious Diseases, Wrocław University of Environmental and Life Sciences, 45 Grunwaldzki Square, 50-366 Wrocław, Poland.
- Department of Pathology, Division of Microbiology, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, 31 C.K. Norwida St. 51-375 Wrocław, Poland.
- Independent Researcher, Leeds 0113, UK.
Abstract
Quantifying 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 and 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.
MeSH
Cells, Photosensitizing Agents, Therapeutics, Bacteria, Quantitative Phase Imaging
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