Live STED imaging of functional neuroanatomy

Arizono M(1)(2)(3), Idziak A(4)(5), Nägerl UV(6)(7).

Author information:
(1)Interdisciplinary Institute for Neuroscience, University of Bordeaux, CNRS
UMR 5297, Bordeaux, France. .
(2)The Hakubi Center for Advanced Research, Kyoto University, Kyoto, Japan.
.
(3)Department of Pharmacology, Kyoto University Graduate School of Medicine,
Kyoto University, Kyoto, Japan. .
(4)Interdisciplinary Institute for Neuroscience, University of Bordeaux, CNRS
UMR 5297, Bordeaux, France.
(5)Department of Pharmacology, Kyoto University Graduate School of Medicine,
Kyoto University, Kyoto, Japan.
(6)Interdisciplinary Institute for Neuroscience, University of Bordeaux, CNRS
UMR 5297, Bordeaux, France. .
(7)Department of Anatomy and Cell Biology, University Medical Center,
Georg-August-University of Göttingen, Göttingen, Germany.
.

In the mammalian brain, a large network of excitable and modulatory cells
efficiently processes, analyzes and stores vast amounts of information. The
brain’s anatomy influences the flow of neural information between neurons and
glia, from which all thought, emotion and action arises. Consequently, one of
the grand challenges in neuroscience is to uncover the finest structural details
of the brain in the context of its overall architecture. Recent developments in
microscopy and biosensors have enabled the investigation of brain microstructure
and function with unprecedented specificity and resolution, dendritic spines
being an exemplary case, which has provided deep insights into neuronal
mechanisms of higher brain function, such as learning and memory. As
diffraction-limited light microscopy methods cannot resolve the fine details of
brain cells (the ‘anatomical ground truth’), electron microscopy is used instead
to contextualize functional signals. This approach can be quite unsatisfying
given the fragility and dynamic nature of the structures under investigation. We
have recently developed a method for combining super-resolution stimulated
emission depletion microscopy with functional measurements in brain slices,
offering nanoscale resolution in functioning brain structures. We describe how
to concurrently perform morphological and functional imaging with a confocal
STED microscope. Specifically, the procedure guides the user on how to record
astrocytic Ca2+ signals at tripartite synapses, outlining a framework for
analyzing structure-function relationships of brain cells at nanoscale
resolution. The imaging requires 2-3 h and the image analysis between 2 h and 2
d.

© 2025. Springer Nature Limited.

DOI: 10.1038/s41596-024-01132-6
PMID: 40087378

Conflict of interest statement: Competing interests: The authors have no
competing interests as defined by Nature Research, or other interests that might
be perceived to influence the interpretation of the article.

Auteurs Bordeaux Neurocampus