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Specific Labeling of Mitochondrial Nucleoids for Time-lapse Structured Illumination Microscopy
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A non-toxic fluorogenic dye for mitochondria labeling.

Junyan Han1, Myung Shin Han, Ching-Hsuan Tung

  • 1Department of Translational Imaging, The Methodist Hospital Research Institute, Weill Cornell Medical College, Houston, TX, USA.

Biochimica Et Biophysica Acta
|July 16, 2013
PubMed
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A new non-toxic dye, AcQCy7, allows for long-term, bright fluorescence imaging of mitochondria in live cells. This mitochondria-specific probe enables clear real-time monitoring of mitochondrial dynamics without cytotoxicity, aiding disease research.

Area of Science:

  • Cell Biology
  • Mitochondrial Research
  • Fluorescence Imaging

Background:

  • Mitochondria are vital organelles regulating cellular energy, reactive oxygen species, and apoptosis.
  • Understanding mitochondrial morphology is crucial for elucidating disease pathogenesis and functional regulation.
  • Live-cell imaging of mitochondria requires specific, non-toxic probes for temporal and spatial monitoring.

Purpose of the Study:

  • To develop and validate a novel, non-cytotoxic, mitochondria-specific fluorogenic dye.
  • To enable long-term, high-resolution fluorescence imaging of mitochondrial dynamics in live cells.
  • To assess the probe's utility in monitoring mitochondrial changes, such as fission during apoptosis.

Main Methods:

  • Synthesis of a novel fluorogenic compound, AcQCy7, designed for mitochondria-specific targeting.
Keywords:
ApoptosisCyanine dyeCytotoxicityFluorogenic dyeMitochondria

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  • Evaluation of AcQCy7's fluorescence properties, intracellular activation, and retention within mitochondria.
  • Assessment of AcQCy7's cytotoxicity compared to a standard mitochondria probe (Mitochondria Tracker Green) over a 2-day culture period.
  • Real-time monitoring of apoptosis-induced mitochondrial fission using AcQCy7.

    • Main Results:

    • AcQCy7 is activated intracellularly via hydrolysis, producing a bright, mitochondria-localized fluorescent signal.
    • The probe exhibits excellent retention in mitochondria, facilitating prolonged imaging without cell washing.
    • AcQCy7 demonstrated no significant cytotoxicity over 2 days, unlike Mitochondria Tracker Green, which caused substantial cell death.
    • The dye clearly visualized mitochondria fission events associated with apoptosis in real-time.

    Conclusions:

    • AcQCy7 is a validated, simple add-and-read mitochondria-specific dye effective across various cell models.
    • The probe provides bright, specific mitochondrial fluorescence lasting several days with negligible toxicity.
    • AcQCy7 is a non-toxic agent suitable for long-term mitochondria imaging and dynamic studies.