Cornelissen A W, Overdulve J P, Van der Ploeg M
Parasitology. 1984 Feb;88 ( Pt 1):13-25. doi: 10.1017/s0031182000054305.
Feulgen-pararosaniline (SO2) staining was performed on stages in the life-cycle of Isospora (Toxoplasma) gondii, Eimeria tenella, Sarcocystis cruzi and Plasmodium berghei. The fluorescence emission of the stained DNA in nuclei of these stages was examined and compared with absorption microscopy measurements at 560 nm (green light) of the same specimens. Accurate identification of single cells, and especially discrimination between young schizonts and young gamonts was difficult after Feulgen staining. To overcome this problem preparations were first stained with Giemsa and the cells of interest precisely located with the aid of an England finder. The same preparations were then hydrolysed and stained with Feulgen-pararosaniline (SO2), after which the previously identified cells were investigated. The DNA distribution after Feulgen staining corresponded with the shape of nuclei after Giemsa staining. DNA was present in all investigated stages of the four parasites, including macrogamonts of I. (T.) gondii and E. tenella and peripheral blood gamonts of P. berghei. Macrogamonts could be recognized, even at a stage at which they can hardly be differentiated from young schizonts in Giemsa-stained preparations, by their typical distribution pattern of Feulgen fluorescence. Feulgen fluorescence was more granular and confined to the peripheral region of the nucleus, leaving a distinct nucleolus unstained. The horseshoe-shaped nuclei typical of macrogamonts could also be observed in some second generation merozoites of E. tenella, indicating that these merozoites are already sexually differentiated. The relationship between the present cytochemical observations and the ultrastructure of the four investigated parasites is discussed.
对刚地弓形虫(等孢球虫属)、柔嫩艾美耳球虫、克鲁斯肉孢子虫和伯氏疟原虫生命周期各阶段进行了福尔根-副蔷薇苯胺(SO₂)染色。检测了这些阶段细胞核中染色DNA的荧光发射,并与相同标本在560nm(绿光)下的吸收显微镜测量结果进行了比较。福尔根染色后,准确识别单个细胞,尤其是区分年轻裂殖体和年轻配子体很困难。为克服这一问题,先将标本用吉姆萨染色,借助寻像器精确找到感兴趣的细胞。然后将相同的标本进行水解,并用福尔根-副蔷薇苯胺(SO₂)染色,之后对之前识别的细胞进行研究。福尔根染色后的DNA分布与吉姆萨染色后的细胞核形状相对应。在这四种寄生虫的所有研究阶段均存在DNA,包括刚地弓形虫和柔嫩艾美耳球虫的大配子体以及伯氏疟原虫的外周血配子体。即使在吉姆萨染色标本中难以与年轻裂殖体区分的阶段,大配子体也可通过其典型的福尔根荧光分布模式识别出来。福尔根荧光更呈颗粒状,局限于细胞核的周边区域,使明显的核仁未被染色。在柔嫩艾美耳球虫的一些第二代裂殖子中也可观察到大配子体典型的马蹄形细胞核,这表明这些裂殖子已经发生了性分化。讨论了目前的细胞化学观察结果与四种被研究寄生虫超微结构之间的关系。