SignificanceTwo-photon fluorescence microscopy (TPFM) excited by Gaussian beams requires axial tomographic scanning for three-dimensional (3D) volumetric imaging, which is a time-consuming process, and the slow imaging speed hinders its application for in vivo brain imaging. The Bessel focus, characterized by an extended depth of focus and constant resolution, facilitates the projection of a 3D volume onto a two-dimensional image, which significantly enhances the speed of volumetric imaging.AimWe aimed to demonstrate the ability of a TPFM with a sidelobe-free Bessel beam to provide a promising tool for research in live biological specimens.ApproachComparative in vivo imaging was conducted in live mouse brains and transgenic zebrafish to evaluate the performance of TPFM and Bessel-beam-based TPFM. Additionally, an image-difference method utilizing zeroth-order and third-order Bessel beams was introduced to effectively suppress background interference introduced by sidelobes.ResultsIn comparison with traditional TPFM, the Bessel-beams-based TPFM demonstrated a 30-fold increase in imaging throughput and speed. Furthermore, the effectiveness of the image-difference method was validated in live biological specimens, resulting in a substantial enhancement of image contrast. Importantly, our TPFM with a sidelobe-free Bessel beam exhibited robustness against axial displacements, a feature of considerable value for in vivo experiments.ConclusionsWe achieved rapid, high-contrast, and robust volumetric imaging of the vasculature in live mouse brains and transgenic zebrafish using our TPFM with a sidelobe-free Bessel beam.
Visualization of the spatiotemporal dynamics of propagation is fundamental to understanding dynamic processes ranging from action potentials to electromagnetic pulses, the two ultrafast processes in biology and physics, respectively. Here, we demonstrate differentially enhanced compressed ultrafast photography to directly visualize propagations of passive current flows at approximately 100 m/s along internodes from Xenopus laevis sciatic nerves and of electromagnetic pulses at approximately 5×107 m/s through lithium niobate. The spatiotemporal dynamics of both propagation processes are consistent with the results from computational models, demonstrating that our method can span these two extreme timescales while maintaining high phase sensitivity.
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