Measurement of Rapid Amiloride-Dependent pH Changes at the Cell Surface by a Proton-Sensitive Field-Effect Transistor (CAT#: STEM-PPA-0003-LJX)

Introduction

The service uses a novel method for the rapid measurement of pH fluxes at close proximity to the surface of the plasma membrane in mammalian cells using an ion-sensitive field-effect transistor (ISFET). In conjuction with an efficient continuous superfusion system, the ISFET sensor was capable of recording rapid changes in pH at the cells' surface induced by intervals of ammonia loading and unloading, even when using highly buffered solutions. Furthermore, the system was able to isolate physiologically relevant signals by not only detecting the transients caused by ammonia loading and unloading, but display steady-state signals as would be expected by a proton transport-mediated influence on the extracellular proton-gradient.

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Principle Applications Procedure Materials Notes Related Products

Principle

The ion-sensitive field-effect transistor method (ISEFT) uses voltage source and drain (source and channel) transistors separated by insulators. Insulators (gates) are made of metal oxides. Hydrogen ions in the medium can accumulate on it, as on a glass electrode, and the positive charge generated by its outer layer will form a "mirror symmetry" in the inner layer of the gate, resulting in a negative charge, so that the gate has electrical conductivity. The lower the pH of the solution, the more hydrogen ions are concentrated on the gate, and the more current flowing between the source and drain can be measured.

Applications

For determining the approximate PH value of solutions in food production, wastewater treatment, pharmaceutical and chemical industries

Procedure

1. Select the "pH" file of the instrument, immerse the cleaned electrode into the standard pH buffer solution to be measured, press the measurement button, turn the positioning adjustment knob, so that the pH value displayed by the instrument is stable at the pH value of the standard buffer solution
2. Release the measuring button, take out the electrode, rinse with distilled water several times, and carefully blot the solution on the electrode with filter paper
3. Place the electrode in the liquid to be tested, press the measurement button, read the stable pH value, and record
4. Release the measurement button, remove the electrode, clean according to step 2, and continue to measure the next sample solution
5. After measuring, clean the electrode and soak the glass electrode in distilled water

Materials

• Sample Type:
Mammalian cells

Notes

1. If you need to measure pH with high accuracy, in order to avoid the effect of CO2 in the air, especially the pH of alkaline solutions, the exposure time to the air should be as short as possible, and the reading should be as fast as possible.
2. In order to ensure the accuracy of the results, the electrodes need to be cleaned in time before and after each measurement.

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