Neuronal activity in the R2 neurons of the ellipsoid body: CaLexA measurements
This protocol is extracted from research article:
Most sleep does not serve a vital function: Evidence from Drosophila melanogaster
Sci Adv, Feb 20, 2019; DOI: 10.1126/sciadv.aau9253

Animals were grown and treated in the same conditions as in behavioral experiments. After a sleep deprivation of 0.5, 5.5, or 9.5 days, animals were anesthetized on ice, and brains were dissected in phosphate buffer and fixed with 4% paraformaldehyde, as previously described (58). For the quantification of green fluorescent protein (GFP), fly brains were labeled with anti-GFP (1:300; ab290, Abcam). Images were taken under ×400 magnification using a Leica SP8 inverted scanning confocal microscope in the Facility for Imaging by Light Microscopy (FILM), Imperial College London. Data were analyzed using Fiji/ImageJ (59). The measurement of signal intensities was performed as previously described (20).

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