Two-photon dual imaging platform for in vivo monitoring cellular oxidative stress in liver injury

Abstract

Oxidative stress reflects an imbalance between reactive oxygen species (ROS) and antioxidants, which has been reported as an early unifying event in the development and progression of various diseases and as a direct and mechanistic indicator of treatment response. However, highly reactive and short-lived nature of ROS and antioxidant limited conventional detection agents, which are influenced by many interfering factors. Here, we present a two-photon sensing platform for in vivo dual imaging of oxidative stress at the single cell-level resolution. This sensing platform consists of three probes, which combine the turn-on fluorescent transition-metal complex with different specific responsive groups for glutathione (GSH), hydrogen peroxide (H2O2) and hypochlorous acid (HOCl). By combining fluorescence intensity imaging and fluorescence lifetime imaging, these probes totally remove any possibility of crosstalk from in vivo environmental or instrumental factors, and enable accurate localization and measurement of the changes in ROS and GSH within the liver. This precedes changes in conventional biochemical and histological assessments in two distinct experimental murine models of liver injury. The ability to monitor real-time cellular oxidative stress with dual-modality imaging has significant implications for high-accurate, spatially configured and quantitative assessment of metabolic status and drug response.

Metabolic imaging and secondary ion mass spectrometry to define the structure and function of liver with acute and chronic pathology

“…31 Another work demonstrated the study of cellular oxidative stress in liver injury using multiphoton microscopy. 9 Fluorescence intensity imaging, FLIM, and sensing probes for glutathione and reactive oxygen species were combined to detect oxidative stress in the livers of living mice. This approach was proposed as a tool for monitoring cellular oxidative stress, to predict drug responses in the most common types of liver injury.…”

“…This approach was proposed as a tool for monitoring cellular oxidative stress, to predict drug responses in the most common types of liver injury. 9 Here, we have presented studies of the liver metabolism, lipid composition, and fibrous structures during the progression of acute and chronic pathologies using fluorescence intensity imaging, FLIM, and TOF-SIMS without any probes or labeling. We hope that our data will help expand understanding of the processes that occur during the development of liver pathology, as well as complement recent works on the study of the structure and function of the liver using multiphoton microscopy.…”

“…Moreover, it enables identification of different endogenous and exogenous fluorophores that have overlapping or poorly defined spectra but that have different fluorescence lifetimes. 9,10 TOF-SIMS is a sensitive, label-free technique used for both routine composition analysis and for chemical mapping of cells and tissues. [11][12][13][14] By comparing the spectra of control and affected tissues, differences in the chemical composition can be revealed.…”

Two-photon microscopy imaging of oxidative stress in human living erythrocytes

“…Moreover, multiphoton imaging provided a direct observation of amyloid-β-related oxidative stress in senile plaques in Alzheimer's models of living mice [51]. A very important field is the development of molecular probes for two-photon fluorescence intensity imaging of biomarkers of oxidative stress, such as fluorescent transition-metal complexes with different specific responsive groups for detection of glutathione (GSH), hydrogen peroxide (H 2 O 2 ) and hypochlorous acid (HOCl) [52], two-photon pro-fluorescent nitroxides for the detection and imaging of ROS [53], as well as for detection of oxidative stress-induced bright autofluorescent appearance inside and around retinal pigment epithelial cells [49]. The non-invasive and sensitive label free methods of two-photon microscopy have beneficially been used also for the autofluorescent studies of the intrinsic signals coming from endogenous fluorophores, such as autofluorescent metabolic products collagen, retinol, retinoic acid, porphyrin, autofluorescent metabolic coenzymes, such as flavin adenine dinucleotide (FAD) and nicotinamide adenine dinucleotide (NADH) [54,55], autofluorescence of oxidized lipids [56].…”

Rapid Response Fluorescence Probe Enabled In Vivo Diagnosis and Assessing Treatment Response of Hypochlorous Acid‐Mediated Rheumatoid Arthritis

“…Of these methods, fluorescent probes have been proven to be an indispensable tool for real‐time visualization and analysis of the localization and dynamics metabolism of HOCl in biological systems, owing to their versatile advantages such as high sensitivity, simplicity for implementation, real‐time detection, and good compatibility for biological samples 12. For many years, we have also focused on the development of novel bioanalytical methods for HOCl detection in biological systems, especially the HOCl generated in inflammatory diseases, such as drug‐induced liver injury and ischemia–reperfusion (I/R) injury 13. As part of our ongoing research, we recently focused on engineering rapid and effective HOCl responsive probes for unveiling the detailed roles of HOCl in RA diagnosis and further evaluating the treatment response of RA by antiarthritic drug.…”