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Imaging dendritic spines in the hippocampus of a living mouse by 3D-STED microscopy

2023 Score: 40 ? 0–100 AI score estimating relevance to the microplastics field. Papers below 30 are filtered from public browse.
Stéphane Bancelin, Luc Mercier, Johannes L. Roos, Mohamed Belkadi, Thomas Pfeiffer, Sun Kwang Kim, U. Valentin Nägerl

Summary

Researchers extended 3D STED super-resolution microscopy to image dendritic spines in the hippocampus of living mice, achieving nanoscale resolution in three dimensions within deep brain tissue and opening new possibilities for studying synaptic structures in vivo.

Body Systems
Models
Study Type In vivo

Abstract STED microscopy has been used to address a wide range of neurobiological questions in optically well-accessible samples like cell culture or brain slices. However, the application of STED to deeply embedded structures in the brain of living animals remains technically challenging. In previous work, we established chronic STED imaging in the hippocampus in vivo but the gain in spatial resolution was restricted to the lateral plane. In this study, we report on extending the gain in STED resolution into the optical axis to visualize dendritic spines in the hippocampus in vivo . The approach is based on a spatial light modulator to shape the focal STED light intensity in all three dimensions and a conically shaped window that is compatible with an objective that has a long working distance and a high numerical aperture. Moreover, we corrected distortions of the laser wavefront to optimize the shape of the bottle beam of the STED laser, which is required for 3D-STED microscopy. In summary, we present a methodology to improve the axial resolution for STED microscopy in the deeply embedded hippocampus in vivo , facilitating longitudinal studies of neuroanatomical plasticity at the nanoscale in a wide range of (patho-)physiological contexts.

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