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Total Internal Reflection Fluorescence (TIRF) Microscopy is widely used in cell biology to monitor dynamic biomolecular events occurring at the plasma membrane in living cells such as virus assemblies and particle budding. However, these studies are limited by the weak fluorescence signal and background noise degrading the spatial resolution. We present highly sensitive fluorescent TIRF imaging using classical glass coverslips coated with a resonant multi-dielectric thin film. Such dielectric multilayer is optimized to generate a large field enhancement under TIR illumination at the free interface. However, dielectric materials usually have low imaginary indices that are not experimentally measurable and introducing large discrepancies with the theory. We tackled the k issue by adjusting the oxygen level within the last thin layer to fully control its absorption. We then qualify such dielectric stack resonance in TIRFM configuration for fluorescent viruses imaging.
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