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INTRODUCTIONMicroinjection of Drosophila embryos is a common technique used by a wide range of investigators, but some applications require a refined strategy for handling embryos. This article outlines the general procedures for microinjection and quantification of aqueous solutions during high-resolution observation of early development in the fly embryo. It also describes the design of suitable support slides for the manipulation of Drosophila embryos under upright and inverted microscopes.

INTRODUCTIONThe quantitative microinjection of drugs, antibodies, toxins, and manipulated RNAs and proteins into Drosophila embryos--the "pharmacological approach"--provides a unique opportunity to analyze cellular functions in the developing embryo, and provides spatial and temporal resolution that is not readily available through genetic studies. These studies require that the observed effects reflect a dose-response relationship so that the data can be accurately interpreted. Quantitative microinjections can be readily achieved with the addition of a fluorescent tracer to the solution to be injected. Analysis of the resulting integrated fluorescent intensity following injection can then be used to determine the volume and hence the concentration of the solution injected. This protocol outlines the procedures for the microinjection and quantification of aqueous solutions during high-resolution observation of early development in the Drosophila embryo.

INTRODUCTIONThe quantitative microinjection of drugs, antibodies, toxins, and manipulated RNAs and proteins into Drosophila embryos--the "pharmacological approach"--provides a unique opportunity to analyze cellular functions in the developing embryo, and provides spatial and temporal resolution that is not readily available through genetic studies. These studies require that the observed effects reflect a dose-response relationship so that the data can be accurately interpreted. Thus, these microinjection approaches require a more refined strategy for handling embryos, and the use of appropriately designed chambers to mount and observe the embryos greatly facilitates analysis of the biological response to a given injected material. This protocol outlines the procedures for collection and preparation of Drosophila embryos for quantitative microinjection.