Structure illumination super-resolution microscopy imaging system and imaging method thereof

Abstract

The invention discloses a structure illumination super-resolution microscopy imaging system and an imaging method thereof. The imaging system comprises an illumination light source, a rotating structure light generator, a first convergent lens, a light splitter, an objective lens, an object stage, a sample, a second convergent lens, a digital imaging device and a computer. An original image is processed through an imaging image reconstruction algorithm; super-resolution can be achieved only by rotating structured light stripes for four times without phase displacement; the imaging image reconstruction algorithm is based on frequency domain processing instead of non-spatial domain processing; a spectrum process of a super-resolution image of a sample is different from a traditional frequency domain method, structured light directions are not analyzed one by one and all directions are merged for analysis; in order to obtain traditional uniform 2-fold resolution improvement in various directions, the theoretical quantity of original images can be reduced to 4 from 9 of a traditional method; the quantity of the original images is further reduced to 3 and uniform 1.5fold resolution improvement in various direction can be theoretically obtained.

Description

technical field

[0001] The invention relates to optical microscopic imaging technology, in particular to a structured light illumination super-resolution microscopic imaging system and an imaging method thereof. Background technique

[0002] Super-resolution optical microscopy utilizes spatial or temporal modulation of fluorescence emission to achieve microscopic imaging that breaks through the optical diffraction limit. Structure Illumination Microscopy (SIM), based on wide-field fluorescence microscopy, uses specially modulated structured light to illuminate the sample, uses specific algorithms to extract high-frequency information from the modulated image, and breaks through diffraction. The limit of the limit.

[0003] Conventional reconstruction methods for 2D SIM require at least 9 original images. The reconstruction method proposed by Gustafsson [1] , take 3 different initial phases in each lighting direction value, obtain 3 original images, and obtain the high-f...

Images