基于光催化快速标记-富集技术的原位单细胞质谱代谢组定量分析系统的研究

为解决原位单细胞质谱分析（ambient single-cell mass spectrometry）中检测灵敏度低、检测覆盖范围窄以及原位定量困难等瓶颈问题，本项目基于光催化有机反应的快速高效性以及部分催化材料的富集能力，拟发展光催化标记平台引入原位液相萃取-电喷雾质谱系统中，实现代谢物的原位单细胞采样-光控快速标记富集-质谱检测。同时，基于采样动力学模型开发无需样品内加标的绝对定量校正方法。研究将着重于三点：一是光催化标记反应，包括光催化材料的掺杂和形貌改进以及高活性标记试剂的开发；二是光控快速富集，包括光照对代谢物在材料上吸附行为的影响以及加速解析的研究；三是单细胞采样动力学模型与动力学定量校正方法的建立。最终，项目将建立针对复杂代谢组的高灵敏度、高检测覆盖范围和强绝对定量能力的原位单细胞质谱分析平台，为解析细胞在生命活动和疾病过程中的个体差异提供更高效高通量且可靠的检测技术。

The low sensitivity, narrow detection coverage and poor quantitation capability are the bottle necks of current ambient single-cell mass spectrometry (MS) techniques. To address these issues, we propose to develop rapid and efficient photocatalytical labeling reactions and introduce them into ambient liquid extraction mass spectrometry system, and finally establish an ambient single cell sampling-light controlled rapid enrichment/labeling-MS platform for metabolomics based on the high efficiency and reaction rate of photocatalytical organic reaction and the enrichment capability of some photocatalytical nanomaterials. Moreover, the kinetic model for the sampling process will be established and used to develop the kinetic calibration method for in situ quantitation without spiking standards in samples. We would focus on three points in the project: First is the photocatalytical labeling method, including doping/hybridization and morphology optimization of photocatalytical materials immobilized in capillary, and development of highly active labeling reagents; second is the light-controlled enrichment of analytes on the photocatalytical materials, including the investigation of the effect of light on the adsorption behavior and the desorption rate of analytes on the materials; third is the establishment of kinetic calibration quantitation method based on the mass transfer kinetic model in single-cell sampling. Finally, a sensitive and quantitative ambient single-cell MS analysis platform with enhanced detection coverage will be established for metabolomics. This study would provide a more efficient, high throughput and reliable analytical technique for research on cell heterogeneity in living and disease processes.