A Double Staining Technique using 5-Bromo,4-chloro-3-indolyl-β-D- galactopyranoside (X-gal) and Immunoperoxidase in WholeDrosophilaEmbryos

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Abstract
We have developed a double staining method using 5-bromo,4-chloro-3-indolyl-β-D-galactopyranoside X-gal and immunoperoxidase for whole Drosophila embryos. The dechorionated embryos are fixed in heptane saturated with 4% formaldehyde, then in heptane and 50% methanol. Fixed embryos are devitellinized with a tungsten needle and processed for immunoperoxidase staining immediately prior to peroxidase color development. The embryos are stained with X-gal, then peroxidase staining is resumed. This procedure enables us to observe cells stained with both X-gal and a specific antibody in whole embryos.