基于同步辐射技术开展单细胞整合素三维分布及定量分析研究

Develop method to analyse the true distribution and expression of protein on a single cell is a new challenge for biological analysis. In this program, we select integrin as model, design molecular probe to specifically target the integrin on a single cell; Rely on X-ray synchrotron radiation X-ray spectroscopy (nano-CT), XRF techniques and molecular probes, to obtain the 3D imaging of integrin and fine structure of intact cell, simultaneously quantify the integrin expression; Further, base on the mass signal of probe, to carry out highly sensitive quantitative analysis of integrin expression on single-cell level; Compare with the fluorescence imaging and biological transmission electron microscopy techniques, to optimize the accuracy and sensitivity of the method; Understand the correlation of tumor cell invasion ability with cell structures and markers expression level; Establish nano-synchrotron radiation integrated analysis method, to study the other interested proteins and cancer markers, and provide reference data for clinical diagnosis and research.

发展单细胞水平蛋白质三维分布及定量分析方法是生物分析中的新挑战。本项目以单细胞上整合素为研究对象，通过特异性识别单细胞整合素的分子探针，利用同步辐射纳米三维成像及X射线谱学技术，获得单细胞水平整合素的三维分布以及完整细胞的三维结构，开展整合素的细胞分布及定量分析；进一步利用探针质谱信号，开展高灵敏的单细胞水平整合素质谱定量分析；并对比经典荧光成像、生物电镜技术，优化方法的准确性和灵敏度；研究细胞微结构及标志物在不同侵袭能力肿瘤细胞中的变化，分析其与肿瘤侵袭能力以及病情发展的相关性；建立纳米-同步辐射融合的分析技术，为单细胞水平其他肿瘤标志分子及蛋白分析提供新的分析方案，为临床诊断及研究提供参考数据。