Self-Referenced Probe Enables Quantified Monitoring Mitochondrial Membrane Potential by Colocalization Coefficients.

Area of Science:

• Cell Biology
• Biochemistry
• Molecular Imaging

Background:

• Mitochondria are vital organelles with dynamic physiological states.
• Mitochondrial membrane potential (MMP) is crucial for cellular health and viability.
• Accurate, real-time monitoring of MMP dynamics is challenging but essential.

Purpose of the Study:

• To design and synthesize a novel fluorescent probe for quantitative MMP tracking.
• To develop a method for synchronous and in situ monitoring of MMP.
• To provide a new strategy for understanding mitochondria-related biological processes.

Main Methods:

• Synthesis of a novel pH-sensitive, two-color fluorescent probe (CBTH) utilizing spirocyclization.
• Utilizing electrostatic interactions for dual-color mitochondrial targeting.
• Quantifying MMP changes via fluorescence signal migration and colocalization coefficients.

Main Results:

• CBTH successfully targets mitochondria in cells with normal MMP.
• MMP reduction causes a distinct red fluorescence shift to the nucleus, while green fluorescence remains in mitochondria.
• The probe exhibits reversible detection and a direct proportional relationship with MMP in cellular experiments.

Conclusions:

• CBTH offers a novel strategy for quantitative and dynamic MMP detection.
• The probe's response mechanism provides a reliable method for assessing cellular health.
• This work facilitates the development of advanced probes for mitochondrial research.