We describe a novel digital light processing, DLP hyperspectral imaging system for visualizing chemical composition of in vivo tissues during surgical procedures non-invasively and at near video rate. The novelty of the DLP hyperspectral imaging system resides in (1) its ability to conform light to rapidly sweep through a series of preprogrammed spectral illuminations as simple as a set of contiguous bandpasses to any number of complex spectra, and (2) processing the reflected spectroscopic image data using unique supervised and unsupervised chemometric methods that color encode molecular content of tissue at each image detector pixel providing an optical biopsy. Spectral illumination of tissue is accomplished utilizing a DLP® based spectral illuminator incorporating a series of bandpass spectra and measuring the reflectance image with a CCD array detector. Wavelength dependent images are post processed with a multivariate least squares analysis method using known reference spectra of oxy- and deoxy-hemoglobin. Alternatively, illuminating with complex reference spectra reduces the number of spectral images required for generating chemically relevant images color encoded for relative percentage of oxyhemoglobin are collected and displayed in real time near-video rate, (3 to 4) frames per second (fps). As a proof of principle application, a kidney of an anesthetized pig was imaged before and after renal vasculature occlusion showing the clamped kidney to be 61% of the undamped kidney percentage of oxyhemoglobin. Using the "3-Shot" spectral illumination method and gathering data at (3 to 4) fps shows a non-linear exponential de-oxygenation of hemoglobin reaching steady state within 30 seconds post occlusion.