The brain is arguably the most complex organ. The branched and extended morphology of nerve cells, their subcellular complexity, the multiplicity of brain cell types as well as their intricate connectivity and the scattering properties of brain tissue present formidable challenges to the understanding of brain function. Neuroscientists have often been at the forefront of technological and methodological developments to overcome these hurdles to visualize, quantify and modify cell and network properties. Over the last few decades, the development of advanced imaging methods has revolutionized our approach to explore the brain. Super-resolution microscopy and tissue imaging approaches have recently exploded. These instrumentation-based innovations have occurred in parallel with the development of new molecular approaches to label protein targets, to evolve new biosensors and to target them to appropriate cell types or subcellular compartments. We review the latest developments for labelling and functionalizing proteins with small localization and functionalized reporters. We present how these molecular tools are combined with the development of a wide variety of imaging methods that break either the diffraction barrier or the tissue penetration depth limits. We put these developments in perspective to emphasize how they will enable step changes in our understanding of the brain.
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The authors acknowledge the critical suggestions of A. Getz, R. Galland and C. Butler with regard to the manuscript. They express their warmest thanks to the many outstanding members of their team and collaborators who participated in elaboration of the concepts in this Review. This work is currently supported by funding from the Ministère de l’Enseignement Supérieur et de la Recherche, Centre National de la Recherche Scientifique, European Research Council grant number 787340 Dyn-Syn-Mem, LabEx BRAIN ANR-10-LABX-43, ANR-10-IDEX-03-02, ANR-16-CE13-0018, ANR-16-CE16-0026-01 and the Conseil Régional de Nouvelle Aquitaine.
The authors declare no competing interests.
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- Quantum dots
Semiconductor particles a few nanometres in size having optical and electronic properties that differ from those of larger particles due to quantum mechanics.
Diamond nanoparticles smaller than 1 µm.
The photochemical alteration of a dye or a fluorophore molecule such that it is permanently unable to fluoresce.
Modification of the structure of a compound by light, especially when accompanied by a change in function.
- Chromophore maturation
The post-translational process through which the chromophore of fluorescent proteins is formed.
A class of photoreceptors in plants, bacteria and fungi used to detect light.
Single-domain antibody fragments consisting of a monomeric variable antibody domain.
Oligonucleotide or peptide molecules that bind to a specific target molecule.
- Galvanometric mirror
An ammeter that indicates it has sensed an electric current by deflecting a light beam with a mirror and is used in laser scanning microscopy.
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Choquet, D., Sainlos, M. & Sibarita, JB. Advanced imaging and labelling methods to decipher brain cell organization and function. Nat Rev Neurosci 22, 237–255 (2021). https://doi.org/10.1038/s41583-021-00441-z
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