Multiplexed structured illumination super-resolution imaging with lifetime-engineered upconversion nanoparticles.

Area of Science:

• Nanotechnology
• Optical Imaging
• Biophysics

Background:

• Optical multiplexing in nanoscale systems offers high information encoding capacity using nanoparticle lifetime fingerprints.
• Conventional widefield imaging faces limitations due to the optical diffraction limit, impacting decoding accuracy and throughput.
• Achieving super-resolution multiplexing requires nanoparticles that maintain modulated excitation and retain optical fingerprints.

Purpose of the Study:

• To develop a tailor-made multiplexed super-resolution imaging method using lifetime-engineered upconversion nanoparticles.
• To overcome the diffraction limit for improved decoding accuracy and throughput in nanoscale optical multiplexing.
• To enhance information handling capacity for applications in disease diagnostics and security.

Main Methods:

• Utilized lifetime-engineered upconversion nanoparticles for super-resolution imaging.
• Employed structured illumination microscopy to achieve high lateral resolution (185 nm).
• Developed a deep learning algorithm for accurate decoding of multiplexed signals, complementing numeric algorithms.

Main Results:

• Demonstrated bright, uniform, and stable nanoparticle performance under structured illumination.
• Achieved a lateral resolution of 185 nm, significantly below the excitation wavelength's diffraction limit.
• Reported over 93% decoding accuracy for three-channel multiplexing and >60% for potential seven-channel multiplexing.

Conclusions:

• The developed lifetime multiplexing super-resolution method significantly enhances spatial multiplexing capacity.
• This approach resolves nanoparticles within diffraction-limited spots, increasing throughput and information density.
• The technique offers a novel solution for managing increasing information content, disease detection, and security challenges.