微流控芯片与基质辅助激光解析电离质谱联用进行单细胞脂质组学分析

Lipids are one of important biological macromolecules in animals, possessing a variety of important biological functions. The traditional analysis of cell populations often ignores differences between individual cells, and it cannot truely reflect the physiological state of the cell. Therefore, it is essential to perform lipidomics study in single cells. However, the conventional analytical methods are difficult for single cell manipulation, and the trace amounts of metabolites in single cells are easy to loss during the sample preparation process. This research project is willing to design and optimize the structure and size of microchannels, and applying the fluid control technology to manipulate and immobilize single cells to form single cell microarrays; Optimization of different matrix and matrix combinations to improve the lipids detection sentivity of MALDI mass spectrometry; Exploring the optimal conditions for cell culture and drug stimulation of single cell microarrays on microfluidic device. Establishment of microfluidic device and MALDI mass spectrometry combination system for tumor single-cell lipidomics analysis to acquire significant difference of metabolites; The structural identification and biological validation of these metabolites, and discovering new lipid markers. The research project can monitor individual difference information between cell populations, which is very helpful for early diagnosis and monitoring of disease progression. It is very significant higher academic and clinical application values.

脂质是动物体内重要的生物大分子之一，具有多种重要生物学功能。传统的细胞群体分析，往往忽略了细胞个体间的差异，不能真实反映细胞的生理状态，所以，进行单细胞脂质组学研究至关重要。但是常规分析方法难于操纵单细胞，且样品预处理过程会导致单细胞内微量代谢物的部分丢失。本项目拟通过设计和优化微流控芯片的微通道结构与尺寸，利用流体操控技术进行操纵和固定并形成单细胞微阵列；优化不同基质和基质组合，提高MALDI质谱的脂质分析灵敏度；探索在微流控芯片上进行单细胞微阵列细胞培养和药物刺激的最佳条件；建立微流控芯片与MALDI质谱联用技术，进行肿瘤单细胞脂质组学分析，获得差异性代谢物；对差异物进行结构鉴定和生物学验证，发现新的脂质标志物。该项目研究成果可以监测群体细胞中个体间的差异信息，有助于疾病的早期诊断和疾病进程的监测，具有十分重要的学术意义和临床应用价值。

脂质是动物体内重要的生物大分子之一，具有多种重要生物学功能。传统的细胞群体分析，往往忽略了细胞个体间的差异，不能真实反映细胞的生理状态，所以，进行单细胞脂质组学研究至关重要。但是常规分析方法难于操纵单细胞，且样品预处理过程会导致单细胞内微量代谢物的部分丢失。本项目通过设计和优化基于微坑和微接触印刷技术的微流控芯片装置，利用流体操控技术实现了单细胞的高效率捕获形成单细胞阵列；同时还设计了基于纳米材料的N,S-CD新型基质，提高 MALDI 质谱对脂类物质及其他内源性小分子检测的灵敏度；通过优化在微流控芯片上进行单细胞微阵列细胞培养等条件，获得了单细胞的微阵列结构；建立微流控芯片与 MALDI 质谱联用技术，以肿瘤细胞A549为模型进行肿瘤单细胞脂质组学分析，共检测到12种脂类代谢物；对差异物进行结构鉴定和生物学验证，发现了新的脂质标志物。该项目研究成果可以监测群体细胞中个体间的差异信息，有助于疾病的早期诊断和疾病进程的监测，具有十分重要的学术意义和临床应用价值。