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Related Experiment Videos

Intracellular pH in single motile cells

J M Heiple, D L Taylor

    The Journal of Cell Biology
    |September 1, 1980
    PubMed
    Summary

    This study measured the cytoplasmic pH of Chaos carolinensis using microfluorometry. The average pH was found to be 6.75, with localized variations and transient drops during wound healing, suggesting pH

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    Area of Science:

    • Cell Biology
    • Biophysics
    • Microscopy

    Background:

    • Understanding cytoplasmic pH is crucial for cellular function.
    • Previous methods for measuring intracellular pH in single-celled organisms were limited.

    Purpose of the Study:

    • To develop and apply a microfluorometric technique for determining cytoplasmic pH in living Chaos carolinensis.
    • To investigate spatial and temporal variations in cytoplasmic pH within individual amebae and during cellular processes like wound healing.

    Main Methods:

    • Microfluorometry using fluorescein-thiocarbamyl (FTC)-ovalbumin as a pH-sensitive probe.
    • In situ calibration of fluorescence ratios against known pH values.
    • Measurement of pH in freely moving amebae, during wound healing, and in different cellular regions.

    Main Results:

    • The average cytoplasmic pH of Chaos carolinensis was determined to be 6.75.
    • Significant spatial variations in pH were observed within single amebae (up to 0.4 U difference).
    • A transient pH drop (up to 0.35 U) occurred at wound sites during healing.

    Conclusions:

    • Cytoplasmic pH in Chaos carolinensis is tightly regulated but exhibits localized heterogeneity.
    • Intracellular pH fluctuations may play a role in regulating cytoplasmic structure and contractility.
    • The developed microfluorometric method provides a reliable tool for studying intracellular pH dynamics.

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