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Virtual Modulation Based High-Throughput Fluorescence Fluctuation Super-Resolution Imaging
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DOI：10.1021/acsphotonics.5c02613.png)
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Abstract
En 中文
Super-resolution optical fluctuation imaging (SOFI) has shown substantial potential in biological imaging due to its relaxed requirements for fluorophores. However, its application remains constrained by the requirement for a large number of frames and limited resolution enhancement. Here, we propose a virtual modulation based high-throughput fluorescence fluctuation imaging method, termed VSOFI. Leveraging the sectioning and high-frequency detection capabilities of array detection, VSOFI combines virtual modulation and iterative demodulation, achieving a 2-fold lateral resolution improvement and a 25-fold imaging efficiency increase with only 20 frames. Simulation and experimental results confirm that VSOFI maintains robust performance across various labeling densities and Ton/Toff contrasts. The method reliably resolves fine structures in both single- and dual-color imaging, and enables large-field visualization of microtubules in cell populations. As an easy-to-implement fluctuation imaging approach, VSOFI significantly broadens the applicability of SOFI and facilitates its integration into multimodal array-detection-based imaging systems.
Keywords:
super-resolution optical fluctuation imaging (SOFI)
array detection
virtual modulation
high-throughput