Updated project metadata. Quantitative protein mapping on whole-tissue levels provides important insights into the spatially-organized regulatory processes/networks related to diseases and therapy, but remains a tremendous challenge. We describe a micro-scaffold assisted spatial proteomics(MASP) method, based on precise tissue spatial-compartmentalization using a 3D-printed micro-scaffold, capable of mapping thousands of proteins across a whole-tissue slice with excellent quantitative quality. The mapping accuracy was validated and applied in mapping >5,000 cerebral proteins in mouse brain. Under stringent cutoffs, 5019 unique proteins were mapped(N=208 micro-specimens) and 4577 proteins were mapped in all regions.