Vávra J, Dahbiová R, Hollister W S, Canning E U
Department of Parasitology, Charles University, Prague, Czech Republic.
Folia Parasitol (Praha). 1993;40(4):267-72.
Conditions for the effective fluorescence labelling of microsporidian spores by optical brighteners, based on the presence of chitin in the spore wall, are described. Spores of Vairimorpha ephestiae, V. necatrix, V. plodiae, Nosema bombycis, N. apis, N. algerae, Encephalitozoon cuniculi and Enterocytozoon bieneusi were examined. The degree of binding of Calcofluor White M2R (CFW) to untreated spores depends on the conditions and time of storage and the degree of bacterial contamination of the spore sample. Unpurified spores, stored in water are unreliable as control material for the estimation of CFW labelling. However, spores subjected to alkaline treatment by NaOH before CFW application are visualized with ease under all experimental conditions by their bright, not quenching fluorescence in shortwave light (approximately 350 nm) in CFW dilutions of 10(-4) or even lower. Similar improvement in labelling is achieved by exposing spores to CFW dissolved in 0.5-1N NaOH. As well as Calcofluor White M2R other optical brighteners (e.g. Uvitex 2B, Ciba-Geigy or Rylux BA, Ostacolor) can be used for labelling of spores.
基于孢子壁中几丁质的存在，描述了用荧光增白剂对微孢子虫孢子进行有效荧光标记的条件。对埃氏蜜蜂微孢子虫、毁灭蜜蜂微孢子虫、粉斑螟微孢子虫、家蚕微孢子虫、蜜蜂微孢子虫、阿尔及利亚微孢子虫、兔脑炎微孢子虫和比氏肠微孢子虫的孢子进行了检测。荧光增白剂M2R（CFW）与未处理孢子的结合程度取决于储存条件和时间以及孢子样品的细菌污染程度。未纯化且储存在水中的孢子作为CFW标记评估的对照材料不可靠。然而，在CFW应用前用NaOH进行碱性处理的孢子，在所有实验条件下，于10（-4）或更低的CFW稀释液中，在短波光（约350nm）下发出明亮而非淬灭的荧光，从而易于观察。通过将孢子暴露于溶解在0.5 - 1N NaOH中的CFW，也能实现类似的标记改进。除了荧光增白剂M2R外，其他荧光增白剂（如Uvitex 2B、汽巴 - 嘉基公司产品或Rylux BA、Ostacolor）也可用于孢子标记。
