Method and system for rapidly three-dimensionally microimaging large sample

Abstract

The invention relates to a method and system for rapidly three-dimensionally microimaging a large sample. The method comprises the following steps: 1, a sample is embedded in a sample block and is fixed on a precise three-dimensional electric translation platform; 2, a digital micro mirror array works in a binary modulation mode, and is used for carrying out binary modulation on illuminating light to form structure striations, a structured light microscope collects an image of the surface layer of the sample within a depth of 1-100micrometers; 3, completing the imaging of the inner surface layer of an imaging region, removing an imaged part of the surface layer of the sample block by using a diamond or hard alloy tool; and 4, repeating the steps 2 and 3 until a data acquiring task is completed. According to the invention, the digital micro mirror array works in the binary modulation mode, the imaging speed is effectively increased; and with an exposure synchronization method of the digital micro mirror array and a surface array camera, the sample can be irradiated by exciting light only when a surface array camera is exposed, the exposure quantity of the sample is further reduced, and the negative effects of a light bleaching effect to imaging is avoided.

Description

technical field [0001] The present invention relates to a microscopic imaging method and system, in particular to a microscopic optical imaging method and system based on structured light three-dimensional tomography, which are suitable for large samples. Background technique [0002] In the field of optical microscopy, people have been pursuing the acquisition of three-dimensional detail information of samples in a larger imaging area. However, this goal is restricted by three factors. First, the existence of scattered light will lead to the generation of background light. As a result, traditional microscope imaging will not only obtain information on the focal plane of the objective lens, but also superimpose blurred background from other areas. Obtain the three-dimensional tomographic image of the sample; the second is limited by the field of view and imaging speed of the microscopic objective lens, it usually takes dozens of days for microscopic imaging of centimeter-sca...

AI Technical Summary

Problems solved by technology

Although the switching speed of a single micromirror in a digital micromirror array can reach tens of thousands of hertz, this grayscale modulation method limits its effective modulation speed to tens of hertz.

And the micro-mirrors are constantly flipped, so that even at the brightest gray scale, the gray-scale modulated micro-mirror array cannot reflect all the illumination light to the target area, and the utilization rate of illumination light is low, which reduces the structured light illumination pattern. The modulation depth affects the imaging speed and imaging effect

[0004] To sum up, the existing 3D microscopy imaging methods and systems for large samples have the following disadvantages: (1) The imaging speed is slow. For structured light microscopy, the imaging speed is mainly limited by the effective modulation frequency of the modulator; (2) ) Photobleaching affects image quality

These problems limit the development and application of such methods in practical applications, so it is necessary to develop a fast 3D microscopic imaging method and system suitable for large samples

Images