›› 2013, Vol. 33 ›› Issue (1): 121-.doi: 10.3969/j.issn.1674-8115.2013.01.025

• Technique and method • Previous Articles     Next Articles

Tissue precise spatial structure determination with ultra-thin cutting high-resolution imaging technology

LÜ|Chun-Mei1, MENG Jun1, LI Xiao-lin1, LI Xiao-wei1, GUO Yan1,2   

  1. 1.Key Laboratory of the Ministry of Education for Systems Biomedicine, Institute for Systems Biomedicine, Shanghai Jiaotong University, Shanghai 200240, China; 2.National Laboratory for Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai Jiaotong University, Shanghai 200032, China
  • Online:2013-01-28 Published:2013-02-06
  • Supported by:

    National Natural Science Foundation of China, 30900271, 60907044; National Natural Science Major Research Program, 91229108


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