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Simultaneous pH Measurement in Endocytic and Cytosolic Compartments in Living Cells using Confocal Microscopy

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Wide range pH measurements using a single H(+)-selective chromoionophore and a time-based flow method.

Afsaneh Safavi¹, Abolfazl Rostamzadeh, Saeed Maesum

¹Department of Chemistry, Faculty of Sciences, Shiraz University, Shiraz 71454, Iran.

|October 31, 2008

Summary

This study introduces a novel pH optode using Nile blue indicator on a triacetyl cellulose membrane. The new time-based flow method allows for pH measurements across a broad 0-10 range, overcoming limitations of traditional steady-state approaches.

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Area of Science:

Analytical Chemistry
Chemical Sensing
Materials Science

Background:

Traditional pH optodes often have a limited linear dynamic range, typically 2-4 pH units.
Current methods for optode use commonly rely on measuring a steady-state response, which can be time-consuming.

Purpose of the Study:

To develop a novel pH optode utilizing a dynamic response approach.
To expand the measurable pH range of optodes beyond conventional limitations.
To introduce a simple, rapid, and cost-effective method for pH determination.

Main Methods:

Immobilization of Nile blue indicator onto a hydrophilic transparent triacetyl cellulose membrane to create a pH optode.
Utilizing the dynamic response of the optode as the analytical signal, deviating from steady-state measurements.
Implementing a time-based flow method for pH measurements.

Main Results:

The developed pH optode demonstrated a significantly expanded linear dynamic range, covering the pH range of 0-10.
The dynamic response method enabled measurements across a much wider pH range compared to traditional steady-state methods.
The procedure proved to be simple, inexpensive, and rapid for pH determination.

Conclusions:

A novel pH optode based on Nile blue and triacetyl cellulose membrane was successfully developed.
The dynamic response in a time-based flow method offers a superior approach for pH sensing with an extended linear range.
This method presents a practical and efficient alternative for broad-range pH measurements.