National Institutes of Health, National Institute of Child Health and Human Development, 35 Lincoln Drive, msc 3715, Bethesda, MD 20892, USA.
J Neurosci Methods. 2011 Jan 15;194(2):312-5. doi: 10.1016/j.jneumeth.2010.10.020. Epub 2010 Nov 11.
Light and electron microscopy (LM and EM) both offer important advantages for characterizing neuronal circuitry in intact brains: LM can reveal the general patterns neurons trace between brain areas, and EM can confirm synaptic connections between identified neurons within a small area. In a few species, genetic labeling with fluorescent proteins has been used with LM to visualize many kinds of neurons and to analyze their morphologies and projection patterns. However, combining these large-scale patterns with the fine detail available in EM analysis has been a technical challenge. To analyze the synaptic connectivity of neurons expressing fluorescent markers with EM, we developed a dual-labeling method for use with pre-embedded brains. In Drosophila expressing genetic labels and also injected with markers we visualized synaptic connections among two populations of neurons in the AL, one of which has been shown to mediate a specific function, odor evoked neural oscillation.
光镜和电镜（LM 和 EM）在描绘完整大脑中的神经元回路方面都具有重要优势：LM 可以揭示神经元在脑区之间的大致模式，而 EM 可以在小区域内确认已识别神经元之间的突触连接。在少数物种中，利用荧光蛋白的遗传标记与 LM 结合使用，可以可视化多种神经元，并分析它们的形态和投射模式。然而，将这些大规模模式与 EM 分析中提供的精细细节相结合一直是一项技术挑战。为了用电镜分析表达荧光标记的神经元的突触连接，我们开发了一种用于预包埋大脑的双标记方法。在表达遗传标记并注射标记物的果蝇中，我们可视化了 AL 中两个神经元群体之间的突触连接，其中一个群体已被证明介导特定功能，即气味诱发的神经振荡。
