Study protocol to determine wavelength, fluence, and irradiance activities of Os-4T:
- Lead in compound family (Os-4T) was applied to SK-MEL-28 melanoma cells in well-plates
- Well-by-well illuminations in triplicates with the ML8500 (37 ⁰C, 5% CO2) using 525 nm and 630 nm wavelengths. In first experiment irradiance was kept constant at 300 mW/cm2 and fluence was varied between 10-300 J/cm2, while in second experiment fluence was constant at 100 J/cm2 and irradiance varied 25-300 mW/cm2.
- Determine cell viability with Resazurin assay and read absorbance at 570 and 600 nm. Resazurin assay principle: The irreversible reaction of purple resazurin to pink-colored resorufin is proportional to aerobic respiration, indicating the amount of viable cells.
Illumination parameters significantly affected the phototoxicity of Os-4T. Green light (525 nm) was more effective than red light (630 nm) at lower fluences, while maximal potency was achieved at both wavelengths when using fluences higher than 100 J/cm2. Irradiance affected the potency with both red and green light that were more effective when lower irradiances were used.
Graphs from the original publication. Open-access content reproduced with permission from the contact author of the publication.
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