Gorgas K, Böck P
Histochemistry. 1976 Nov 19;50(1):17-31. doi: 10.1007/BF00492782.
Semithin sections (Araldite) of mouse adreno-medullary tissue were examined in the light microscope after perfusion fixation with glutaraldehyde, glutaraldehyde/formaldehyde or after freeze-drying followed by a treatment with hot formaldehyde gas. The following methods were employed: (i) aldehyde-induced fluorescence of catecholamines, (ii) Schmorl's ferric ferricyanide reaction, (iii) argentaffin reaction, and (iiii) staining with alkaline lead citrate followed by Timm's silver sulphide reaction. The correspondence of results obtained by the various methods was proven in consecutive sections or by successively applying different methods to identical sections. Four types of primary catecholamine-storing cells were identified. NA1 cells contain cytoplasmic granules up to 0.3 mum in diameter which stain black with ammoniacal silver and display a bright white to yellow fluorescence. NA2 cells show smaller cytoplasmic granules which stain brown with the argentaffin method and give white catecholamine fluorescence. NA3 cells appear yellow-earth after applying the argentaffin reaction and show greenish fluorescence. NA4 cells are hardly identified in the light microscope. These cells are significantly smaller than the above mentioned cells and characterized by a high nucleo-cytoplasmic ratio. They become straw coloured with ammoniacal silver and show greenish fluorescence. The argentaffin reaction was also used to identify these cells in semithin sections of glutaraldehyde/osmium tetroxide fixed material. The fine structure of the various noradrenalin-storing cells was studied in consecutive thin sections. NA1 cells were found to contain two populations of granules, the larger ones measuring between 300 and 350 nm, the smaller ones about 175 nm. The granules in NA2 cells correspond to this latter population (175 nm). NA3 cells contain an uniform granule population with a main diameter of 120 nm. The smallest granules are seen in NA4 cells being in the dimension of 80 nm. Granules in NA1 and NA2 cells show uniformly high density, whereas those in NA3 and NA4 cells display cores of varying density. Granules with moderately dense cores in NA3 and NA4 cells may represent partially emptied sites of noradrenalin storage or dopamin containing particles.
用戊二醛、戊二醛/甲醛灌注固定或冷冻干燥后用热甲醛气体处理后,在光学显微镜下检查小鼠肾上腺髓质组织的半薄切片(环氧树脂包埋)。采用了以下方法:(i)儿茶酚胺的醛诱导荧光法,(ii)施莫尔氏铁氰化铁反应,(iii)嗜银反应,以及(iiii)用碱性柠檬酸铅染色后进行蒂姆氏硫化银反应。通过连续切片或将不同方法相继应用于相同切片,证明了各种方法所得结果的一致性。鉴定出四种主要的儿茶酚胺储存细胞。NA1细胞含有直径达0.3μm的胞质颗粒,用氨银染色呈黑色,并显示亮白色至黄色荧光。NA2细胞显示较小的胞质颗粒,用嗜银法染色呈棕色,并发出白色儿茶酚胺荧光。应用嗜银反应后,NA3细胞呈黄褐色,并显示绿色荧光。在光学显微镜下很难鉴定出NA4细胞。这些细胞明显小于上述细胞,其特征是核质比高。它们用氨银染色后呈稻草色,并显示绿色荧光。嗜银反应也用于在戊二醛/四氧化锇固定材料的半薄切片中鉴定这些细胞。在连续的薄切片中研究了各种去甲肾上腺素储存细胞的精细结构。发现NA1细胞含有两种颗粒群体,较大的颗粒直径在300至350nm之间,较小的颗粒约为175nm。NA2细胞中的颗粒与后一种群体(175nm)相对应。NA3细胞含有均匀的颗粒群体,主要直径为120nm。在NA4细胞中可见最小的颗粒,尺寸为80nm。NA1和NA2细胞中的颗粒显示出均匀的高密度,而NA3和NA4细胞中的颗粒则显示出密度不同的核心。NA3和NA4细胞中具有中等密度核心的颗粒可能代表去甲肾上腺素储存的部分排空部位或含多巴胺的颗粒。