Correlation of two-photon in vivo imaging and FIB/SEM microscopy

Journal of Microscopy
L Blazquez-LlorcaJochen Herms

Abstract

Advances in the understanding of brain functions are closely linked to the technical developments in microscopy. In this study, we describe a correlative microscopy technique that offers a possibility of combining two-photon in vivo imaging with focus ion beam/scanning electron microscope (FIB/SEM) techniques. Long-term two-photon in vivo imaging allows the visualization of functional interactions within the brain of a living organism over the time, and therefore, is emerging as a new tool for studying the dynamics of neurodegenerative diseases, such as Alzheimer's disease. However, light microscopy has important limitations in revealing alterations occurring at the synaptic level and when this is required, electron microscopy is mandatory. FIB/SEM microscopy is a novel tool for three-dimensional high-resolution reconstructions, since it acquires automated serial images at ultrastructural level. Using FIB/SEM imaging, we observed, at 10 nm isotropic resolution, the same dendrites that were imaged in vivo over 9 days. Thus, we analyzed their ultrastructure and monitored the dynamics of the neuropil around them. We found that stable spines (present during the 9 days of imaging) formed typical asymmetric contacts with axons, where...Continue Reading

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Citations

Feb 26, 2016·Annual Review of Pathology·Jochen Herms, Mario M Dorostkar
Oct 25, 2016·Trends in Cell Biology·Matthia A KarremanJacky G Goetz
Sep 8, 2016·Frontiers in Synaptic Neuroscience·Isabell Begemann, Milos Galic
Oct 28, 2016·Anatomical Science International·Daisuke KogaTsuyoshi Watanabe
Dec 12, 2017·Frontiers in Cellular Neuroscience·Janosch P Heller, Dmitri A Rusakov
May 29, 2018·Development·Agnes BurelIrina Kolotuev
Dec 25, 2019·Andrology·Manuela Simoni, Daniele Santi
Feb 9, 2017·Acta Neuropathologica Communications·Lidia Blazquez-LlorcaJochen Herms
Dec 21, 2016·Brain Plasticity·Jenea M Bin, David A Lyons
Feb 26, 2019·Journal of Physics D: Applied Physics·Toshio AndoGiovanni Zifarelli
Dec 21, 2018·Frontiers in Neuroanatomy·Giulia BolascoCornelius T Gross
Apr 11, 2021·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Sharada IyerMegha Kumar
May 1, 2021·Cancers·Julian Taranda, Sevin Turcan

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Methods Mentioned

BETA
electron microcopy
light microscopy
electron microscopy
light microcopy
transmission electron microscopy
fluorescence microscopy

Software Mentioned

Fiji
Reconstruct
Zen

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