Chiba T, Hwang B H, Williams T H
Histochemistry. 1976 Oct 22;49(2):95-106. doi: 10.1007/BF00495673.
Inasmuch as precise correlations of light- and electronmicroscopy are crucial for understanding biostructure, it seemed necessary to bring together the advantages of the glyoxylic acid (GA) method (for inducing monoamine fluorescence) and electron microscopy. A combined fluorescence and electron microscope method using GA is introduced. The brain is perfused by 2% GA in Krebs-Ringer bicarbonate buffer (pH 7.0) and this solution is followed by 4% paraformaldehyde containing 0.5% glutaraldehyde in Sorensen's phosphate buffer (pH 7.4). Sections are cut by cryostat or by vitratome and incubated in 2% GA in phosphate buffer (pH 7.0). Using fluorescence microscopy, features of interest are sketched and/or photographed. Afterwards, the same or subsequent section is processed for electron microscopy. Since axons of catecholamine-containing neurons (as well as their perikarya and terminals) are visualized by GA, the recommended procedure expands the range of studies concerning monoamine neurons that can now be carried out effectively.
鉴于光学显微镜和电子显微镜的精确关联对于理解生物结构至关重要，将乙醛酸（GA）法（用于诱导单胺荧光）和电子显微镜的优势结合起来似乎很有必要。本文介绍了一种使用GA的荧光和电子显微镜联合方法。用含有2% GA的 Krebs - Ringer碳酸氢盐缓冲液（pH 7.0）灌注大脑，然后用含有0.5%戊二醛的4%多聚甲醛的索伦森磷酸盐缓冲液（pH 7.4）灌注。切片用低温恒温器或振动切片机切割，并在含有2% GA的磷酸盐缓冲液（pH 7.0）中孵育。使用荧光显微镜，对感兴趣的特征进行素描和/或拍照。之后对同一切片或后续切片进行电子显微镜处理。由于含儿茶酚胺神经元的轴突（以及它们的胞体和终末）可被GA可视化，推荐的方法扩展了现在可以有效开展的关于单胺能神经元的研究范围。
