All-synchronized Picosecond Pulses and Time-gated Detection Improve the Spatial Resolution of Two-photon STED Microscopy in Brain Tissue Imaging

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2023 Aug 24

PMID 37616194

Authors

Hirokazu Ishii

Kohei Otomo

Ching-Pu Chang

Miwako Yamasaki

Masahiko Watanabe

Hiroyuki Yokoyama

Tomomi Nemoto

Affiliations

Soon will be listed here.

Abstract

Super-resolution in two-photon excitation (2PE) microscopy offers new approaches for visualizing the deep inside the brain functions at the nanoscale. In this study, we developed a novel 2PE stimulated-emission-depletion (STED) microscope with all-synchronized picosecond pulse light sources and time-gated fluorescence detection, namely, all-pulsed 2PE-gSTED microscopy. The implementation of time-gating is critical to excluding undesirable signals derived from brain tissues. Even in a case using subnanosecond pulses for STED, the impact of time-gating was not negligible; the spatial resolution in the image of the brain tissue was improved by approximately 1.4 times compared with non time-gated image. This finding demonstrates that time-gating is more useful than previously thought for improving spatial resolution in brain tissue imaging. This microscopy will facilitate deeper super-resolution observation of the fine structure of neuronal dendritic spines and the intracellular dynamics in brain tissue.

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