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Imaging Techniques Combined With Enzyme-Based Procedures That Facilitate the Simultaneous Imaging of Nerves, Neurons, Cartilage And Bones During Development In Gambusia

Published online by Cambridge University Press:  02 July 2020

E. Rosa-Molinar
E. Rosa-Molinar*
Affiliation:
Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, NE68178
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Extract

A persistent problem in elucidating the anatomy of the peripheral nervous system has been an inability to stain both myelinated and unmyelinated nerve fibers. To overcome this problem, our laboratory developed two workmg protocols for reliably and differentially labeling and staining the peripheral nervous system and combined them with an enzyme clearing and staining procedure for the simultaneous visualization of bone and cartilage.

One protocol uses anti-acetylated α-tubulin immunohistochemistry to follow the course, peripheral branching, and origin of the ventral spinal nerve innervating the axial musculature and a second uses anterograde and retrograde transport of selectively applied 3000 molecular weight (MW) biotin dextran amines and/or biocytin to identify specific afferent and efferent projections and their cell bodies. Both procedures can be combined with an enzyme clearing and staining procedure for the simultaneous visualization of bone (alizarin red S) and cartilage (alcian blue) in whole-mount preparations.

Type
Applications of Imaging Techniques to the Study of Embryological Development
Information
Microscopy and Microanalysis , Volume 5 , Issue S2: Proceedings: Microscopy & Microanalysis '99, Microscopy Society of America 57th Annual Meeting, Microbeam Analysis Society 33rd Annual Meeting, Portland, Oregon August 1-5, 1999 , August 1999 , pp. 1074 - 1075
Copyright
Copyright © Microscopy Society of America

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References

1.Rosa-Molinar, , et al, Brain Res., Brain Res. Protoc. in press.Google Scholar
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5. E.R-M is supported by NIH (MH18882).Google Scholar