A method and device for super-resolution microscopy based on speckle illumination

Abstract

The invention discloses a super-resolution microscopy method based on speckle illumination. The method comprises the following steps: modulating laser beam, focusing the laser beam onto a sample to be tested, so as to form a speckle illumination pattern, and collecting fluorescence emitted by the sample to be tested, so as to obtain a fluorescence intensity image; changing the speckle illumination pattern, so as to obtain a plurality of fluorescence intensity images under different speckle illumination patterns; adding all the fluorescence intensity images together to obtain an image serving as a wide field image, and performing deconvolution on the wide field image, so as to obtain the initial estimation of the sample; calculating an initial illumination image through a gradient descent algorithm according to the obtained initial estimation; working out a sample image with higher resolution on the basis of an obtained object image and an initial illumination image through an FP algorithm; taking the worked out sample image as an estimated value of the sample, repeating iteration till iteration is accomplished, so as to obtain a super-resolution image. The invention further discloses a super-resolution microscopy device based on speckle illumination.

Description

technical field [0001] The invention belongs to the field of optical super-resolution microscopy, in particular to a super-resolution microscopy method and device based on speckle illumination. Background technique [0002] With the development of science and technology, people's demand for the microcosm is increasing, especially in life science and material science. In order to meet the exploration of nanoscale material information, many scholars are working on the research of improving the resolution. The super-resolution techniques that have been realized so far include: structured illumination microscopy (SIM), stimulated emission depletion (STED), stochastic optical reconstruction microscopy , STORM), photoactivated localization microscopy (PALM), etc. [0003] Among the above-mentioned super-resolution imaging technologies, the imaging speed of STORM and PALM is limited, which greatly limits the development of basic biological research work such as molecular dynamics...

AI Technical Summary

Problems solved by technology

At present, the optimization algorithm based on blind-SIM is PE-SIMS-PR, but when this covariance-based blind-SIM algorithm restores a grayscale image, the final result will cause the weak fluorescent signal to be lost, thus damaging the Information about the final image

[0006] Similarly, the FP (pattern-illuminated Fourier ptychography, piFP) algorithm of Sanban lighting does not require precise control of the lighting pattern, but this method needs to know the precise position of the lighting pattern movement, which also brings great challenges to the realization of this algorithm. some difficulties

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