This TECH core of the U54 Center for Multiparametric Imaging of Tumor Immune Microenvironments (C-MITIE) will develop an integrated toolkit of advanced imaging and data analysis to power quantitative, mechanistic investigations of immune-microenvironment dynamics in poor prognosis solid tumors. There is great need for improved imaging methods that can advance understanding of the physical and molecular mechanisms governing immune infiltration, distribution, and function in native tumor microenvironments. We propose a number of multiparametric imaging and computational methods for the two research test beds that seek to define the physical and molecular barriers to effective anti-tumor immunity and immunotherapies. A major theme of the TECH approach is to use label-free imaging approaches that can characterize and quantitate the interactions between immune cells and the tumor microenvironment. These label free methods are largely built on the platform method of multiphoton microscopy and can be used on intact cell and tissue models with minimal perturbation. T-cell identity and activation will be tracked by metabolic profiling using new fluorescence lifetime (FLIM) and hyperdimensional imaging (HDIM) approaches. These metabolically sensitive methods will be complemented by Full-Field Optical Coherence Tomography (FFOCT) to reveal new insight into metabolically relevant architecture. FLIM based FRET can be used to yield new insights into signaling molecular interactions relevant to immune-microenvironment dynamics The collagen rich extracellular matrix (ECM) will be queried with Second Harmonic Generation (SHG) imaging for collagen fiber topology measurement and collagen cross- linking measurements with Enhanced Backscattering Spectroscopy (EBS). Multiphoton Excitation (MPE) photochemistry fabrication can be used to create in vitro cell ready models of collagen fiber organization that are directly based on human data blueprints. Advanced computational analysis methods including algorithmic and machine learning approaches will be used to examine all multiparametric signals and make correlation between immune and microenvironment interactions. All imaging and computational methods will be shared not only widely within the UW and UMN research teams but importantly with the general cancer imaging community using established hardware and open source software dissemination protocols.