The present invention provides a computer system and user interface that allows a user to readily view and analyze two-dimensional and three-dimensional in vivo images and imaging data. The user interface is well-suited for one or more of the following actions pertinent to in vivo light imaging: investigation and control of three-dimensional imaging data and reconstruction algorithms; control of topographic reconstruction algorithms; tomographic spectral imaging and analysis; and comparison of two-dimensional or three-dimensional imaging data obtained at different times.