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Brief Freezing Steps Lead to Robust Immunofluorescence in Drosophila Larval Brains

Dominic Buckley, Ada Thapa, Minh Nguyen, Essence Blankinship, Diana Williamson, Veronica Cloud, Ryan D Mohan
doi: https://doi.org/10.1101/319913
Dominic Buckley
University of Missouri - Kansas City
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Ada Thapa
University of Missouri - Kansas City
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Minh Nguyen
University of Missouri - Kansas City
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Essence Blankinship
University of Missouri - Kansas City
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Diana Williamson
University of Missouri - Kansas City
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Veronica Cloud
University of Missouri - Kansas City
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Ryan D Mohan
University of Missouri - Kansas City
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  • For correspondence: mohanrd@umkc.edu
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Abstract

Drosophila melanogaster possess complex neuronal networks regulating sophisticated behavioral outputs that aid in studying the molecular mechanisms of neuronal function and neurodegenerative disease. Immunofluorescence (IF) techniques provide a way to visualize the spatiotemporal organization of these networks, permitting observation of their development, functional location, remodeling, and eventually degradation. However, general immunostaining techniques do not always result in sufficient antibody penetration through the brain, and techniques used to enhance permeability can compromise structural integrity. We have found that freezing larval brains facilitates permeability with no apparent loss of antibody specificity or structural integrity. To demonstrate the advantage of this freezing technique, we compared results to two commonly used permeation methods: Detergent alone (Basic) and proteolytic degradation (Collagenase) techniques.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted May 13, 2018.
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Brief Freezing Steps Lead to Robust Immunofluorescence in Drosophila Larval Brains
Dominic Buckley, Ada Thapa, Minh Nguyen, Essence Blankinship, Diana Williamson, Veronica Cloud, Ryan D Mohan
bioRxiv 319913; doi: https://doi.org/10.1101/319913
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Brief Freezing Steps Lead to Robust Immunofluorescence in Drosophila Larval Brains
Dominic Buckley, Ada Thapa, Minh Nguyen, Essence Blankinship, Diana Williamson, Veronica Cloud, Ryan D Mohan
bioRxiv 319913; doi: https://doi.org/10.1101/319913

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