Modified dual lifetime referencing method for simultaneous optical determination and sensing of two analytes

Area of Science:

• Analytical Chemistry
• Spectroscopy
• Biophysics

Background:

• Simultaneous detection of multiple analytes is crucial in various scientific fields.
• Existing methods for dual-analyte sensing can be complex and require specialized instrumentation.
• Luminescent probes offer sensitive detection but often face challenges in multiplexed analysis.

Purpose of the Study:

• To develop a novel method for simultaneous fluorometric sensing of two analytes.
• To demonstrate the efficacy of the modified dual lifetime referencing (m-DLR) technique.
• To showcase the method's applicability for detecting gases like oxygen and carbon dioxide.

Main Methods:

• Utilized a modified dual lifetime referencing (m-DLR) approach with two luminescent indicators.
• Indicators possessed overlapping spectra but significantly different decay times.
• Excited both indicators with a single light source and measured emissions simultaneously.
• Employed frequency domain analysis, referencing phase shifts of short-lived versus long-lived luminescence at two modulation frequencies.

Main Results:

• Successfully demonstrated simultaneous sensing of oxygen and carbon dioxide.
• The m-DLR method requires simple instrumentation and an uncomplicated optical setup.
• Analytical information was derived from phase shift measurements at two modulation frequencies.
• The technique is adaptable for various dual-analyte sensing scenarios.

Conclusions:

• The modified dual lifetime referencing (m-DLR) method provides a robust platform for simultaneous dual-analyte fluorometric sensing.
• The approach is highly versatile and applicable to detecting different combinations of analytes, including gases and biological molecules.
• Future applications may involve sensing oxygen and chlorophyll or other analyte pairs meeting specific spectral and decay time criteria.