Imaging of targeted fluorescent probes offers significant advantages for investigating disease and tissue function in animal models in vivo. Conversely, macroscopic tomographic imaging is challenging because of the high scatter of light in biological tissue and the ill-posed nature of the reconstruction mathematics. In this work, we use the earliest-transmitted photons through Lewis Lung Carcinoma bearing mice, thereby dramatically reducing the effect of tissue scattering. By using a fluorescent probe sensitive to cysteine proteases, the method yielded outstanding imaging performance compared with conventional approaches. Accurate visualization of biochemical abnormalities was achieved, not only in the primary tumor, but also in the surrounding tissue related to cancer progression and inflammatory response at the organ level. These findings were confirmed histologically and with ex vivo fluorescence microscopy. The imaging fidelity demonstrated underscores a method that can use a wide range of fluorescent probes to accurately visualize cellular-and molecular-level events in whole animals in vivo. (5)] or identifying proteins and molecular pathways of development, function, and disease in vivo. Further enhanced by sensing of spectrally separated fluorescent molecules, fluorescence microscopy currently offers unparalleled insights in the study of cellular and protein interactions. In principle, fluorescence technology could be similarly used to study normal and diseased tissues at the whole-organ and live-animal level. However, there remain significant challenges in imaging beyond a few hundred micrometers in depth because of the high degree of light scatter in tissues, which arises from photon interactions with cellular membranes and organelles. Currently, fluorescence macroscopic imaging is largely performed by illuminating tissue at the fluorochrome's excitation wavelength and detecting the emission from within the animal's body by using appropriate spectral filters. Photographic and planar-fluorescence-imaging implementations of this approach offer only 2-dimensional views and largely qualitative data, because they do not consider the nonlinear relation of photon strength to lesion depth and tissue optical properties. In response, tomographic approaches have been developed to overcome the limitations of these methods (6-8). Optical tomography utilizes multiprojection measurements and physical models of photon propagation to quantitatively retrieve fluorochrome biodistribution in tissues. However, the physical limits of imaging performance of current tomographic implementations ultimately depend on the tissue optical properties-primarily scattering-which leads to a highly ill-posed (i.e., inaccurate) inverse problem that reduces the spatial resolution and yields images that are highly dependent on the particulars of the reconstruction algorithm used.To develop a next-generation optical tomographic method that can selectively reduce the effects of tissue scattering to improve imaging accuracy, we developed a sy...