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Low-frequency modulation sensors using nanosecond fluorophores

J R Lakowicz¹, F N Castellano, J D Dattelbaum

¹Department of Molecular Biology & Biophysics, School of Medicine, University of Maryland, Baltimore 21201, USA.

Analytical Chemistry

|December 30, 1998

Summary

This study introduces a novel fluorescence sensing method using a mix of fluorophores. This technique allows accurate analyte concentration determination by measuring emission modulation, simplifying sensor development.

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Area of Science:

Analytical Chemistry
Biochemistry
Physical Chemistry

Background:

Fluorescence sensing is crucial for detecting analytes.
Existing methods often require specific lifetime changes in sensing fluorophores.
A simplified approach to fluorescence sensing is needed.

Purpose of the Study:

To develop a new fluorescence sensing approach.
To enable analyte concentration determination using intensity modulation.
To overcome limitations of existing fluorescence sensing techniques.

Main Methods:

Utilizing a mixture of long-lifetime analyte-insensitive and short-lifetime analyte-sensitive fluorophores.
Modulating the fluorescence emission at low frequencies.
Measuring the fractional fluorescence intensity of the sensing fluorophore.

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Main Results:

The modulation depth directly correlates with the sensing fluorophore's intensity.
This method is applicable to any fluorophore changing intensity with analyte presence.
Demonstrated feasibility with pH and calcium sensing using specific fluorophore mixtures.

Conclusions:

The developed modulation-based fluorescence sensing is a versatile and effective technique.
It simplifies sensor design by not requiring lifetime changes in the sensing fluorophore.
This approach broadens the applicability of fluorescence-based analyte detection.