The tumor microenvironment plays a critical role in influencing the behavior and characteristics of tumor cells, particularly in solid tumors such as glioblastoma (GBM). A key study by Kumar highlights the importance of spatial positioning of tumor cells relative to blood vessels, which significantly affects their exposure to microenvironmental signals. The research introduces a novel methodology for isolating tumor cells based on their distance from blood vessels, utilizing a fluorescent dye that differentially labels cells depending on their proximity to vasculature. This technique allows for the sorting of stroma-free tumor cells from various microenvironments, facilitating subsequent multi-OMICs and functional analyses (ref: Kumar doi.org/10.21769/BioProtoc.3628/). The findings underscore the heterogeneity within tumors, as cells located in different microenvironments exhibit distinct phenotypic traits and responses to therapies, which could have profound implications for personalized treatment strategies. Moreover, the study emphasizes the necessity of understanding the tumor microenvironment's influence on cell behavior, as it can lead to the identification of novel therapeutic targets and biomarkers. The methodology presented not only enhances our ability to study tumor cell heterogeneity but also provides a framework for future research aimed at unraveling the complex interactions between tumor cells and their microenvironment. By isolating cells based on their spatial context, researchers can gain insights into how these interactions contribute to tumor progression and treatment resistance, ultimately paving the way for more effective cancer therapies.