Wide-field super resolution fluorescence microscopic imaging device

Abstract

The invention discloses a wide-field super resolution fluorescence microscopic imaging device which comprises a light source, a modulation unit, a dichroscope and an imaging unit, wherein the modulation unit modulates light beams emitted by the light source into two beams of polarized light p and two beams of polarized light s which can interfere with one another and is used for changing interference phase difference of the two sets of light beams; the two beams of polarized light p and the two beams of polarized light s form interference fringes on the surface of the dichroscope and reflected by the dichroscope to serve as structural speckle illumination light of an illumination sample, and the structural speckle illumination light has bright specks which are distributed in arrays; the imaging unit comprises a microobjective for projecting the structural speckle illumination light to the sample and a camera for conducting stimulated radiation fluorescent imaging on the sample.By means of the wide-field super resolution fluorescence microscopic imaging device, a wide imaging view field and a resolution ratio of super diffraction limit can be achieved at the same time, and frequency shift iteration of multiple images is conducted without depending on a structural light function, so that the super resolution ratio of images can be guaranteed quite well, and the signal noise ratio of images can be raised.

Description

technical field [0001] The invention relates to the field of optical super-resolution microscopy, in particular to a microscopic imaging device that takes into account both wide field of view and super-resolution. Background technique [0002] As a non-contact and non-destructive detection method, super-resolution fluorescence microscopy can reveal the dynamic and structural characteristics of intracellular molecular scale, and plays an extremely important role in the field of life sciences. [0003] According to the theory of Fourier optics, the traditional optical system is limited by its spatial-bandwidth product (SBP), and it is often difficult to balance super-resolution and wide field of view in a single imaging process. One of the bottlenecks in the development of imaging technology. [0004] At present, the relatively successful super-resolution fluorescence microscopy techniques mainly include the following: single-molecule fluorescence imaging (PALM and STORM), st...

AI Technical Summary

Problems solved by technology

STED superimposes a hollow spot on a Gaussian spot, which suppresses the fluorescent radiation around the Gaussian spot, and then realizes super-resolution imaging. However, its imaging adopts a point-by-point scanning method, which limits the imaging field of view. imaging, temporal resolution must be sacrificed

SIM and SSIM themselves use structured light to illuminate the sample, so they have a larger field of view, but the resolution of SIM can only break through the diffraction limit twice in theory, while the super-resolution image inversion process of SSIM largely relies on The correctness of the structured light function, but in the process of operation, affected by the actual system setting error, laser speckle, system noise and other factors, the structured light function is always different from the expected, which will affect the super-resolution inversion effect, and The signal-to-noise ratio of the entire image needs to be further improved

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