Quantitative Single Cell Phenotypic Imaging of Multicellular Systems

Biological systems are highly complex due to the intricate spatial organizations of different types of cells and the heterogeneous nature of living cells. Characterizing the phenotypic behavior of single cells within their native microenvironment is crucial in revealing the pathophysiology of tissue. Current tools for studying multicellular systems are often limited to destructive biochemical analysis, characterization of dissociated cells or molecule-specific imaging with fluorescent labels. These limitations prohibit assessment of key phenotypic properties such as cell size and growth, metabolic status, microenvironmental change, and cell interactions. We developed a multimodal multiphoton optical imaging system to address this challenge. It incorporates two-photon fluorescence, transient absorption microscopy, and stimulated Raman scattering microscopy on the same platform, which enables comprehensive imaging of cell shape, size, density, growth, metabolism, type, and interaction. The platform is applicable to not only cell cultures in 2D and 3D, but also living animals. We aim to employ this platform to study a wide range of biological questions including tumor development, drug transport, and neuro-vascular coupling.