Development of a high throughput system for molecular imaging of different cell types in mouse brain tissues

开发用于小鼠脑组织中不同细胞类型的分子成像的高通量系统

• 批准号: 10369883
• 负责人: GAURAV CHOPRA
• 金额: $ 151.14万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2024-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10369883
• 关键词: 3-Dimensional; Address; Anatomy; Atlases; Biological; Brain; Brain imaging; Cells; Censuses; Chemicals; Classification; Coupling; Data; Data Analyses; Data Sources; Data Storage and Retrieval; Development; Disease; Electrospray Ionization; Goals; Graph; Health; Hour; Human; Human BioMolecular Atlas Program; Image; Imaging Device; Imaging technology; Immunofluorescence Immunologic; Immunofluorescence Microscopy; Label; Link; Lipids; Maps; Mass Spectrum Analysis; Measurement; Methods; Microfluidics; Mining; Molecular; Molecular Profiling; Mus; Neuroglia; Neurons; Peptides; Proteins; Proteomics; Research; Resolution; Robotics; Sampling; Scanning; Sensitivity and Specificity; Signal Transduction; Spectrometry, Mass, Electrospray Ionization; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Speed; Structure; System; Techniques; Tissue imaging; Tissues; base; brain cell; brain tissue; cell type; data acquisition; data format; data integration; data mining; data sharing; deep learning; experimental study; high resolution imaging; high throughput technology; image registration; imaging approach; imaging modality; imaging platform; imaging system; improved; innovation; interest; ion mobility; ionization; ionization technique; large scale data; learning strategy; lipidomics; mass spectrometer; member; metabolomics; molecular imaging; nano; novel; tool

Development of a high throughput system for molecular imaging of different cell types in mouse brain tissues Mass spectrometry imaging (MSI) is a powerful tool for developing detailed molecular maps of biological tissues with high specificity and sensitivity. This label-free technique enables simultaneous imaging of multiple classes of molecules including lipids, metabolites, and proteins thereby advancing the understanding of tissue organization and function. In this project, we will advance brain cell census research by developing an innovative platform for the acquisition of a comprehensive spatially-resolved cell-specific atlas of lipids, metabolites, and proteins in mouse brain tissue. The platform will combine MSI with immunofluorescence imaging to place the detailed molecular maps into the spatial cellular context within the brain tissue, which will facilitate image registration to the common coordinate system. Furthermore, we will develop deep learning and data mining approaches to enable automated assignment of molecular signatures to different cell types and efficient data sharing and comparison with other techniques. This research will address the existing bottlenecks in the experimental throughout and molecular coverage of the current MSI technologies along with the limitations of data analysis tools. Furthermore, our approach will compensate for signal suppression during ionization also called ‘matrix effects’, which is particularly severe in imaging of brain tissue. Such matrix effects common to all the MSI modalities including commercial MALDI MSI instruments interfere with the accurate measurement of the spatial localization of molecules. To overcome these challenges, we will advance the capabilities of nanospray desorption electrospray ionization (nano-DESI) - an ambient ionization technique, which efficiently compensates for matrix effects and thereby enables accurate and sensitive imaging of chemical gradients for hundreds of metabolites and lipids in biological tissue sections. Nano-DESI MSI does not require sample pretreatment, has a sub-femtomole sensitivity, and high spatial resolution. The new nano-DESI MSI system will provide a 5-fold increase in the experimental throughput and improve the spatial resolution of protein imaging in whole tissue sections from ~80 µm to ~10 µm. Coupling nano-DESI MSI with a high- resolution ion mobility mass spectrometer will substantially enhance molecular coverage. Meanwhile, co- registration of MSI with immunofluorescence data will be used to generate comprehensive 3D molecular maps of the mouse brain tissue. Collectively, these efforts will establish a robust imaging platform, which will transform our ability to generate detailed molecular maps of different cell types in brain tissues.

小鼠脑内不同细胞类型高通量分子成像系统的研制 组织 摘要质谱成像(MSI)是绘制生物分子图谱的有力工具。 具有高度特异性和敏感性的组织。这种无标记技术能够同时成像 包括脂质、代谢物和蛋白质在内的多种分子，从而促进 了解组织的组织和功能。在这个项目中，我们将推进脑细胞普查 通过开发创新平台进行研究，获取空间分辨率全面的 小鼠脑组织中脂质、代谢物和蛋白质的细胞特异性图谱。该平台将结合MSI 通过免疫荧光成像将详细的分子图谱置于空间细胞环境中 在脑组织内，这将有助于图像配准到公共坐标系。 此外，我们将开发深度学习和数据挖掘方法，以实现自动作业 不同细胞类型的分子签名以及高效的数据共享和与其他 技巧。这项研究将解决整个实验过程中存在的瓶颈和 当前MSI技术的分子覆盖面以及数据分析工具的局限性。 此外，我们的方法将补偿电离过程中的信号抑制，也称为矩阵 影响‘，这在脑组织成像中尤其严重。这种矩阵效应对所有MSI都是通用的 包括商用MALDI MSI仪器在内的医疗设备会干扰准确测量 分子的空间局部化。为了克服这些挑战，我们将推进 纳米解吸电喷雾电离(Nano-DESI)-一种环境电离技术，它 有效地补偿基质效应，从而实现准确和灵敏的化学成像 生物组织切片中数百种代谢物和脂质的梯度。Nano-Desi MSI不需要 样品前处理，具有亚毫微克分子灵敏度，且空间分辨率高。新的纳米DISI MSI 系统将使实验吞吐量提高5倍，并提高空间分辨率 在从~80微米到~10微米的整个组织切片中进行蛋白质成像。 分辨率离子迁移率质谱仪将大大提高分子覆盖率。同时，联合- MSI与免疫荧光数据的注册将用于生成全面的3D分子 小鼠脑组织的地图。总体而言，这些努力将建立一个强大的成像平台，这将 改变我们生成脑组织中不同细胞类型的详细分子图谱的能力。