Immunofluorescence Staining of <em>Drosophila</em> Brains for the Single-Cell Imaging of Glial Cells

Source: Batelli, S., et al. Application of MultiColor FlpOut Technique to Study High Resolution Single Cell Morphologies and Cell Interactions of Glia in Drosophila. J. Vis. Exp. (2017). This video demonstrates the immunofluorescence staining of a transgenic Drosophila brain for single-cell imaging of glial cells. The brain expresses membrane-targeted proteins fused with different combinations of antigenic epitopes under the control of a glial-specific recombinase system. The isolated and fixed brains are stained with primary antibodies specific to the antigenic epitopes, followed by fluorophore-conjugated secondary antibodies. This labeling enables the simultaneous imaging of different glial cell subpopulations across various brain regions.