EdU and Immunofluorescence Staining of Nothobranchius furzeri Organotypic Cultures

• Published in: Cold Spring Harbor protocols Vol. 2023; no. 3; p. 107790
• Main Authors: Bagnoli, Sara, Terzibasi Tozzini, Eva, Cellerino, Alessandro
• Format: Journal Article
• Language: English
• Published: United States 01-03-2023
• Subjects: DNA; Fluorescent Antibody Technique; Microphysiological Systems; Staining and Labeling
• ISSN: 1559-6095
• DOI: 10.1101/pdb.prot107790

Immunofluorescence is a widely used technique to visualize the localization of proteins of interest. Nucleoside analogs, such as 5-ethynyl-2'-deoxyuridine (EdU), are incorporated into newly synthesized DNA and enable permanent labeling of newly divided cells. Both these techniques can be applied to long-term organotypic culture of
in a fashion similar to that already described for tissue sections. We report here our optimized method for immunofluorescence and EdU staining of
organotypic slices.