Simultaneous Dynamic Imaging of pH and Temperature during Cellular Metabolism with a Dual-Channel Fluorescent Probe

Area of Science:

• Biomedical Engineering
• Cellular Physiology
• Fluorescent Probes

Background:

• Simultaneous dynamic monitoring of cellular pH and temperature is crucial for understanding physiological activities.
• Existing methods lack the capability for real-time, dual-parameter imaging at the cellular level.
• Investigating cellular metabolism requires tools that can track concurrent changes in pH and temperature.

Purpose of the Study:

• To develop a novel organic small-molecule fluorescent probe, RH-Co, for simultaneous pH and temperature imaging.
• To investigate dynamic changes in cellular pH and temperature during metabolic processes.
• To utilize RH-Co for real-time monitoring of cellular responses to stimuli.

Main Methods:

• Development of RH-Co, an organic small-molecule fluorescent probe.
• Characterization of RH-Co's pH response (pKa ≈ 7.14) and temperature sensitivity (3.12%/°C at 37 °C).
• Simultaneous dynamic imaging of intracellular pH and temperature in living cells using two fluorescent channels.

Main Results:

• RH-Co successfully achieved simultaneous dynamic monitoring of pH and temperature in living cells.
• Observed significant intracellular pH decrease with temperature increase post-37 °C.
• Demonstrated correlation between glucose-induced ATP synthesis, temperature rise, and pH decrease.
• Showcased RH-Co's ability to track pH and temperature changes during FCCP stimulation, copper metabolism, and autophagy.

Conclusions:

• RH-Co is an effective tool for simultaneous dynamic imaging of cellular pH and temperature.
• The probe facilitates the study of metabolic regulation and cellular responses.
• RH-Co provides insights into the interplay between pH, temperature, and cellular activities like energy metabolism and autophagy.