Targeted approaches have not been successful in Glioblastoma (GBM) due to extreme heterogeneity and despite aggressive chemotherapy, radiation, and surgery, only 5.6% of patients survive beyond 5 years post-diagnosis. Mosaic amplification of oncogenes suggests that multiple genetically distinct clones are present in each tumor. However, little is known about how different subpopulations of GBM cells interact with each other or with the surrounding tumor microenvironment (TME). To address this, spatial proteomics coupled with single-cell spatial maps of fluorescence in situ hybridization (FISH) for key oncogenes frequently amplified in GBM were used to analyze 3-4 areas from 17 formalin-fixed paraffin-embedded (FFPE) GBM cases, 96 sub-regions of interest and a total of 35,843 single nuclei.