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Predicting the local pH variations of vesicular exocytotic events at chromaffin cells by amperometry (CAT#: STEM-PPA-0017-LJX)

Introduction

Electrochemical monitoring of the exocytosis process is generally performed through amperometric oxidation of the electroactive messengers released by single living cells. Herein, we consider the vesicular release of catecholamines by chromaffin cells. Each exocytotic event is thus detected as a current spike whose morphology (intensity, duration, area, etc.) features the efficiency of the secretion process. However, the electrochemical oxidation of catechols produces quinone derivatives and protons. As a consequence, unless specific mechanisms may be adopted by a cell to regulate the pH near its membrane, the local pH between the cell membrane and the electrode necessarily drops within the electrode-cell cleft. Though this consequence of amperometric detection is generally ignored, it has been investigated in this work through simulation of the local pH drop created during the amperometric recording of a sequence of exocytotic events. This was performed based on frequencies and magnitudes of release detected at chromaffin cells. T




Principle

Amperometric titration is an electrode titration chemical analysis method that indicates the PH of the solution according to the current change of the solution in the electrolytic cell. The theory is based on the combination of two metals can form an electrolytic cell to provide current. Since hydrogen ions are involved in the reaction, the current of the cell indicates the pH value of the solution.

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:
Chromaffin cells

Notes

When measuring pH value, it is necessary to ensure that the bulb of the electrode is completely immersed in the measured medium, so as to obtain more accurate measurement results.