Real-time imaging of photodynamic action in bacteria
| Autor: | Tim Maisch, Anita Gollmer, Ariane Felgentraeger, Cristina Flors |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Fluorescence-lifetime imaging microscopy Staphylococcus aureus Porphyrins medicine.medical_treatment General Physics and Astronomy Photodynamic therapy 010402 general chemistry Photochemistry 01 natural sciences General Biochemistry Genetics and Molecular Biology 03 medical and health sciences chemistry.chemical_compound Cytosol Phenothiazines Phenothiazine Fluorescence microscope medicine General Materials Science Photosensitizing Agents Singlet Oxygen Chemistry Singlet oxygen General Engineering General Chemistry Porphyrin Fluorescence 0104 chemical sciences Methylene Blue 030104 developmental biology Microscopy Fluorescence Methylene blue |
| Zdroj: | Repositorio Institucional del Instituto Madrileño de Estudios Avanzados en Nanociencia instname |
| Popis: | Fluorescence imaging studies of the processes leading to photodynamic inactivation of bacteria have been limited due to the small size of microorganisms as well as by the faint fluorescence of most photosensitizers. A versatile method based on highly-sensitive fluorescence microscopy is presented which allows to study, in real time, the incorporation of photosensitizers inside S. aureus upon photodynamic action. The method takes advantage of the fluorescence enhancement of phenothiazine and porphyrin photosensitizers upon entering the bacterial cytosol after the cell wall has been compromised. In combination with typical assays, such as the addition of specific enhancers of reactive oxygen species, it is possible to extract mechanistic information about the pathway of photodynamic damage at the single-cell level. Imaging experiments in deuterated buffer strongly support a Type-I mechanism for methylene blue and a very minor role of singlet oxygen. |
| Databáze: | OpenAIRE |
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