Rickert Christof, Kunz Thomas, Harris Kerri-Lee, Whitington Paul, Technau Gerhard
Institute of Genetics, University of Mainz, Germany.
J Vis Exp. 2013 Mar 4(73):e50150. doi: 10.3791/50150.
In this article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the lipophilic fluorescent membrane marker DiI. This method allows the visualization of neuronal cell morphology in great detail. It is possible to label any cell in the CNS: cell bodies of target neurons are visualized under DIC optics or by expression of a fluorescent genetic marker such as GFP. After labeling, the DiI can be transformed into a permanent brown stain by photoconversion to allow visualization of cell morphology with transmitted light and DIC optics. Alternatively, the DiI-labeled cells can be observed directly with confocal microscopy, enabling genetically introduced fluorescent reporter proteins to be colocalised. The technique can be used in any animal, irrespective of genotype, making it possible to analyze mutant phenotypes at single cell resolution.
在本文中，我们描述了如何通过亲脂性荧光膜标记物DiI的细胞旁注射，对黑腹果蝇胚胎中枢神经系统（CNS）中的神经元进行单独标记。该方法能够极为详细地观察神经元细胞形态。可以标记中枢神经系统中的任何细胞：在微分干涉差（DIC）光学系统下或通过荧光遗传标记物（如绿色荧光蛋白，GFP）的表达来观察目标神经元的细胞体。标记后，通过光转换可将DiI转化为永久性的棕色染色，以便在透射光和DIC光学系统下观察细胞形态。或者，可直接用共聚焦显微镜观察DiI标记的细胞，从而使基因导入的荧光报告蛋白能够共定位。该技术可用于任何动物，无论其基因型如何，从而能够在单细胞分辨率下分析突变体表型。
