dSTORM: real-time subdiffraction-resolution fluorescence imaging with organic fluorophores

In the recent past, a variety of methods have been developed to circumvent the diffraction barrier of light which restricts optical resolution to about 200 nm in the image plane. Single-molecule based photoswitching microscopy such as direct stochastic optical reconstruction microscopy (dSTORM) has been successfully implemented for subdiffraction-resolution fluorescence imaging. The major drawback of this technique has been that the reconstruction of subdiffraction-resolution images requires substantially more time than the actual experiment and prevented real-time imaging. Here we present a new computational algorithm enabling subdiffraction-resolution fast imaging of cellular structures with ~20 nm optical resolution in less than 10 seconds.

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