Here, we present an optimized iDISCO+ protocol combining tissue clearing and light sheet microscopy to map the postnatal development of oxytocin and vasopressin neurons in mouse hypothalamus. We describe tissue preparation, immunostaining, clearing, and imaging. We then detail how to process the 3D cell dataset to analyze cell network using a point-based recording procedure that accurately maps neurons in the Allen brain atlas. This protocol can be applied to any neuronal population, in different brain regions and at different developmental stages. For complete details on the use and execution of this protocol, please refer to Soumier et al. (2021)..
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http://dx.doi.org/10.1016/j.xpro.2022.101968 | DOI Listing |
STAR Protoc
December 2024
University Bordeaux, Inserm, Bordeaux Institute of Oncology (BRIC), UMR 1312, F-33076 Bordeaux, France. Electronic address:
Tissue clearing enables deep imaging of long biological structures using light microscopy approaches. Protocols such as iDISCO are not currently available for human skin. Here, we present Skin-iDISCO, a tissue-clearing and labeling protocol for morphometric analysis of human cutaneous vasculature.
View Article and Find Full Text PDFSci Transl Med
July 2024
Department of Molecular and Cellular Endocrinology, Arthur Riggs Diabetes and Metabolism Research Institute, City of Hope Beckman Research Institute, Duarte, CA 91010, USA.
Five hundred thirty-seven million people globally suffer from diabetes. Insulin-producing β cells are reduced in number in most people with diabetes, but most individuals still have some residual β cells. However, none of the many diabetes drugs in common use increases human β cell numbers.
View Article and Find Full Text PDFSTAR Protoc
March 2023
Institute of Cognitive Science Marc Jeannerod, CNRS, Bron, France; iMIND, Center of Excellence for Autism, le Vinatier Hospital, Bron, France. Electronic address:
Here, we present an optimized iDISCO+ protocol combining tissue clearing and light sheet microscopy to map the postnatal development of oxytocin and vasopressin neurons in mouse hypothalamus. We describe tissue preparation, immunostaining, clearing, and imaging. We then detail how to process the 3D cell dataset to analyze cell network using a point-based recording procedure that accurately maps neurons in the Allen brain atlas.
View Article and Find Full Text PDFSci Rep
April 2022
Max Planck Institute for Molecular Biomedicine, Röntgenstr. 20, 48149, Münster, Germany.
In recent years, 3D cell culture has been gaining a more widespread following across many fields of biology. Tissue clearing enables optical analysis of intact 3D samples and investigation of molecular and structural mechanisms by homogenizing the refractive indices of tissues to make them nearly transparent. Here, we describe and quantify that common clearing solutions including benzyl alcohol/benzyl benzoate (BABB), PEG-associated solvent system (PEGASOS), immunolabeling-enabled imaging of solvent-cleared organs (iDISCO), clear, unobstructed brain/body imaging cocktails and computational analysis (CUBIC), and ScaleS4 alter the emission spectra of Alexa Fluor fluorophores and fluorescent dyes.
View Article and Find Full Text PDFNeuroimage
February 2022
Paul Flechsig Institute of Brain Research, Medical Faculty, University of Leipzig, Liebigstraße 19, Leipzig 04103, Germany; Department of Neurophysics, Max Planck Institute for Human Cognitive and Brain Science, Stephanstraße 1a, Leipzig 04103, Germany. Electronic address:
The accessibility of new wide-scale multimodal imaging techniques led to numerous clearing techniques emerging over the last decade. However, clearing mesoscopic-sized blocks of aged human brain tissue remains an extremely challenging task. Homogenizing refractive indices and reducing light absorption and scattering are the foundation of tissue clearing.
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