Image scanning microscopic imaging method and device based on double-microlens array

Abstract

The invention relates to an image scanning microscopic imaging method and device based on a double-microlens array, and belongs to the technical field of optical microscopic measurement. According tothe invention, an array type multi-focus generated by the micro-lens array is used for illuminating a sample; the imaging speed is improved, the influence of out-of-focus fluorescence is eliminated through the pinhole array, each focusing light beam is scaled by 1 / 2 through the second micro lens array, all-optical implementation of subsequent digital information processing of traditional image scanning microscopy is achieved, it is guaranteed that the resolution ratio is improved by two times, and meanwhile the problem that a traditional image scanning microscopy system is low in imaging speedis solved. The laser beam reaches the first micro-lens array after being expanded to generate an array type illumination beam, and after the array type illumination beam is emitted to the surface ofa sample, the objective lens collects multi-focus fluorescence from the sample and emits the multi-focus fluorescence to the pinhole array to eliminate the influence of out-of-focus fluorescence. Thesecond micro-lens array locally shrinks each focus point, meanwhile, the original focusing direction of the light beam is kept, and the zoomed fluorescence focus is imaged to the sCMOS camera under the deflection effect of the scanning galvanometer.

Description

technical field [0001] An image scanning microscopic imaging method and device based on a double microlens array belongs to the field of optical microscopic measurement, and mainly relates to a method and a device for non-contact rapid imaging of fine structures of biological samples based on microstructured optical elements. Background technique [0002] Modern fluorescence microscopy combines imaging speed, molecular specificity, and contrast to visualize the cellular imaging process, but diffraction limits the resolution of wide-field microfluorescence microscopy, while super-resolution imaging breaks through the traditional diffraction limit, but in There is a decrease in speed, imaging time and field of view. For example, single-molecule imaging counting and stimulated emission depletion microscopy enable cells with a spatial resolution of less than 100 nm, but imaging speeds are limited to 0.01-1 Hz. [0003] Compared with single-molecule imaging counting and stimulat...

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Problems solved by technology

[0006] In order to solve the problems of slow imaging speed and complex subsequent reconstruction algorithm of image scanning microscopic imaging technology, the present invention designs an image scanning microscopic imaging method and device based on a double microlens array. The excitation beam is generated by the first microlens array The array excitation beam realizes multi-focus simultaneous scanning of the sample, improves the scanning speed, and uses the second scanning lens to zoom the original image to 1 / 2 of the original image, realizes optical image processing, saves subsequent digital image processing operations, and reduces rebuild time

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