Protocol for tissue clearing and 3D analysis of dopamine neurons in the developing mouse midbrain

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Protocol for tissue clearing and 3D analysis of dopamine neurons in the developing mouse midbrain
 
Adolfs, Youri; Raj, Divya D.A.; Brignani, Sara; Pasterkamp, R. Jeroen
(2021) STAR protocols, volume 2, issue 3
(Article)
Abstract
Advances in tissue clearing enable analysis of complex migratory patterns of developing neurons in whole intact tissue. Here, we implemented a modified version of 3DISCO to study migration of midbrain dopamine (DA) neurons. We provide a detailed protocol starting from whole-brain immunostaining, tissue clearing, and ultramicroscopic imaging to post-acquisition quantification ... read more
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Keywords: Antibody, Microscopy, Model Organisms, Molecular Biology, Neuroscience, General Biochemistry,Genetics and Molecular Biology, General Immunology and Microbiology, General Neuroscience, Journal Article
DOI: https://doi.org/10.1016/j.xpro.2021.100669
ISSN: 2666-1667
Publisher: Cell Press
Note: Funding Information: We thank Dr. Alain Chedotal for training provided to Y.A. We thank Dr. Eljo Y van Battum for help in setting up the 3DISCO protocol at the MIND facility (mindresearchfacility.nl). We thank Roger Koot for IT support. This work was supported by mDANeurodev, FP/2007–2011, Grant 222999; Dutch Top Institute Pharma Project T5-207; Universiteit Utrecht (MIND research facility, Dynamics of Youth); the People Program (Marie Curie Actions) of the European Union's Seventh Framework Program (FP7) 2007–2013 under REA grant agreement number 289581 (NPlast); Netherlands Organization for Scientific Research (ALW-NWO VICI); and Stichting Parkinson Fonds to R.J.P. R.J.P. is funded through the Gravitation program of the Dutch Ministry of Education, Culture, and Science and the Netherlands Organization for Scientific Research. Y.A. D.D.A.R. S.B. and R.J.P. conceived the project; Y.A. set up ultramicroscopy imaging in the MIND facility (MIND research facility; www.mindresearchfacility.nl); Y.A. with help of the other authors, optimized and conducted immunostaining and imaging; Y.A. conceived the quantitative method for Imaris™ analysis with help from D.D.A.R. and S.B.; Y.A. and R.J. wrote the manuscript with help from D.D.A.R. and S.B. The authors declare no competing interests. Funding Information: We thank Dr. Alain Chedotal for training provided to Y.A. We thank Dr. Eljo Y van Battum for help in setting up the 3DISCO protocol at the MIND facility (mindresearchfacility.nl). We thank Roger Koot for IT support. This work was supported by mDANeurodev, FP/2007–2011, Grant 222999; Dutch Top Institute Pharma Project T5-207 ; Universiteit Utrecht (MIND research facility, Dynamics of Youth); the People Program (Marie Curie Actions) of the European Union’s Seventh Framework Program (FP7) 2007–2013 under REA grant agreement number 289581 (NPlast); Netherlands Organization for Scientific Research (ALW-NWO VICI); and Stichting Parkinson Fonds to R.J.P. R.J.P. is funded through the Gravitation program of the Dutch Ministry of Education, Culture, and Science and the Netherlands Organization for Scientific Research . Publisher Copyright: © 2021 The Author(s)
(Peer reviewed)