The rise of super-resolution fluorescence microscopy over the past decade has drastically improved the resolution of light microscopy to ∼10 nm. Among these methods, stochastic optical reconstruction microscopy (STORM) can be used to achieve sub-diffraction-limit resolution by sequentially imaging and localizing individual photoswitchable fluorophores. It is known that the switching rate of photoswitchable fluorophores depends on the laser power—a high laser power being required for the enhancement of imaging resolution. However, such high laser power usually results in photobleaching and high photo-toxicity effects, making it harmful to power-sensitive specimens, including biological samples. In this study, we attempted to overcome this problem by improving the STORM resolution at a lower laser power. Through the quantitative analysis of the photoswitching behavior of single fluorophores under different laser power conditions, we developed a new approach to achieve super-resolution fluorescence images at a laser power 10 times lower than had previously been reported. This approach is expected to play an increasingly significant role in super-resolution imaging of power-sensitive samples, such as live cells.

Development of a new approach for ultralow-laser-power super-resolution fluorescence imaging

ACS Spring 2022

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