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Related Experiment Videos

Fluorescent Indicators for Imaging Nitric Oxide Production.

Kojima1, Urano, Kikuchi

  • 1Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan).

Angewandte Chemie (International Ed. in English)
|November 11, 1999
PubMed
Summary

This study introduces DAF-FM DA, a novel indicator for detecting nitric oxide (NO) production within cells. Fluorescence microscopy visualizes NO generation by NO synthase (NOS), enabling precise localization of this crucial signaling molecule.

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Area of Science:

  • Cellular biology
  • Biochemistry
  • Molecular imaging

Background:

  • Nitric oxide (NO) is a critical signaling molecule in various physiological processes.
  • Accurate detection and localization of intracellular NO production are essential for understanding its biological roles.
  • Existing methods for NO detection may have limitations in sensitivity or specificity.

Purpose of the Study:

  • To introduce and validate the use of DAF-FM DA as a sensitive indicator for intracellular nitric oxide (NO) production.
  • To demonstrate the capability of fluorescence microscopy in visualizing NO generation and localization within cells.
  • To provide a tool for studying the activity of NO synthase (NOS) in cellular contexts.

Main Methods:

  • Utilizing the membrane-permeating indicator DAF-FM DA.

Related Experiment Videos

  • Intracellular esterase-mediated conversion of DAF-FM DA to the water-soluble dye DAF-FM.
  • NO trapping by DAF-FM to form a fluorescent triazole compound.
  • Monitoring NO production and localization using fluorescence microscopy.

    • Main Results:

    • DAF-FM DA is efficiently converted to DAF-FM intracellularly.
    • The resulting triazole compound exhibits high fluorescence, allowing for sensitive detection of NO.
    • Fluorescence microscopy successfully identified the intracellular production and precise location of NO.
    • The method allows for direct visualization of NO generated by NO synthase (NOS).

    Conclusions:

    • DAF-FM DA is a valuable tool for the direct identification and localization of intracellular nitric oxide (NO) production.
    • This method facilitates the study of NO synthase (NOS) activity and NO signaling pathways.
    • Fluorescence microscopy combined with DAF-FM DA offers a robust approach for NO research.