摘要
目的观察体外培养的蓝氏贾第鞭毛虫滋养体超微结构,探究贾第虫滋养体的核仁。方法用改良TYI-S-33培养基培养蓝氏贾第鞭毛虫滋养体,姬姆萨(Giemsa)染色和铁苏木素染色后光镜观察虫体的形态结构;以透射电镜观察虫体的超微结构;以核仁特异性抗体作用虫体产生间接免疫荧光,用激光共聚焦显微镜检测核仁。结果光学显微镜下观察姬姆萨染色后的贾第虫胞质为淡监色,可见1对细胞核,核内有分散的染色体,数目为10条;4对鞭毛、1对半月形的中体、轴柱。铁苏木素染色可见1对椭圆形的泡状细胞核内各有1个细小的核仁。透射电镜观察虫体外观颜面状,呈倒置的梨形:质膜下可见单层排列、大小均匀的小囊泡;可见虫体的细胞器:游离核糖体颗粒、内质网;细胞骨架:腹吸盘、鞭毛、基体、中体的微管结构;核结构:核膜、核孔、核仁、染色体。激光共聚焦显微镜观察发现分裂间期虫体细胞核内有绿色荧光并聚集为核仁,在分裂期虫体内绿色荧光呈弥散分布。结论光镜和电镜观察结果显示体外蓝氏贾第鞭毛虫滋养体的形态结构复杂,有典型的核仁、染色体及复杂的细胞骨架结构。
Objective The ultrastructure characteristics of trophozoites of G.lamblia were observed in vitro with the purpose of investigating the nucleoli's structure of the organism.Methods G.lamblia was cultivated axenically with a modified TYI-S-33 medium.The morphology of the trophozoites were observed under a light microscope after stained with both Giemsa and iron-hematoxylin.The ultrastructures of the trophozoites were observed under a transmission electronic microscope.The organism were incubated at 37℃ with mouse anti-human nucleoli monoclonal antibody and fluorescein-conjugated goat anti-mouse IgG antibody,and then observed under a confocal microscope.Results Under the light microscope,the cytoplasm of trophozoites stained by Giemsa were colored in light-blue.Four pairs of the flagella were clearly seen.A pair of vacuole-like nuclei with small nucleoli was found in the anterior half of the organism stained by Iron-hemotoxylin.Giardia lamblia under the transmission electron microscopy demonstrated in a greater detail the structure of this organism,such as ribosomes,endoplasmic reticulum,nuclear pore complex,nucle-oli, chromosomes,the microtubules of disc and flagella.Under the confocal microscope,green fluorescence material either concentrated in nuclei or existed diffusely in the cytoplasm of the trophozoite during the period of mitotic division.Conclusions There are very complex structures in Giardia lamblia in vitro.
出处
《热带病与寄生虫学》
2010年第2期63-65,2,共4页
Journal of Tropical Diseases and Parasitology
基金
唐山市科技攻关计划2009(09130242a)
关键词
蓝氏贾第鞭毛虫
滋养体
形态学
透射电子显微镜
间接免疫荧光
激光共聚焦显微术
Gi ar di a l ambl i Tr ophozoi t e
Mor phol ogy
Tr ansmi ss i on el ect r oni c mi c r oscope
l nd i r ect immunof l uor es cence
Confoca l mi croscopy
