Grant Lester D, Stumpf Walter E
Department of Anatomy and Biological Sciences Research Center, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27514.
J Histochem Cytochem. 1981 Jan;29(1A_suppl):175-180. doi: 10.1177/29.1A_SUPPL.7053241.
A combined technique of formaldehyde-induced fluorescence (FIF) and autoradiography is described for the localization of radioactively labeled substances in relation to monoamine neurons. This method permits the simultaneous visualization of 3H-labeled steroid hormone or drug uptake sites and fluorescing monoamine neural elements (cell bodies, fiber projections, terminals) in the same tissue section. Thin frozen sections cut in a cryostat are freeze-dried, exposed to formaldehyde vapor at 80°C, and carried through dry-mount autoradiography processing steps before fluorescence microscopy screening. Subsequent histological staining of sections and light microscopy are employed for conventional autoradiogram screening. With this procedure, 3H-estradiol and 3H-dihydro-testosterone are localized in various catecholamine (CA) neurons in the diencephalon and lower brain stem of the rat. Also, catecholaminergic as well as noncatechol-aminergic sex steroid target neurons are seen to be innervated by CA terminals in various rat brain regions.
本文描述了一种将甲醛诱导荧光（FIF）与放射自显影相结合的技术，用于定位与单胺能神经元相关的放射性标记物质。该方法能够在同一组织切片中同时显示³H标记的甾体激素或药物摄取位点以及发出荧光的单胺能神经元件（细胞体、纤维投射、终末）。在低温恒温器中切取的薄冰冻切片经冷冻干燥，在80°C下暴露于甲醛蒸汽中，然后在荧光显微镜筛选前进行干式放射自显影处理步骤。随后对切片进行组织学染色和光学显微镜检查，用于常规放射自显影片筛选。通过该程序，³H-雌二醇和³H-二氢睾酮在大鼠间脑和脑干下部的各种儿茶酚胺（CA）神经元中被定位。此外，在大鼠的各个脑区中，可见儿茶酚胺能以及非儿茶酚胺能性类固醇靶神经元接受CA终末的支配。
