Orthogonal DNA Self-Assembly-Based Expansion Microscopy Platform for Amplified, Multiplexed Biomarker Imaging

Area of Science:

• Biotechnology
• Microscopy
• Molecular Biology

Background:

• Expansion microscopy (ExM) enables nanoscale imaging but suffers from signal loss and low signal-to-noise ratio (SNR).
• Existing ExM methods have limitations in signal amplification and multiplexing capabilities.

Purpose of the Study:

• To develop a novel orthogonal DNA self-assembly-based ExM (o-DAExM) platform.
• To overcome the limitations of conventional ExM, including fluorescence loss and low SNR.
• To enhance signal amplification and multiplexing for super-resolution imaging.

Main Methods:

• Developed an o-DAExM platform utilizing hybridization chain reaction for signal amplification.
• Compared o-DAExM with immunofluorescence-based ExM for cellular cytoskeleton imaging.
• Applied o-DAExM for exosome analysis and multiplexed protein target detection in single cells.

Main Results:

• o-DAExM demonstrated significant signal amplification and enhanced SNR compared to conventional methods.
• Achieved high-resolution imaging of cellular cytoskeleton with outstanding performance and reliability.
• Successfully revealed heterogeneous exosome information and enabled multiplexed protein analysis.

Conclusions:

• o-DAExM provides a straightforward and effective method for nanoscale imaging with improved signal and resolution.
• The platform offers expandable multiplexing capabilities for diverse biological samples.
• o-DAExM presents significant potential for studying nanoscale structures and biological networks.