Profiling cells in suspension offers a high-throughput approach for analyzing RNA and proteins at single-cell resolution. However, upon tissue dissociation, all spatial context is lost. Spatial information can offer insights into cell-cell interactions and the nature of the cellular niche and its microenvironmental cues. We are actively implementing high-dimensional spatial profiling methodologies and developing analysis pipelines for spatial profiling of the brain and tumor immune landscape. Additionally, we are complementing this with 2-photon intravital microscopy, to study brain macrophage dynamics in steady-state and disease.