A DNA-Tile-Based Fluorescence Lighting System for Signal-Amplified Imaging at the Single Live-Cell Level

Area of Science:

• Biotechnology
• Molecular Imaging
• Cell Biology

Background:

• Nucleic acid aptamers offer precise targeting and biosafety for live-cell fluorescence imaging.
• Isothermal amplification enhances signal output but faces challenges with aptamer sequence homology and background noise.
• Existing methods struggle with background signal and cellular internalization of fluorescence.

Purpose of the Study:

• To develop a novel DNA tile-based fluorescent labeling (DTFL) system for enhanced live-cell imaging.
• To overcome limitations of high sequence homology and background noise in aptamer-based imaging.
• To achieve sensitive, accurate, and real-time dynamic monitoring of cellular markers.

Main Methods:

• Developed a DTFL system with separate targeting (aptamer-based) and signal-amplified modules.
• Utilized melamine-responsive array assembly on a membrane surface for nanonet-encapsulated imaging.
• Exploited melamine-mediated T-base mismatches for rapid assembly, distinct from Watson-Crick pairing.

Main Results:

• The DTFL system demonstrated superior rapid cellular imaging capabilities.
• Separation of targeting and imaging elements prevented cellular internalization of fluorescence, enhancing signal intensity.
• Achieved improved imaging sensitivity and accuracy in real-time imaging of various living cells.
• Successfully tracked dynamic changes in specific cellular marker expression levels.

Conclusions:

• The DTFL system offers a promising imaging strategy for live-cell analysis with simple design and operation.
• Melamine-mediated T-base mismatches facilitate efficient signal amplification.
• The system provides a sensitive and accurate tool for dynamic monitoring of cellular markers in laboratory settings.