Related Experiment Video
Updated: Jun 30, 2026
Time-Lapse Video Microscopy for Assessment of EYFP-Parkin Aggregation as a Marker for Cellular Mitophagy
Published on: May 4, 2016
Unveiling the hypoxia-induced mitophagy process through two-channel real-time imaging of NTR and viscosity under the
Lei Wen1, Mengqi Shao2, Yinhui Li1
1Key Laboratory for Green Organic Synthesis and Application of Hunan Province, Key Laboratory of Environmentally Friendly Chemistry and Application of Ministry of Education, College of Chemistry, Xiangtan University, Xiangtan, 411105, China.
Abstract:
Mitophagy is an essential physiological process that eliminates damaged mitochondria via lysosomes. It is reported that hypoxia, inflammatory stimuli or other stress conditions could lead to mitochondrial damage and mitochondrial dysfunction, which induces the process of mitophagy. Herein, we report a novel fluorescent probe PC-NTR for imaging hypoxia-induced mitophagy by monitoring the change of nitroreductase and viscosity simultaneously. To our delight, PC-NTR could respond simultaneously to nitroreductase and viscosity at different fluorescence channels with no mutual interference under the same excitation wavelength. The fluorescence emission around 535 nm was enhanced dramatically after addition of nitroreductase while the fluorescence emission around 635 nm heightened as the viscosity increased. The probe would be able to selectively targeting of mitochondria in cells because of the positively charged pyridine salt structure of PC-NTR. The probe was successfully applied to assess the different levels of hypoxia and real-time imaging of mitochondrial autophagy in live cells. More importantly, using dual channel imaging, PC-NTR could be used to distinguish cancer cells from normal cells and was successfully applied to imaging experiments in HeLa-derived tumor-bearing nude mice. Therefore, PC-NTR would be an important molecular tool for hypoxia imaging and detecting solid tumors in vivo.
Insights
A new fluorescent probe, PC-NTR, images hypoxia-induced mitophagy by monitoring nitroreductase and viscosity. This tool aids in distinguishing cancer cells from normal cells and in vivo tumor detection.
Area of Science:
- Biomedical Imaging
- Molecular Probes
- Cellular Biology
Background:
- Mitophagy is crucial for removing damaged mitochondria, often triggered by cellular stress like hypoxia.
- Monitoring mitophagy is vital for understanding cellular dysfunction and disease progression.
Purpose of the Study:
- To develop a novel fluorescent probe (PC-NTR) for simultaneous imaging of hypoxia-induced mitophagy.
- To enable real-time monitoring of nitroreductase activity and viscosity changes within mitochondria.
Main Methods:
- Design and synthesis of PC-NTR, a mitochondria-targeting fluorescent probe.
- Utilizing dual-channel fluorescence detection to monitor nitroreductase and viscosity.
- Application of PC-NTR in live cell imaging and in vivo studies in tumor-bearing mice.
Main Results:
- PC-NTR exhibits distinct fluorescence responses to nitroreductase (535 nm) and viscosity (635 nm) without cross-interference.
- The probe successfully images hypoxia-induced mitophagy in live cells.
- PC-NTR differentiates cancer cells from normal cells and detects tumors in vivo.
Conclusions:
- PC-NTR is a versatile dual-channel fluorescent probe for imaging hypoxia and mitophagy.
- This probe serves as a valuable tool for cancer cell identification and in vivo tumor imaging.
