螺吡喃衍生物修饰的光控纳喷毛细管探针用于单细胞质谱分析的研究

In order to avoid the limitations of cell homogenization study, it is very important to qualitatively and quantitatively determine the compositions and contents which are tightly related with bioactivities in single cell, such as amino acids, sugars, lipids, lactate and so on. To overcome the obvious difficulties in single cell analysis, such as small cell sap volume (fL to pL level), low material content, big difference in concentration and ionization ability which is easy to miss the information of compounds with weak ionization ability, as well as inaccurate calculation in sample volume, we propose a novel method to rapidly examine the compositions in single cell using a photo-controlled, gold-plated capillary probe coupled with mass spectrometry. The probe has multiple functions: The equal inner diameter tip can insert into the single cell and pL volume of samples from the cell sap could be precisely pipetted by a electroosmotical pump; The inner wall of the probe modified by spiropyran could be selectively absorb the hydrophilic compounds or hydrophobic compounds under UV or Visible light condition, which can examine the compounds with nonpolar, weak ionization ability or polar, strong ionization ability respectively, thus reducing the background and increasing the sensitivity. This novel and efficient mass spectrometry method will not only provide a powerful tool to study the compositions and minor changes in single cells, but also have potential applications in the fields of biology and medicine.

为了避免细胞均化研究的局限性，在单细胞水平上对细胞内与各种生理活动密切相关的多种物质(如氨基酸、糖类、脂质、乳酸等)进行定性与定量分析就显得至关重要。然而单细胞细胞液体积小，通常在fL-pL级别，无法精确的计算取样体积；单细胞内物质含量低、浓度差异大、电离能力不同、容易丢失部分物质信息。针对这两个科学问题，本项目提出构建一种新颖的光控纳喷毛细管探针−质谱检测系统用于单细胞内小分子成分的检测。光控纳喷毛细管探针将单细胞取样与质谱检测集于一体，其等内径尖端可以扎取单细胞，并在电渗泵的驱动下实现皮升级样品的精确定量取样；螺吡喃衍生物修饰化的内壁在可见/紫外光条件下，可以实现对非极性、弱电离能力的成分和极性、强电离能力的成分的分别检测，从而减小背景干扰，提高检测的灵敏度。本项目的实施为质谱研究单细胞的组成成分及细胞内物质的微小变化提供了新策略，在生物医学研究领域具有潜在的应用价值。

为了避免细胞均化研究的局限性，在单细胞水平上对细胞内各种与生命活动密切相关的物质（如氨基酸类、糖类、脂质、乳酸等）进行定性与定量分析就显得至关重要。然而单细胞细胞液体积小，通常在fL-pL级别，无法精确的计算取样体积；单细胞内物质含量低、浓度差异大、电离能力不同、容易丢失部分物质信息。针对这两个科学问题，本项目提出构建一种新颖的螺吡喃光控纳喷毛细管探针质谱检测系统用于单细胞小分子成分的检测。光控纳喷毛细管探针将单细胞取样与质谱检测集于一体，其等内径尖端可以扎取单细胞后直接进行质谱检测；螺吡喃衍生物修饰化的内壁在可见/紫外光条件下，可以分别实现对非极性、弱电离能力的成分和极性、强电离能力的成分的检测，从而减小背景干扰，提高检测的灵敏度。本项目的实施为质谱研究单细胞的组成成分及细胞内物质的微小变化提供了新策略，在生物医学研究领域具有潜在的应用价值。