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Brief freezing steps lead to robust immunofluorescence in the Drosophila nervous system

Drosophila melanogaster possesses a complex nervous system, regulating sophisticated behavioral outputs, that serves as a powerful model for dissecting molecular mechanisms underlying neuronal function and neurodegenerative disease. Immunofluorescence techniques provide a way to visualize the spatio...

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Detalles Bibliográficos
Autores principales: Thapa, Ada, Sullivan, Shea M, Nguyen, Minh Q, Buckley, Dominic, Ngo, Vy T, Dada, Austin O, Blankinship, Essence, Cloud, Veronica, Mohan, Ryan D
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Future Science Ltd 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7031821/
https://www.ncbi.nlm.nih.gov/pubmed/31687836
http://dx.doi.org/10.2144/btn-2018-0067
Descripción
Sumario:Drosophila melanogaster possesses a complex nervous system, regulating sophisticated behavioral outputs, that serves as a powerful model for dissecting molecular mechanisms underlying neuronal function and neurodegenerative disease. Immunofluorescence techniques provide a way to visualize the spatiotemporal organization of these networks, permitting observation of their development, functional location, remodeling and, eventually, degradation. However, basic immunostaining techniques do not always result in efficient antibody penetration through the brain, and supplemental techniques to enhance permeability can compromise structural integrity, altering spatial organization. Here, slow freezing of brains is shown to facilitate antibody permeability without loss of antibody specificity or brain integrity. To demonstrate the advantages of this freezing technique, the results of two commonly used permeation methods – detergent-based and partial proteolytic digestion – are compared.