Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Search research articles

Related Experiment Videos

Background suppression in frequency-domain fluorometry.

J R Lakowicz¹, I Gryczynski, Z Gryczynski

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

Analytical Biochemistry

|December 28, 1999

Summary

This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Photoselection of Luminescent Molecules in Anisotropic Media in the Case of Two-Photon Excitation. Part II. Experimental Studies of Hoechst 33342 in Stretched Poly(vinyl alcohol) Films.

Zeitschrift fur Naturforschung. A, Journal of physical sciences·2021

Same author

Site-to-site distance distribution in flexible molecules: theoretical evaluation of the donor and/or acceptor fluorescence decay function.

Journal of luminescence·2021

Same author

Correction to: Transamidase site-targeted agents alter the conformation of the transglutaminase cancer stem cell survival protein to reduce GTP binding activity and cancer stem cell survival.

Oncogene·2021

Same author

Time-Resolved Fluorescence Intensity and Anisotropy Decays of 2,5-Diphenyloxazole by Two-Photon Excitation and Frequency-Domain Fluorometry.

The Journal of physical chemistry·2019

Same author

Photophysical properties of 2-Phenylindole in poly (vinyl alcohol) film at room temperature. Enhanced phosphorescence anisotropy with direct triplet state excitation.

Methods and applications in fluorescence·2019

Same author

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

Acta physica Polonica: A·2019

Researchers developed a new method for frequency-domain fluorometry to suppress autofluorescence. This technique uses gated detection to improve measurements of long-lived fluorophores, enabling clearer data analysis.

Area of Science:

Analytical Chemistry
Spectroscopy
Biophysics

Background:

Gated detection is crucial for time-domain fluorometry to eliminate short-lived autofluorescence.
Frequency-domain fluorometry traditionally lacks direct methods for autofluorescence suppression.
Interfering autofluorescence complicates the analysis of long-lived fluorophores in frequency-domain measurements.

Purpose of the Study:

To introduce a direct method for real-time autofluorescence suppression in frequency-domain fluorometry.
To enable accurate measurements of long-lived fluorophores by removing background noise.
To provide a universally applicable technique for frequency-domain fluorescence spectroscopy.

Main Methods:

Utilized a gated detector synchronized with a pulsed excitation light source.

Related Experiment Videos

Introduced a time delay after each excitation pulse for prompt autofluorescence decay.

Employed a long-lifetime standard for reference signal generation under identical conditions.

Main Results:

Demonstrated real-time suppression of autofluorescence in frequency-domain fluorometry.
Showcased that sample and reference signals can be analyzed without additional processing.
Simulations confirmed the method's ability to resolve single and multiexponential decays amidst autofluorescence.

Conclusions:

The developed gated detection method effectively suppresses autofluorescence in frequency-domain fluorometry.
This technique simplifies data analysis and enhances the accuracy of measurements for long-lived fluorophores.
The method offers a significant advancement for fluorescence spectroscopy applications requiring background noise reduction.