Synchronization-based image reconstruction for three-dimensional wide-field confocal imaging of periodically moving objects beyond the frame rate

TL;DR

This paper presents an extension of a confocal imaging reconstruction method that enables 3D and 2D imaging of fast periodic motions beyond frame rate limits, using synchronization without extra hardware.

Contribution

The authors extend their previous method to 3D and 2D wide-field imaging, allowing high-speed imaging of periodic objects with standard confocal microscopes through synchronization.

Findings

Successfully visualized 100 Hz oscillating particles in 3D.

Accurately captured periodic particle dynamics.

No additional hardware required for implementation.

Abstract

We extend our previously proposed image reconstruction method, which allows confocal microscopes to capture periodically moving objects at frequencies beyond their frame rates, to three-dimensional and two-dimensional wide-field imaging. This extension is achieved by implementing a synchronization scheme between a confocal laser scanning microscope and a function generator to ensure consistent initial phase alignment across image sequences acquired at different focal depths or fields of view. The method was demonstrated by visualizing the three-dimensional motion of silica particles attached to an aluminum bar oscillating at 100 Hz and the two-dimensional wide-field response of colloidal particles subjected to periodic pulsed excitation. Quantitative single-particle analysis confirmed that the reconstructed images accurately captured the underlying particle dynamics. The extended approach requires no additional specialized hardware and can be readily integrated with conventional confocal microscopes. Thus, it extends the applicability of confocal imaging to the fast dynamics of periodic processes in biological and soft-matter systems.