Photobleaching and phototoxicity of KillerRed in tumor spheroids induced by continuous wave and pulsed laser illumination
| Autor: | Pavel Subochev, Marina V. Shirmanova, Vladislav A. Kamensky, Daria S. Kuznetsova, Ilya V. Turchin, Varvara V. Dudenkova, Boris E. Shakhov, Elena V. Zagaynova, Sergey Lukyanov |
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| Rok vydání: | 2014 |
| Předmět: |
Materials science
Cell Survival medicine.medical_treatment Green Fluorescent Proteins Cell Culture Techniques General Physics and Astronomy Uterine Cervical Neoplasms Photodynamic therapy Apoptosis General Biochemistry Genetics and Molecular Biology law.invention Necrosis law medicine Humans General Materials Science Photosensitizer Irradiation Photobleaching Tissue Scaffolds Lasers Optical Imaging General Engineering Pulse duration General Chemistry Genetic Therapy Phototherapy Laser Photochemotherapy Biophysics Continuous wave Female Phototoxicity HeLa Cells |
| Zdroj: | Journal of biophotonics. 8(11-12) |
| ISSN: | 1864-0648 |
| Popis: | The purpose of this study was to evaluate photobleaching of the genetically encoded photosensitizer KillerRed in tumor spheroids upon pulsed and continuous wave (CW) laser irradiation and to analyze the mechanisms of cancer cell death after the treatment. We observed the light-dose dependent mechanism of KillerRed photobleaching over a wide range of fluence rates. Loss of fluorescence was limited to 80% at light doses of 150 J/cm(2) and more. Based on the bleaching curves, six PDT regimes were applied for irradiation using CW and pulsed regimes at a power density of 160 mW/cm(2) and light doses of 140 J/cm(2) , 170 J/cm(2) and 200 J/cm(2). Irradiation of KillerRed-expressing spheroids in the pulsed mode (pulse duration 15 ns, pulse repetition rate 10 Hz) induced predominantly apoptotic cell death, while in the case of CW mode the cancer cells underwent necrosis. In general, these results improve our understanding of photobleaching mechanisms in GFP-like proteins and show the importance of appropriate selection of treatment mode for PDT with KillerRed. Representative fluorescence image of two KillerRed-expressing spheroids before and immediately after CW irradiation. |
| Databáze: | OpenAIRE |
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