Year of Award:
Molecular & Cellular Analysis Technologies
FIOLKA, RETO PAUL
Other PI or Project Leader:
UT SOUTHWESTERN MEDICAL CENTER
Fluorescence microscopy is an indispensable tool for the quantitative analysis of themolecular pathways involved in disease. Importantly, heterotypic cell-cell and cell-matrixinteractions within the 3D tumor microenvironment critically influences signaling pathways andhence the tumorigenicity of cancer cells. Therefore, it is important to study cancer cells inphysiologically relevant conditions, which can be realized by using model organisms ororganotypic organoid cultures. However, technological limitations prevent the observation ofcancer cell dissemination and survival throughout an entire organism with sufficientspatiotemporal resolution to evaluate the molecular pathways and oncogenic states of individualcancer cells. Here, we propose to break through this barrier by developing a new opticalmicroscope that is capable of longitudinally imaging an entire organism, but can also providelocal high-resolution imaging. Furthermore, it can also perform 3D photo-manipulation, whichallows temporally and spatially confined perturbation of intracellular signaling, modification ofthe microenvironment, and tagging cells of interest. The most important properties of the newmicroscope are i) multi-scale imaging at cellular and subcellular level, ii) minimal light exposureto allow long observation spans while limiting photo-toxicity, iii) increased optical penetrationdepth via adaptive optics, iv) isotropic spatial resolution in the high-resolution mode and v) 3Dphoto-manipulation using two-photon absorption. Optical modules will be developed that enabletunable light-sheet control, wavefront correction for adaptive optics, 3D scanning for photo-manipulation and variable magnification of the detection path. Thus, for the first time, thisinstrument will allow us to image cancer cell dissemination on an organism scale over extendedtime periods and also monitor and manipulate cell signaling states and morphodynamics withsub-micron, isotropic resolution in metastatic niches.