1. Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana‐Champaign, Urbana, IL, USA;2. Department of Electrical and Computer Engineering, University of Illinois at Urbana‐Champaign, Urbana, IL, USA;3. Department of Bioengineering, University of Illinois at Urbana‐Champaign, Urbana, IL, USA;4. Department of Internal Medicine, University of Illinois at Urbana‐Champaign, Urbana, IL, USA
Abstract:
Cell death plays a critical role in health and homeostasis as well as in the pathogenesis and treatment of a broad spectrum of diseases and can be broadly divided into two main categories: apoptosis, or programmed cell death, and necrosis, or acute cell death. While these processes have been characterized extensively in vitro, label‐free detection of apoptosis and necrosis at the cellular level in vivo has yet to be shown. In this study, for the first time, fluorescence lifetime imaging microscopy (FLIM) of intracellular reduced nicotinamide adenine dinucleotide (NADH) was utilized to assess the metabolic response of in vivo mouse epidermal keratinocytes following induction of apoptosis and necrosis. Results show significantly elevated levels of both the mean lifetime of NADH and the intracellular ratio of protein bound‐to‐free NADH in the apoptotic compared to the necrotic tissue. In addition, the longitudinal profiles of these two cell death processes show remarkable differences. By identifying and extracting these temporal metabolic signatures, apoptosis in single cells can be studied in native tissue environments within the living organism.
