Shimosako Nana, Hadjieconomou Dafni, Salecker Iris
Division of Molecular Neurobiology, MRC National Institute for Medical Research, London, UK.
Methods Mol Biol. 2014;1082:57-69. doi: 10.1007/978-1-62703-655-9_4.
Visualization of single neurons within their complex environment is a pivotal step towards uncovering the mechanisms that control neural circuit development and function. This chapter provides detailed technical information on how to use Drosophila variants of the mouse Brainbow-2 system, called Flybow, for stochastic labeling of cells with different fluorescent proteins in one sample. We first describe the genetic strategies and the heat shock regime required for induction of recombination events. This is followed by a detailed protocol as to how to prepare samples for imaging. Finally, we provide specifications to facilitate multichannel image acquisition using confocal microscopy.
在复杂环境中对单个神经元进行可视化，是揭示控制神经回路发育和功能机制的关键一步。本章提供了详细的技术信息，介绍如何使用小鼠Brainbow-2系统的果蝇变体Flybow，在一个样本中用不同荧光蛋白对细胞进行随机标记。我们首先描述诱导重组事件所需的遗传策略和热休克方案。接下来是关于如何制备成像样本的详细方案。最后，我们提供了使用共聚焦显微镜进行多通道图像采集的规格说明。
