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Related Experiment Video

Updated: Jul 10, 2026

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

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

Published on: April 28, 2014

An efficient ratiometric fluorescent probe for tracking dynamic changes in lysosomal pH.

Qianqian Wang¹, Liyi Zhou, Liping Qiu

¹Molecular Sciences and Biomedicine Laboratory, State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University, Changsha 410082, PR China. xbzhang@hnu.edu.cn.

|June 25, 2015

Summary

A new fluorescent probe, FR-Lys, enables tracking of dynamic lysosomal pH changes. This ratiometric probe offers fast, reversible, and selective pH sensing for biological applications.

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Area of Science:

Biochemistry
Cell Biology
Chemical Biology

Background:

Lysosomes are acidic organelles crucial for cellular processes.
Monitoring lysosomal pH dynamics is vital for understanding cell function.
Existing fluorescent probes often lack the ability to track dynamic pH changes.

Purpose of the Study:

To develop a novel lysosome-targeted ratiometric fluorescent probe for dynamic pH monitoring.
To characterize the probe's response to pH variations within the lysosomal range.
To demonstrate the probe's utility in live cell imaging of lysosomal pH.

Main Methods:

Synthesis of a new ratiometric fluorescent probe (FR-Lys) by combining morpholine, a xanthane derivative, and an o-hydroxy benzoxazole group.
Investigating the probe's pH-dependent fluorescence response (4.0-6.3) using spectrophotometry.
Evaluating probe characteristics including response time, reversibility, selectivity, photostability, and cytotoxicity.
Applying the probe for dynamic imaging of lysosomal pH in biological systems.

Main Results:

The FR-Lys probe exhibits a ratiometric fluorescence response to pH changes between 4.0 and 6.3.
The probe demonstrates fast, reversible, and highly selective pH sensing.
FR-Lys shows good photostability and low cytotoxicity.
Successful application in dynamic imaging of lysosomal pH changes in cells.

Conclusions:

The developed FR-Lys probe is effective for real-time monitoring of lysosomal pH.
Its design, incorporating targeting, spirocycle modulation, and ESIPT, enables reliable ratiometric sensing.
FR-Lys offers significant advantages for studying dynamic pH fluctuations in biological environments.