Super-resolution fluorescence microscopy by stepwise optical saturation
Abstract
Super-resolution fluorescence microscopy is an important tool in biomedical research for its ability to discern features smaller than the diffraction limit. However, due to its difficult implementation and high cost, the super-resolution microscopy is not feasible in many applications. In this paper, we propose and demonstrate a saturation-based super-resolution fluorescence microscopy technique that can be easily implemented and requires neither additional hardware nor complex post-processing. The method is based on the principle of stepwise optical saturation (SOS), where M steps of raw fluorescence images are linearly combined to generate an image with a √M-fold increase in resolution compared with conventional diffraction-limited images. For example, linearly combining (scaling and subtracting) two images obtained at regular powers extends the resolution by a factor of 1.4 beyond the diffraction limit. The resolution improvement in SOS microscopy is theoretically infinite but practically is limited by the signal-to-noise ratio. We perform simulations and experimentally demonstrate super-resolution microscopy with both one-photon (confocal) and multiphoton excitation fluorescence. We show that with the multiphoton modality, the SOS microscopy can provide super-resolution imaging deep in scattering samples.
Yide Zhang
NIH K99 Postdoctoral Fellow
My research is interdisciplinary and focused on developing new types of optical imaging techniques that could advance the work of other researchers and medical personnel in a wide variety of fields. Currently, I am developing next-generation photoacoustic and ultrafast imaging techniques that can observe biological and physical phenomena that are too fast to be imaged with existing methods. The observation of the ultrafast phenomena could provide a better understanding of the fundamentals of life and physical sciences. I am also developing novel quantum imaging approaches that can investigate biological organisms with an imaging performance that cannot be achieved using classical optical imaging. In my free time, I enjoy cooking, hiking, cycling, and traveling.