Abstract

Laser-scanning confocal fluorescence microscopy is an essential method for biological and medical studies due to its high spatial resolution in three dimensions. Over the last decade, continuous efforts have been made toward the development of techniques for higher-speed confocal fluorescence microscopy in order to observe fast dynamics in biological tissues on the sub-millisecond time scale or to perform high-throughput imaging flow cytometry [1]. Recently, a few techniques based on frequency-division multiplexing (FDM) have been reported to increase the frame rate of confocal fluorescence microscopy [2–4]. The fluorescence imaging using radiofrequency-tagged emission (FIRE) technique is one of the emerging techniques that provides confocal fluorescence image acquisition at an unprecedented frame rate of more than 4,000 fps [4]. However, the frame rates of these techniques are still limited by the scan rate of the mechanical scanning device employed in their systems, such as a resonant scanner or polygon scanner. Here we propose and experimentally demonstrate a method to overcome the limitation in frame rate in FDM-based confocal microscopy.

© 2016 Japan Society of Applied Physics, Optical Society of America