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Intracellularly trapped pH indicators.

J A Thomas

    Society of General Physiologists Series
    |January 1, 1986
    PubMed
    Summary
    This summary is machine-generated.

    The trapped indicator technique offers a non-destructive method for real-time intracellular pH monitoring using standard lab equipment. While leakage can be an issue, strategies like using BCECF or perfusion minimize this, enabling sensitive pH change detection.

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    Area of Science:

    • Cellular Biology
    • Biochemistry
    • Physiology

    Background:

    • Intracellular pH (pH) is a critical regulator of cellular processes.
    • Accurate monitoring of intracellular pH is essential for understanding cell function.
    • Existing methods may have limitations in sensitivity, real-time capability, or cell integrity.

    Purpose of the Study:

    • To evaluate the trapped indicator technique for intracellular pH monitoring.
    • To highlight the advantages and limitations of this method.
    • To provide strategies for optimizing intracellular pH measurements.

    Main Methods:

    • Utilizing fluorescence or absorbance measurements with trapped indicators.
    • Employing fluorescein dyes for pH sensing.
    • Distinguishing between cytoplasmic and mitochondrial pH using specific indicators (CF and F).

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    Main Results:

    • The technique is non-destructive and allows for continuous, real-time monitoring with high sensitivity (0.01 pH change).
    • Leakage of indicators, particularly F, is a significant challenge requiring correction.
    • Strategies such as using BCECF, lowering incubation temperature, or perfusion can minimize leakage.

    Conclusions:

    • The trapped indicator technique is a versatile and sensitive method for intracellular pH monitoring.
    • Careful selection of indicators and optimization of experimental conditions are crucial for accurate results.
    • Further research is needed to understand the generality of cytoplasmic acidification induced by ester hydrolysis.