The analysis of different central nervous system regions is interesting and necessary as each one is involved in specific physiological processes and pathologies. On that matter, differences in the chemical composition between the same brain regions in different mice strains have been reported. In this sense, the development of a simple method for the identification of these regions depending on their chemical composition becomes relevant. Raman microspectroscopy, a non-destructive analytical chemical approach for biological samples, is a widely used method for qualitative, quantitative, and structural analysis in biochemical research. Ten brain structures in three different mice strains (Triple transgenic for Alzheimer Disease, 3xTgAD; Cluster 57 black 6, C57BL/6; and the Swiss strain, CD1) were analyzed, and variations among samples from several brain regions were found. Particularly, the pattern of signals from the hippocampus, the prefrontal and temporal cortices, the basal forebrain, the striatum, the cerebellum, and the hypothalamus was discernable. Interestingly, notable signals regarding non-peptide small neurotransmitters were observed, including those related to acetylcholine. These bands were present in the Raman spectra of the basal forebrain and prefrontal cortex in the three mice strains, consistent with the relative abundance of this neurotransmitter in those regions. However, signals with lower intensities appeared in the basal forebrain of C57BL/6 in comparison with the same tissue of the other two strains. In addition, the Raman intensity of bands assigned to catecholamines in the striatum was lower in the 3xTgAD than those from both CD1 and C57BL/6 mice strains. This approach, as well as the reported differences, has potential application for designing analysis on specific murine models of brain diseases.