Novel Platform for Quantitative Subcellular Resolution Imaging of Human Tissues Using Mass Spectrometry

一种新的人体组织亚细胞分辨率定量成像平台 质谱学 质谱学成像(MSI)是一种强大的技术，它实现了无标记空间 生物系统中不同类别生物分子的作图。因为它不需要 任何特殊的样品前处理，环境MSI特别吸引高通量 自动化成像应用程序。环境MSI实验的吞吐量通常是有限的 通过从表面进行固有的缓慢的微探针式采样，这是一个特征 许多化学成像方式的缺点。这个项目将结合几个高度 采用创新的方法来应对与高吞吐量、高 纳米螺旋解吸电喷雾用于脂类及其代谢物的环境MSI分析 电离(纳米-DESI)。纳米DESI是一种大气电离技术，它依赖于温和的 将组织切片中的分子局部液萃取到流动的受限溶剂中 在两根玻璃毛细血管之间。提取的分子被有效地输送到质量中 光谱仪进口和软电喷雾电离。Nano-Desi MSI使 高灵敏度检测组织切片中数百种代谢物、脂质和多肽， 空间分辨率高，无需特殊的样品前处理。此外，在飞行中， 纳米DESI成像实验中组织切片中脂质和代谢物的定量 通过在工作溶剂中掺入已知浓度的适当标准来实现。 该项目将扩展Nano-Desi MSI的这些强大功能，以实现高吞吐量 对HubMAP财团感兴趣的大组织切片进行成像。这将会实现的 使用概念上不同的纳米DESI探针设计的组合 健壮性、易于制造和空间分辨率，以及一套高级机器学习 和压缩传感计算方法。这些发展将适用于 不同类型的人体组织，并将转变为多个定量分子成像 组织切片中的生物分子分类。尽管新的成像平台的功能 将使用非病变组织进行演示，这些开发将广泛适用 与了解健康和疾病相关的科学问题

Novel Platform for Quantitative Subcellular Resolution Imaging of Human Tissues Using Mass Spectrometry Mass spectrometry imaging (MSI) is a powerful technique that enables label-free spatial mapping of different classes of biomolecules in biological systems. Because it does not require any special sample pretreatment, ambient MSI is particularly attractive for high throughput automated imaging applications. The throughput of ambient MSI experiments is typically limited by the inherently slow microprobe-type sampling from surfaces, which is a characteristic shortcoming of many chemical imaging modalities. This project will combine several highly innovative approaches to address challenges associated with the high-throughput high- resolution ambient MSI of lipids and metabolites using nanospray desorption electrospray ionization (nano-DESI). Nano-DESI is an ambient ionization technique, which relies on gentle localized liquid extraction of molecules from tissue sections into a flowing solvent confined between two glass capillaries. The extracted molecules are efficiently delivered to a mass spectrometer inlet and ionized by soft electrospray ionization. Nano-DESI MSI enables detection of hundreds of metabolites, lipids, and peptides in tissue sections with high sensitivity, high spatial resolution, and without special sample pretreatment. Furthermore, on-the-fly quantification of lipids and metabolites in tissue sections during nano-DESI imaging experiments is achieved by doping the working solvent with appropriate standards of known concentration. This project will extend these powerful capabilities of nano-DESI MSI to enable high-throughput imaging of large tissue sections of interest to the HubMAP Consortium. This will be achieved using a combination of a conceptually different nano-DESI probe design optimized for robustness, ease of fabrication, and spatial resolution and a suite of advanced machine learning and compressed sensing computational approaches. These developments will be applicable to different types of human tissues and will transform quantitative molecular imaging of multiple classes of biomolecules in tissue sections. Although the capabilities of the new imaging platform will be demonstrated using non-diseased tissue, these developments will be broadly applicable to scientific problems associated with understanding health and disease