A Rhodamine-Based Ratiometric Fluorescent Sensor for Dual-Channel Visible and Near-Infrared Emission Detection of...

Area of Science:

• Biochemistry
• Cell Biology
• Biomedical Imaging

Background:

• Intracellular NAD(P)H concentrations are vital for understanding cellular metabolism and redox biology.
• Accurate detection and dynamic monitoring of NAD(P)H are essential for studying physiological and pathological processes.

Purpose of the Study:

• To introduce Sensor A, a near-infrared ratiometric fluorescent sensor for real-time, quantitative imaging of NAD(P)H fluctuations in live cells.
• To validate Sensor A's performance in various cellular and in vivo models.

Main Methods:

• Development of a near-infrared ratiometric fluorescent sensor (Sensor A) combining a 3-quinolinium acceptor and a rhodamine dye.
• Utilizing changes in fluorescence emission at 465 nm and 650 nm upon NAD(P)H binding for ratiometric measurements.
• Validation in HeLa and MD-MB-453 cells, including co-localization studies and in vivo imaging in Drosophila larvae.

Main Results:

• Sensor A demonstrated high sensitivity and specificity for NAD(P)H detection with superior photophysical properties.
• Ratiometric measurements were accurate and minimized interference from environmental factors.
• Validated mitochondrial targeting and successful real-time tracking of NAD(P)H fluctuations in cells and Drosophila larvae under various conditions.

Conclusions:

• Sensor A is a powerful tool for real-time, quantitative imaging of intracellular NAD(P)H.
• It facilitates the exploration of cellular metabolism and redox biology in live cells and organisms.
• Represents a significant advancement in NAD(P)H imaging for biomedical research.