Tissue and organ three-dimensional imaging and analysis method based on continuous slicing, multicolor fluorescence and three-dimensional reconstruction

Abstract

The invention discloses a tissue and organ three-dimensional imaging and analysis method based on continuous slicing, multicolor fluorescence and three-dimensional reconstruction. The method mainly comprises the following steps: strictly and continuously slicing a reconstructed object; alternately carrying out multi-color immunofluorescence staining in multiple combinations according to a standardprocess; carrying out multi-channel fluorescence scanning on all the immunofluorescence staining sections to obtain digital section images; aligning continuous slices according to fluorescence channel signals, and inserting various dyeing combinations for three-dimensional reconstruction to obtain a tissue and organ fluorescence three-dimensional model with cell-level resolution; and conducting STL modeling on the interested local area, and conducting various kinds of analysis. According to the method, a high-resolution tissue and organ fluorescence three-dimensional model capable of performing quantitative analysis can be constructed, and the tissue and organ three-dimensional form can be faithfully restored to the maximum extent; one or more fluorescence channels can be checked independently or in an overlapping manner; and STL modeling can be performed on the region of interest to complete spatial quantification and correlation analysis of various cells and functional markers.

Description

Technical field[0001]The invention relates to the field of tissue and organ stereo imaging analysis, in particular to a tissue and organ stereo imaging and analysis method based on continuous slices, multicolor fluorescence and three-dimensional reconstruction.Background technique[0002]Tissue and organ specimens are important research materials in the field of biomedical research, as well as key specimens for disease diagnosis. Tissue imaging is an important means of diagnosis and research on tissue specimens. Tissue imaging mainly includes staining and detection.[0003]Staining methods include dye staining (such as HE staining) and antibody-based staining. The latter is divided into immunohistochemical staining (IHC) and immunofluorescence staining (IF). Among them, compared with IHC, IF can detect multiple antigens at the same time, which is helpful for more accurate cell identity confirmation and functional characterization.[0004]The detection part is still based on the observatio...

AI Technical Summary

Problems solved by technology

In 2017, Yan Huanhuan et al. used continuous section IHC staining and layer-by-layer calibration for three-dimensional tissue reconstruction (application publication number: CN 106918484 A), and its shortcomings mainly include the single IHC detection index, and the formed three-dimensional model can reflect relative information. It is relatively limited, and the workload of layer-by-layer calibration of scanned images is large, resulting in high labor costs and relatively low clinical or scientific practical value of this method

In addition, although the three-dimensional scanning imaging technology based on tissue clearing staining microscope has the advantages of good tissue integrity, it also has disadvantages such as low model resolution, single staining markers, high hardware requirements, high cost, and difficulty in popularization.

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