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Anisotropy-based sensing with reference fluorophores.

J R Lakowicz¹, I Gryczynski, Z Gryczynski

¹University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, Maryland, 21201, USA.

Analytical Biochemistry

|February 26, 1999

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ADVANCES IN FLUORESCENCE SPECTROSCOPY: MULTI-PHOTON EXCITATION, ENGINEERED PROTEINS, MODULATION SENSING AND MICROSECOND RHENIUM METAL-LIGAND COMPLEXES.

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This study introduces a novel fluorescence sensing method using steady-state anisotropy measurements with reference fluorophores. This technique allows for adjustable sensitivity and wide dynamic range, applicable in clinical and analytical chemistry.

Area of Science:

Analytical Chemistry
Biophysical Chemistry

Background:

Fluorescence sensing is crucial for various analytical applications.
Traditional methods may have limitations in dynamic range and sensitivity.

Purpose of the Study:

To develop a new fluorescence sensing approach utilizing steady-state anisotropy.
To demonstrate the versatility and adaptability of this anisotropy-based sensing method.

Main Methods:

Measurements of steady-state anisotropies in the presence of reference fluorophores with known anisotropies.
Utilizing the weighted average principle of anisotropies in mixtures.
Employing reference fluorophores to establish adjustable starting anisotropy values (near zero or 0.9).

Main Results:

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Achieved sensitive measurements with a simple light-emitting diode source without an excitation polarizer.

Conclusions:

Anisotropy-based sensing offers a flexible and wide dynamic range approach.
The sensitive range can be tuned by adjusting reference fluorophore intensity.
This method holds significant potential for diverse applications in clinical and analytical chemistry.