Luminescent-reaction-enabled super-resolution imaging

Area of Science:

• Biophysics
• Microscopy
• Biochemistry

Background:

• Super-resolution microscopy enhances biological understanding but often relies on light excitation, limiting live-cell applications.
• Reaction-excited luminescence (electrochemiluminescence, chemiluminescence, bioluminescence) offers sensitivity but lacks spatiotemporal resolution due to low photon counts.
• Existing methods struggle to balance high resolution with live-cell compatibility and sensitivity.

Purpose of the Study:

• To develop a chemistry-based super-resolution imaging framework that overcomes the limitations of conventional luminescence-based microscopy.
• To achieve high spatiotemporal resolution using reaction-excited luminescence without external light excitation.
• To enable versatile and sensitive live-cell imaging of biological processes.

Main Methods:

• Developed luminescent-reaction-enabled super-resolution imaging via entropy-weighted correlation combined with deconvolution (RIED).
• Implemented a spatiotemporal recording strategy to capture luminescence information content.
• Utilized a reconstruction strategy adapted to reaction-driven photon statistics for super-resolution reconstruction.

Main Results:

• Achieved super-resolution imaging of electrochemiluminescence (ECL), chemiluminescence (CL), and bioluminescence (BL) with approximately 100 nm resolution.
• Demonstrated sensitive imaging of surface proteins and 41-hour ultralong-term live-cell imaging of mitochondrial transfer dynamics.
• Successfully imaged intracellular organelles using the RIED framework.

Conclusions:

• Established a new class of chemistry-enabled, laser-free super-resolution microscopy.
• RIED expands biological imaging versatility by combining high sensitivity, resolution, and live-cell compatibility.
• This approach offers a powerful tool for studying complex biological dynamics at the nanoscale.