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Updated: May 19, 2026
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Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells
Published on: March 16, 2017
A small-molecule two-photon probe for nitric oxide in living tissues
Eun Won Seo1, Ji Hee Han, Cheol Ho Heo
1Division of Energy Systems Research, Ajou University, Suwon, 443-749 Korea.
Chemistry (Weinheim an Der Bergstrasse, Germany)
|August 22, 2012
Summary
Researchers developed a novel small-molecule probe (ANO1) for detecting nitric oxide (NO) using two-photon microscopy (TPM). This probe offers rapid, specific NO detection with significant fluorescence enhancement, enabling real-time imaging in living tissues.
Area of Science:
- Biomedical Imaging
- Chemical Biology
- Molecular Probes
Background:
- Two-photon microscopy (TPM) is crucial for deep tissue molecular imaging in biology and medicine.
- Development of specific probes is essential for maximizing TPM's utility.
- Nitric oxide (NO) is a vital signaling molecule requiring precise detection methods.
Purpose of the Study:
- To develop and characterize a novel small-molecule two-photon probe for nitric oxide (NO).
- To evaluate the probe's performance in terms of response speed, specificity, and fluorescence enhancement.
- To demonstrate the probe's capability for real-time NO monitoring in living biological systems using TPM.
Main Methods:
- Synthesis and characterization of the small-molecule two-photon probe (ANO1).
- In vitro assessment of NO response, fluorescence enhancement, and TP-action cross-section.
- Cell and tissue loading studies.
- Real-time in vivo imaging of NO in living tissues using TPM at depths of 100-180 μm.
Main Results:
- The ANO1 probe exhibits a rapid and specific response to nitric oxide.
- A significant 68-fold fluorescence enhancement was observed upon reaction with NO.
- The probe achieved a maximum two-photon action cross-section of 170 GM.
- ANO1 was successfully loaded into cells and tissues, allowing for real-time NO monitoring.
- In vivo imaging demonstrated NO detection at depths of 100-180 μm for over 1200 seconds with minimal interference.
Conclusions:
- ANO1 is a highly effective small-molecule two-photon probe for sensitive and specific detection of nitric oxide.
- The probe's properties facilitate real-time monitoring of NO dynamics in living tissues using TPM.
- This development provides a valuable tool for advancing biological and medical research involving nitric oxide signaling.
