Protocol to record and quantify the intracellular pH in contracting cardiomyocytes

Area of Science:

• Cardiology
• Cell Physiology
• Biophysics

Background:

• Intracellular pH (pH_i) is crucial for regulating cardiac function.
• Cardiac pH_i measurement techniques have advanced significantly since the 1960s.
• Previous methods lacked the resolution and capabilities for dynamic cellular analysis.

Purpose of the Study:

• To present a detailed protocol for measuring intracellular pH (pH_i) in mouse cardiomyocytes.
• To enable simultaneous quantification of pH_i and sarcomere length in contracting cardiomyocytes.
• To highlight the advantages of fluorescent microscopy for cardiac cell research.

Main Methods:

• Isolation of mouse cardiomyocytes.
• Utilization of fluorescent microscopy for pH_i measurement.
• Simultaneous recording of pH_i and sarcomere length in contracting cells.

Main Results:

• Successful isolation of viable mouse cardiomyocytes.
• High-resolution, simultaneous measurement of pH_i and sarcomere dynamics achieved.
• Demonstrated feasibility of the protocol for contracting cardiomyocyte analysis.

Conclusions:

• Fluorescent microscopy provides superior spatial and temporal resolution for cardiac pH_i studies.
• The described method allows for detailed investigation of pH_i regulation in contracting cardiomyocytes.
• This protocol advances the study of cardiac electrophysiology and cell mechanics.