Ju Hwan Song (Korea Basic Science Institute)

Ju Yeon Lee (Korea Basic Science Institute)

Ju Hwan Song (Korea Basic Science Institute)

Hyeon-gu Yeo (Korea Research Institute of Bioscience & Biotechnology)

Kyoung Heon Kim (Korea University)

Eun Hee Han (Korea Basic Science Institute)

Youngjeon Lee (Korea Research Institute of Bioscience & Biotechnology)

Ju Yeon Lee (Korea Basic Science Institute)

Spatial proteomics (SP) has emerged as a powerful approach for investigating mechanisms of health and disease, providing unprecedented insights into the spatial organization of proteins within cells and tissues. As a rapidly advancing field, SP has garnered significant attention. Ultrasensitive mass spectrometry provide the enabled multiscale spatial proteomics, integrating complementary imaging techniques with exploratory mass spectrometry approaches to achieve highly detailed spatial protein analysis.

 In this study, to ensure compatibility with low-input samples, we confirmed the reproducibility and data reliability of an one-pot digestion workflow using nanogram-scale proteome sample preparation and  LC-MS/MS systestm consisting of an Ultimate 3000RSLnano System and Orbitrap Fusion Lumos Tribrid mass spectrometer. And then, this protocol was appiled to monkey brain tissue spatially sectioned via laser microdissection (LMD) and the resulting proteome smaples were analyzed using LC-MS/MS systestm  achieving high sensitivity and consistent quantification from minimal sample input.