Real-Time Imaging of Mitochondrial Hydrogen Peroxide and pH Fluctuations in Living Cells Using a Fluorescent Nanosensor

2015 ◽  
Vol 87 (7) ◽  
pp. 3678-3684 ◽  
Author(s):  
Limin Yang ◽  
Na Li ◽  
Wei Pan ◽  
Zhengze Yu ◽  
Bo Tang
Keyword(s):  
Hydrogen Peroxide ◽  
Real Time ◽  
Living Cells ◽  
Real Time Imaging ◽  
Ph Fluctuations

A near-infrared fluorescent probe for endogenous hydrogen peroxide real-time imaging in living cells and zebrafish

2019 ◽  
Vol 165 ◽  
pp. 518-523 ◽  
Author(s):  
Xin Huang ◽  
Zhipeng Li ◽  
Zixin Liu ◽  
Chengchu Zeng ◽  
Liming Hu
Keyword(s):  
Hydrogen Peroxide ◽  
Real Time ◽  
Fluorescent Probe ◽  
Near Infrared ◽  
Living Cells ◽  
Real Time Imaging

Real time imaging reveals a peroxisomal reticulum in living cells

2000 ◽  
Vol 113 (20) ◽  
pp. 3663-3671 ◽  
Author(s):  
M. Schrader ◽  
S.J. King ◽  
T.A. Stroh ◽  
T.A. Schroer
Keyword(s):  
Real Time ◽  
Cytoplasmic Dynein ◽  
Living Cells ◽  
Real Time Imaging ◽  
Peroxisomal Targeting ◽  
Microtubule Motor ◽  
Targeting Signal ◽  
Peroxisomal Targeting Signal

We have directly imaged the dynamic behavior of a variety of morphologically different peroxisomal structures in HepG2 and COS-7 cells transfected with a construct encoding GFP bearing the C-terminal peroxisomal targeting signal 1. Real time imaging revealed that moving peroxisomes interacted with each other and were engaged in transient contacts, and at higher magnification, tubular peroxisomes appeared to form a peroxisomal reticulum. Local remodeling of these structures could be observed involving the formation and detachment of tubular processes that interconnected adjacent organelles. Inhibition of cytoplasmic dynein based motility by overexpression of the dynactin subunit, dynamitin (p50), inhibited the movement of peroxisomes in vivo and interfered with the reestablishment of a uniform distribution of peroxisomes after recovery from nocodazole treatment. Isolated peroxisomes moved in vitro along microtubules in the presence of a microtubule motor fraction. Our data reveal that peroxisomal behavior in vivo is significantly more dynamic and interactive than previously thought and suggest a role for the dynein/dynactin motor in peroxisome motility.

scholarly journals Construction of a Nanosensor for Non-Invasive Imaging of Hydrogen Peroxide Levels in Living Cells

Biology ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 430
Author(s):  
Amreen ◽  
Hayssam M. Ali ◽  
Mohammad Ahmad ◽  
Mohamed Z. M. Salem ◽  
Altaf Ahmad
Keyword(s):  
Hydrogen Peroxide ◽  
Real Time ◽  
Mammalian Cells ◽  
Dynamic Range ◽  
Resonance Energy ◽  
Living Cells ◽  
Stress Conditions ◽  
Live Cells ◽  
Cell Physiology ◽  
Broad Dynamic Range

Hydrogen peroxide (H2O2) serves fundamental regulatory functions in metabolism beyond the role as damage signal. During stress conditions, the level of H2O2 increases in the cells and causes oxidative stress, which interferes with normal cell growth in plants and animals. The H2O2 also acts as a central signaling molecule and regulates numerous pathways in living cells. To better understand the generation of H2O2 in environmental responses and its role in cellular signaling, there is a need to study the flux of H2O2 at high spatio–temporal resolution in a real-time fashion. Herein, we developed a genetically encoded Fluorescence Resonance Energy Transfer (FRET)-based nanosensor (FLIP-H2O2) by sandwiching the regulatory domain (RD) of OxyR between two fluorescent moieties, namely ECFP and mVenus. This nanosensor was pH stable, highly selective to H2O2, and showed insensitivity to other oxidants like superoxide anions, nitric oxide, and peroxynitrite. The FLIP-H2O2 demonstrated a broad dynamic range and having a binding affinity (Kd) of 247 µM. Expression of sensor protein in living bacterial, yeast, and mammalian cells showed the localization of the sensor in the cytosol. The flux of H2O2 was measured in these live cells using the FLIP-H2O2 under stress conditions or by externally providing the ligand. Time-dependent FRET-ratio changes were recorded, which correspond to the presence of H2O2. Using this sensor, real-time information of the H2O2 level can be obtained non-invasively. Thus, this nanosensor would help to understand the adverse effect of H2O2 on cell physiology and its role in redox signaling.

Self-Immolative Fluorescent and Raman Probe for Real-Time Imaging and Quantification of γ-Glutamyl Transpeptidase in Living Cells

2019 ◽  
Vol 11 (31) ◽  
pp. 27529-27535 ◽  
Author(s):  
Yang Zhang ◽  
Gengwu Zhang ◽  
Peng Yang ◽  
Basem Moosa ◽  
Niveen M. Khashab
Keyword(s):  
Real Time ◽  
Living Cells ◽  
Real Time Imaging ◽  
Glutamyl Transpeptidase ◽  
Raman Probe

scholarly journals Reactive oxygen species-triggered off-on fluorescence donor for imaging hydrogen sulfide delivery in living cells

2019 ◽  
Vol 10 (33) ◽  
pp. 7690-7694 ◽  
Author(s):  
Yiming Hu ◽  
Xiaoyi Li ◽  
Yu Fang ◽  
Wen Shi ◽  
Xiaohua Li ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Hydrogen Sulfide ◽  
Real Time ◽  
Reactive Oxygen ◽  
Living Cells ◽  
Oxygen Species ◽  
The Real ◽  
Real Time Imaging

A reactive oxygen species-triggered off-on fluorescence H2S donor is develop for the real-time imaging of H2S delivery and the cytoprotection against the hazardous oxidative environment.

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