Diffuse optical spectroscopic imaging (DOSI) is a versatile technology sensitive to changes in tissue composition and hemodynamics and has been used for a wide variety of clinical applications. Specific applications have prompted the development of versions of the DOSI technology to fit specific clinical needs. This work describes the development and characterization of a multi-modal DOSI (MM-DOSI) system that can acquire metabolic, compositional, and pulsatile information at multiple penetration depths in a single hardware platform. Additionally, a 3D tracking system is integrated with MM-DOSI, which enables registration of the acquired data to the physical imaging area. We demonstrate imaging, layered compositional analysis, and metabolism tracking capabilities using a single MM-DOSI system on optical phantoms as well as in vivo human tissue. We characterize system performance with a silicone phantom containing an embedded object. To demonstrate multi-layer sensitivity, we imaged human calf tissue with a 4.8-mm skin-adipose thickness. Human thenar tissue was also measured using a combined broadband DOSI and continuous-wave near-infrared spectroscopy method (∼15 Hz acquisition rate). High-resolution optical property maps of absorption (μa) and reduced scattering (μs ' ) were recovered on the phantom by capturing over 1000 measurement points in under 5 minutes. On human calf tissue, we show two probing depth layers have significantly different (p < 0.001) total-hemo/myoglobin and μs ' composition. On thenar tissue, we calculate tissue arterial oxygen saturation, venous oxygen saturation, and tissue metabolic rate of oxygen consumption during baseline and after release of an arterial occlusion. The MM-DOSI can switch between collection of broadband spectra, high-resolution images, or multi-depth hemodynamics without any hardware reconfiguration. We conclude that MM-DOSI enables acquisition of high resolution, multi-modal data consolidated in a single platform, which can provide a more comprehensive understanding of tissue hemodynamics and composition for a wide range of clinical applications.