Iterative Update Super-resolution Microscopic Imaging Method of Stripe Illumination in Fourier Domain Based on Total Internal Reflection

Abstract

The invention discloses a fringe illumination Fourier domain iteration updating super-resolution microscopic imaging method based on total internal reflection. The method comprises the following steps: splitting a parallel illumination laser beam into two parallel beams with equal intensity and consistent polarization direction, and exciting two oppositely propagated evanescent waves to interfereso as to generate a fine evanescent wave fringe illumination fluorescent sample; receiving a fluorescence signal on the imaging image surface by using a detector to obtain a low-resolution image mixedwith high-frequency and low-frequency information of the fluorescence sample; changing the spatial displacement and direction of the evanescent wave illumination fringes for multiple times, and shooting the fluorescence signal modulated by the fringe intensity again to obtain a series of low-resolution images mixed with high-frequency and low-frequency information of the fluorescence sample as original images; and finally, carrying out Fourier domain iteration updating processing on the original image, and carrying out continuous iteration to finally reconstruct a super-resolution image of the fluorescent sample. According to the method, the transverse resolution of about 100 nm can be achieved, the background level can be reduced, the contrast ratio can be improved, the unknown aberration of the system can be corrected, and in-vivo imaging can be achieved.

Description

technical field [0001] The invention relates to the field of optical super-resolution microscopic imaging, in particular to a method for iteratively updating super-resolution microscopic imaging in the Fourier domain of fringe illumination based on total internal reflection. Background technique [0002] Fluorescence microscopy plays a pivotal role in the application of biological research and clinical diagnosis due to its non-invasive characteristics. However, due to the existence of the diffraction limit, the resolution of optical microscopy usually cannot exceed 200nm. In the past few decades, many techniques have been invented to break through this diffraction limit. Among these microscopy techniques, structured illumination microscopy (SIM) is a powerful tool in biomedical imaging because the The method can provide high temporal and spatial resolution, and can achieve video-rate imaging speed. Although SIM can only achieve a two-fold improvement in resolution, it requ...

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Problems solved by technology

Other methods such as Photoactivated Localization Microscopy (PALM) and Stochastic Light Reconstruction Microscopy (STORM) are much slower than SIM for large fields of view, and they are difficult to reconstruct a super-resolution image. , requires thousands of original images, which makes it difficult to apply this method to in vivo imaging

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