Real-Time, Noninvasive Evaluation of Mitochondrial Function Using Resonance Raman Spectroscopy

Although mitochondria are central to pathogenesis and disease progression, mechanistic insight into mitochondrial health dynamics remains costly and inaccessible. To address this gap, this study employs Resonance Raman Spectroscopy (RRS) to assess mitochondrial function, using a portable system that delivers real-time, noninvasive, and quantitative measurements of mitochondrial cytochrome redox states in rat livers. In this protocol, we demonstrate the use of this technology, including setup, data acquisition, and data processing. This study presents a proof-of-concept experiment that highlights RRS's ability to measure real-time changes in mitochondrial redox state- and, by extension, mitochondrial function. Briefly, the RRS device was connected to a laser pump as well as a data acquisition computer and placed 1 cm away from the rat liver.  Acquisition parameters were selected in accordance with the rat liver protocol; redox states were measured in oxygenated and ischemic conditions utilizing an oxygen stress test. Changes in mitochondrial redox states were tracked throughout the oxygen stress test.