A multimodal workflow for mass spectrometry imaging was developed that combines MALDI imaging with protein identification and quantification by liquid chromatography tandem mass spectrometry (LC-MS/MS). Thin tissue sections were analyzed by MALDI imaging, and the regions of interest (ROI) were identified using a smoothing and edge detection procedure. A midinfrared laser at 3-μm wavelength was used to remove the ROI from the brain tissue section after MALDI mass spectrometry imaging (MALDI MSI). The captured material was processed using a single-pot solid-phase-enhanced sample preparation (SP3) method and analyzed by LC-MS/MS using ion mobility (IM) enhanced data independent acquisition (DIA) to identify and quantify proteins; more than 600 proteins were identified. Using a modified database that included isoform and the post-translational modifications chain, loss of the initial methionine, and acetylation, 14 MALDI MSI peaks were identified. Comparison of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of the identified proteins was achieved through an evolutionary relationships classification system.

中文翻译：

---

MALDI成像指导激光消融组织微量采样，用于独立于数据的采集蛋白质组学。

开发了质谱成像的多模式工作流程，该流程将MALDI成像与蛋白质鉴定和液相色谱串联质谱（LC-MS / MS）定量相结合。通过MALDI成像分析薄组织切片，并使用平滑和边缘检测程序识别感兴趣区域（ROI）。在MALDI质谱成像（MALDI MSI）之后，使用波长为3μm的中红外激光从大脑组织切片中去除ROI。使用单锅固相增强样品制备（SP3）方法处理捕获的材料，并使用离子迁移率（IM）增强数据独立采集（DIA）通过LC-MS / MS进行分析，以鉴定和定量蛋白质。鉴定出600多种蛋白质。使用包含同工型和翻译后修饰链，初始甲硫氨酸丢失和乙酰化的修饰数据库，鉴定出14个MALDI MSI峰。通过进化关系分类系统，比较了所鉴定蛋白质的《京都基因与基因组百科全书》（KEGG）途径。