Photo-oxidation and Rebinding Kinetics of Bilirubin-bound UnaG for Super-resolution Imaging
124th Summer Symposium of KCS-Physical Chemistry Division
Single-molecule localization method is commonly used to construct a super-resolution image or to track a molecule of interest. The fitting accuracy of PSF to the image of fluorescence emission from a single molecule, which is the most frequently used method to find a molecule’s position, highly depends on the intensity of fluorescence and suffers from pixel-to-pixel variation of gain and noise of the camera. Thus the spatial resolution of single-molecule localization microscopy usually exceeds 10 nm. In this study, we developed a novel approach to localize a single molecule by illuminating it with a stripe-patterned light. When we modulated the phase of the illumination pattern, the total amount of emitted photons from a single molecule varied in proportion to the excitation intensity on the exact position of the molecule. Since the noisy features from pixel-to-pixel variation should be averaged during integrating the photons, our approach can provide better spatial resolution than conventional methods, especially under a low signal-to-noise ration condition.
References
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