Sarvel A K, Kusel J R, Araújo N, Coelho Pmz, Katz N
Laboratório de Esquistossomose, Centro de Pesquisas René-Rachou-Fiocruz, 31190-002 Belo Horizonte, MG, Brazil.
Mem Inst Oswaldo Cruz. 2006 Sep;101 Suppl 1:289-92. doi: 10.1590/s0074-02762006000900045.
Schistosoma mansoni eggs are classified, according to morphological characteristics, as follows: viable mature and immature eggs; dead mature and immature eggs, shells and granulomas. The scope of this study was to compare the staining characteristics of different morphological types of eggs in the presence of fluorescent labels and vital dyes, aiming at differentiating live and dead eggs. The eggs were obtained from the intestines of infected mice, and put into saline 0.85%. The fluorescent labels were Hoechst 33258 and Acridine Orange + Ethidium Bromide and vital dyes (Trypan Blue 0.4% and Neutral Red 1%). When labelled with the probe Hoechst 33258, some immature eggs, morphologically considered viable, presented fluorescence (a staining characteristic detected only in dead eggs); mature eggs did not present fluorescence, and the other types of dead eggs, morphologically defined, showed fluorescence. As far as Acridine Orange + Ethidium Bromide are concerned, either the eggs considered to be live, or the dead ones, presented staining with green color, and only the hatched and motionless miracidium was stained with an orange color. Trypan Blue was not able to stain the eggs, considered to be dead but only dead miracidia which had emerged out of the shell. Neutral Red stained both live and dead eggs. Only the fluorescent Hoechst 33258 can be considered a useful tool for differentiation between dead and live eggs.
存活的成熟卵和未成熟卵;死亡的成熟卵和未成熟卵、卵壳和肉芽肿。本研究的范围是比较在荧光标记物和活体染料存在的情况下不同形态类型虫卵的染色特征,旨在区分活卵和死卵。虫卵取自感染小鼠的肠道,并置于0.85%的生理盐水中。荧光标记物为Hoechst 33258、吖啶橙+溴化乙锭,活体染料为(0.4%的台盼蓝和1%的中性红)。用探针Hoechst 33258标记时,一些形态学上被认为存活的未成熟卵呈现荧光(一种仅在死卵中检测到的染色特征);成熟卵未呈现荧光,而其他形态学定义的死卵类型则呈现荧光。就吖啶橙+溴化乙锭而言,无论是被认为是活的卵还是死卵,均呈现绿色染色,只有孵化出且不动的毛蚴被染成橙色。台盼蓝无法对被认为是死的卵进行染色,但只能对已从卵壳中孵出的死毛蚴进行染色。中性红对活卵和死卵均有染色。只有荧光Hoechst 33258可被视为区分死卵和活卵的有用工具。