Abstract:

Objective To establish a high-resolution three-dimensional imaging system with the Z-axis resolution improvement by ultra-thin cutting technology for tissue precise structure analysis. Methods Tissue samples were embedded with LRWhite resin and polymerized, which were cut into continuous slices of 50 nm-1 000 nm with diamond knife. Thereby the Z-axis resolution of 50 nm-1 000 nm was obtained. The semi-thin cutting or ultra-thin cutting slices were stained with methylation blue and basic fuchsin or labeled with antibody for structure illustration by automatic microscope. The high-resolution three-dimensional tissue spatial information was obtained with two-dimensional stitching and three-dimensional reconstruction. Results Using the established highresolution microscopic imaging technology, the fine structure of human normal colon tissues and mouse liver were observed. In combination with staining and antibody labeling technology, the distribution of tubulin in mouse brain tissues was observed by high-resolution three-dimensional imaging. Conclusion The established high-resolution three-dimensional imaging technology is applicable to the precise spatial structure of all kinds of tissues, and provides a solution to the Z-axis resolution for ultra-high-resolution imaging technology.

Key words: ultrathin cutting, tissue, high resolution, microscopy, antibody labeling