Wide-field super-resolution microscopic imaging method and system thereof

Abstract

The invention discloses a wide-field super-resolution microscopic imaging method and system. The method comprises the steps: carrying out the phase modulation of laser emitted by a light source through a spatial light modulator, and obtaining intensity-modulated light through a Fourier lens; enabling the intensity-modulated light to sequentially pass through a tube mirror, an excitation light filter, a beam splitter and an objective lens, and then exciting a sample to emit signal light; collecting the signal light through the objective lens, and transmitting the signal light to a detector through the beam splitter; and performing reconstruction processing according to the signal light structure patterns of different phases in at least three orientations to obtain a super-resolution image.The spatial light modulator modulates the laser into an excitation light pattern whose intensity satisfies sinusoidal power distribution and excites a fluorescence pattern so as to generate multistagefrequency spectrum aliasing. And higher-frequency sample information is obtained by demodulating and resetting the aliasing frequency spectrum. Under an unsaturated excitation condition, super-resolution structured light illumination obvious micro-imaging is realized, and the spatial resolution is determined by the harmonic order of stripe structured light.

Description

technical field [0001] The invention relates to the technical field of optical microscopic imaging, in particular to a wide-field super-resolution microscopic imaging method and system thereof. Background technique [0002] Fluorescence microscopy has been widely used in life science research due to its advantages of non-destructive, non-invasive, specific labeling and real-time dynamic imaging of living cells. However, due to the limitation of the optical diffraction limit, the highest resolution of general imaging can only reach about 200nm. In order to break through the limitation of the diffraction limit on the resolution of fluorescence microscopy, a series of novel super-resolution microscopy imaging methods have been proposed. [0003] The Hell research group proposed the stimulated emission depletion (STED) technology, which can greatly improve the resolution to a level much higher than the diffraction-limited resolution (tens of nanometers or even higher). However...

AI Technical Summary

Problems solved by technology

At present, in SIM technology, only SSIM can achieve higher resolution than linear SIM. However, since SSIM requires high optical power excitation light to perform fluorescence saturation excitation on the sample, resulting in large optical damage, this method is not feasible. Suitable for live cell imaging, unable to take advantage of SIM

Images