Label-free image scanning microscopy for kHz super-resolution imaging and single particle tracking

Area of Science:

• Optics and Photonics
• Microscopy
• Biophysics

Background:

• Label-free microscopy offers advantages for biological imaging by avoiding exogenous labels.
• Image scanning microscopy (ISM) provides enhanced resolution and background rejection.
• High-speed imaging is crucial for observing dynamic biological processes and tracking nanoparticles.

Purpose of the Study:

• To modify a label-free image scanning microscope (ISM) for high-speed asynchronous 2D imaging.
• To achieve optical super-resolution imaging with kHz frame rates.
• To demonstrate the system's capability for nanoparticle tracking and live cell imaging.

Main Methods:

• Implemented a resonant mirror for 12kHz one-directional scanning.
• Utilized a chromatic line for instantaneous scanning in the orthogonal direction.
• Adapted optical photon reassignment for super-resolution in the kHz scanning regime.

Main Results:

• Achieved asynchronous 2D imaging at frame rates up to 24kHz.
• Maintained the lateral resolution gain and background rejection of standard ISM.
• Demonstrated single nanoparticle tracking (20nm gold, 50nm silica) and imaging of motile Lactobacillus with improved resolution.

Conclusions:

• The modified ISM system enables high-speed, label-free, super-resolution imaging.
• This advancement facilitates the study of dynamic nanoscale phenomena and biological processes.
• The system offers a powerful tool for advanced microscopy applications in various scientific fields.