The intravital imaging method described here utilizes collagen second harmonic generation and endogenous fluorescence from the metabolic co-factor NAD(P)H to non-invasively segment an unlabeled tumor microenvironment into tumor, stromal, and vascular compartments for in-depth analysis of 4D intravital images.
Burkel, B. M., Inman, D. R., Virumbrales-Muñoz, M., Hoffmann, E. J., Ponik, S. M. A Label-Free Segmentation Approach for Intravital Imaging of Mammary Tumor Microenvironment. J. Vis. Exp. (183), e63413, doi:10.3791/63413 (2022).