English

Super-resolution fluorescence microscopy utilizes fluorescent probes and specific excitation and emission programs

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2024-01-23 14:07:18

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Super-resolution fluorescence microscopy surpasses the diffraction limit of what used to be a barrier by using fluorescent probes and specific excitation and emission programs. Most SR technologies heavily rely on image computation and processing to retrieve SR information. However, factors such as fluorescence group photophysics, chemical environment of the sample, and optical settings may cause noise and distortion in the original image, which may affect the quality of the final SR image.

Therefore, for SR microscope developers and users, having a reliable method to quantify reconstruction quality is crucial. Due to the improved distinguishability of SR imaging, it is necessary to conduct a thorough evaluation, but existing tools are often insufficient when the resolution of the authorities changes within the field of view.

In a recent study published in Light: Science and Applications, a group of scientists introduced a new method called rolling Fourier ring correlation. This method helps to directly represent resolution heterogeneity in the super-resolution domain, achieving unparalleled SR scale mapping and easily associating resolution mapping with SR content. In addition, the team also improved the resolution scaling error map to achieve more accurate system error estimation. This is combined with rFRC to create a combination technique called PANEL, which focuses on accurately locating low reliability areas from SR images.

Scientists have successfully applied PANEL to various imaging methods, including single molecule localization microscopy, super-resolution radial wave, structural illumination microscopy, and deconvolution methods, verifying the effectiveness and stability of its quantitative spectra. PANEL can be used to improve SR images. For example, it has been effectively used to fuse SMLM images reconstructed through various algorithms, providing high-quality SR images.

Source: Laser Net

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