Williamson W Ryan, Hiesinger P Robin
Department of Physiology and Green Center for Systems Biology, University of Texas Southwestern Medical Center, USA.
J Vis Exp. 2010 Mar 15(37):1936. doi: 10.3791/1936.
The Drosophila brain and visual system are widely utilized model systems to study neuronal development, function and degeneration. Here we show three preparations of the brain and visual system that cover the range from the developing eye disc-brain complex in the developing pupae to individual eye and brain dissection from adult flies. All protocols are optimized for the live culture of the preparations. However, we also present the conditions for fixed tissue immunohistochemistry where applicable. Finally, we show live imaging conditions for these preparations using conventional and resonant 4D confocal live imaging in a perfusion chamber. Together, these protocols provide a basis for live imaging on different time scales ranging from functional intracellular assays on the scale of minutes to developmental or degenerative processes on the scale of many hours.
果蝇的大脑和视觉系统是广泛用于研究神经元发育、功能和退化的模型系统。在这里，我们展示了三种大脑和视觉系统的制备方法，其涵盖了从发育中的蛹期的眼盘-脑复合体到成年果蝇的单个眼睛和大脑解剖的范围。所有方案均针对这些制备物的活细胞培养进行了优化。然而，我们也给出了适用的固定组织免疫组织化学条件。最后，我们展示了使用灌注室中的传统和共振4D共聚焦实时成像对这些制备物进行实时成像的条件。总之，这些方案为在从几分钟尺度的功能性细胞内分析到数小时尺度的发育或退化过程等不同时间尺度上进行实时成像提供了基础。
