Visualizing the presence and distribution of a number of particular molecular markers inside a tumor can reveal the composition of its microenvironment, inform prognosis, stratify sufferers, and information therapy. Raman imaging with a number of molecularly-targeted floor enhanced Raman scattering (SERS) nanoprobes might assist examine rising most cancers therapies preclinically or allow personalised therapy evaluation. Right here, we report a complete technique for multiplexed imaging utilizing SERS nanoprobes and machine studying (ML) to watch the early results of immune checkpoint blockade (ICB) in tumor-bearing mice. We used antibody-functionalized SERS nanoprobes to visualise 7+1 immunotherapy-related targets concurrently. The multiplexed pictures had been spectrally resolved after which spatially segmented into superpixels primarily based on the unmixed alerts. The superpixels had been used to coach ML fashions, resulting in the profitable classification of mice into handled and untreated teams, and figuring out tumor areas with variable responses to therapy. This technique could assist predict therapy efficacy in tumors and establish areas of tumor variability and remedy resistance.