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Evaluation of the DMO method for measuring intracellular pH.

J A Hinke, M R Menard

    Respiration Physiology
    |April 1, 1978
    PubMed
    Summary

    This study determined intracellular pH in muscle fibers using two methods: weak acid distribution (5,5-dimethyl-2,4-oxazolidine dione) and a pH microelectrode. Results showed comparable intracellular pH values, even with imperfect uptake data.

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    Area of Science:

    • Physiology
    • Biophysics

    Background:

    • Intracellular pH (pHi) is critical for cellular function.
    • Accurate measurement of pHi in single cells, like muscle fibers, presents challenges.

    Purpose of the Study:

    • To compare intracellular pH measurements in Balanus nubilus muscle fibers using the weak acid distribution method and a glass pH microelectrode.
    • To evaluate the reliability of the weak acid distribution method, specifically 5,5-dimethyl-2,4-oxazolidine dione (DMO), even with suboptimal uptake data.

    Main Methods:

    • Intracellular pH was measured in single muscle fibers of Balanus nubilus.
    • Two techniques were employed: the weak acid distribution method using [14C]DMO and [3H]inulin, and a glass pH microelectrode.
    • Fibers were equilibrated in normal, acidic, or alkaline Ringer solutions.

    Main Results:

    • Calculated intracellular pH values from the DMO method showed favorable agreement with microelectrode measurements.
    • Extrapolating [14C]DMO and [3H]inulin uptake curves to time zero improved the correlation between the two methods.
    • The Waddell and Butler equation was analyzed to understand the accuracy of pHi determination from imperfect uptake data.

    Conclusions:

    • The weak acid distribution method using DMO can provide reliable intracellular pH measurements in muscle fibers, even with non-ideal uptake kinetics.
    • Extrapolation techniques enhance the accuracy of the DMO method for pHi determination.
    • The study validates the utility of the DMO method for physiological research on intracellular pH homeostasis.

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