Shadow imaging for panoptical visualization of brain tissue in vivo

Yulia Dembitskaya, Andrew K. J. Boyce, Agata Idziak, Atefeh Pourkhalili Langeroudi, Misa Arizono, Jordan Girard, Guillaume Bourdellès, Mathieu Ducros, Marie Sato-Fitoussi, Amaia Ochoa de Amezaga, Kristell Oizel, Stephane Bancelin, Luc Mercier, Thomas Pfeiffer, Roger J. Thompson, Sun Kwang Kim, Andreas Bikfalvi and U. Valentin Nägerl ()
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Yulia Dembitskaya: CNRS UMR 5297 and University of Bordeaux
Andrew K. J. Boyce: CNRS UMR 5297 and University of Bordeaux
Agata Idziak: CNRS UMR 5297 and University of Bordeaux
Atefeh Pourkhalili Langeroudi: CNRS UMR 5297 and University of Bordeaux
Misa Arizono: CNRS UMR 5297 and University of Bordeaux
Jordan Girard: CNRS UMR 5297 and University of Bordeaux
Guillaume Bourdellès: CNRS UMR 5297 and University of Bordeaux
Mathieu Ducros: Université de Bordeaux, CNRS, INSERM, Bordeaux Imaging Center (BIC), UAR 3420, US 4
Marie Sato-Fitoussi: CNRS UMR 5297 and University of Bordeaux
Amaia Ochoa de Amezaga: CNRS UMR 5297 and University of Bordeaux
Kristell Oizel: Université de Bordeaux, INSERM, Bordeaux Institute of Oncology (BRIC), U1312, Bat B2, Allée Geoffroy St Hilaire
Stephane Bancelin: CNRS UMR 5297 and University of Bordeaux
Luc Mercier: CNRS UMR 5297 and University of Bordeaux
Thomas Pfeiffer: CNRS UMR 5297 and University of Bordeaux
Roger J. Thompson: University of Calgary
Sun Kwang Kim: CNRS UMR 5297 and University of Bordeaux
Andreas Bikfalvi: Université de Bordeaux, INSERM, Bordeaux Institute of Oncology (BRIC), U1312, Bat B2, Allée Geoffroy St Hilaire
U. Valentin Nägerl: CNRS UMR 5297 and University of Bordeaux

Nature Communications, 2023, vol. 14, issue 1, 1-13

Abstract: Abstract Progress in neuroscience research hinges on technical advances in visualizing living brain tissue with high fidelity and facility. Current neuroanatomical imaging approaches either require tissue fixation (electron microscopy), do not have cellular resolution (magnetic resonance imaging) or only give a fragmented view (fluorescence microscopy). Here, we show how regular light microscopy together with fluorescence labeling of the interstitial fluid in the extracellular space provide comprehensive optical access in real-time to the anatomical complexity and dynamics of living brain tissue at submicron scale. Using several common fluorescence microscopy modalities (confocal, light-sheet and 2-photon microscopy) in mouse organotypic and acute brain slices and the intact mouse brain in vivo, we demonstrate the value of this straightforward ‘shadow imaging’ approach by revealing neurons, microglia, tumor cells and blood capillaries together with their complete anatomical tissue contexts. In addition, we provide quantifications of perivascular spaces and the volume fraction of the extracellular space of brain tissue in vivo.

Date: 2023
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DOI: 10.1038/s41467-023-42055-2

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