Jackson P R, Winkler D G, Kimzey S L, Fisher F M
J Parasitol. 1977 Aug;63(4):593-8.
Samples of rat blood infected with Plasmodium yoelii (3% parasitized erythrocytes), Trypanosoma gambiense, or Trypanosoma equiperdum (greater than 50 trypanosomes per microscope field at 400 X) were fixed with 0.5% glutaraldehyde in phosphate buffered saline, then stained with acridine orange (AO) at 10(-4), 10(-5), or 10(-6) M for 0 to 15 min at 5 C or 25 C and/or ethidium bromide (EB) at 0.05 mg/ml for 20 min at 25 C. Stained cells were analyzed with a laser Cytofluorograf (Bio/Physics Systems, Inc.) to determine if parasites could be detected and differentiated from blood cells by their fluorescent characteristics. Samples of uninfected rat blood with and without leukocytes and P. yoelii-infected blood without leukocytes were treated similarly. In addition, suspensions of T. gambiense and T. equiperdum without all blood cells were stained with AO or EB and analyzed with the Cytofluorograf, as were mixed suspensions of both trypanosome species. EB- but not AO-stained P. yoelii-infected erythrocytes had fluorescent characteristics different from most blood cells. Neither AO- nor EB-stained T. gambiense or T. equiperdum could be differentiated from host blood cells or from each other. The results are discussed with respect to the use of laser flow systems in the detection and analysis of bloodstream dwelling protozoan parasites.
用约氏疟原虫（3%被寄生红细胞）、冈比亚锥虫或马媾疫锥虫（在400倍显微镜视野下每视野大于50个锥虫）感染的大鼠血液样本，用0.5%戊二醛在磷酸盐缓冲盐水中固定，然后分别用10⁻⁴、10⁻⁵或10⁻⁶M的吖啶橙（AO）在5℃或25℃下染色0至15分钟和/或用0.05mg/ml的溴化乙锭（EB）在25℃下染色20分钟。用激光细胞荧光分析仪（生物/物理系统公司）分析染色后的细胞，以确定是否能通过其荧光特性检测到寄生虫并将其与血细胞区分开来。对有无白细胞的未感染大鼠血液样本以及无白细胞的约氏疟原虫感染血液样本进行类似处理。此外，对不含所有血细胞的冈比亚锥虫和马媾疫锥虫悬液用AO或EB染色并用细胞荧光分析仪分析，两种锥虫物种的混合悬液也如此处理。用EB而非AO染色的约氏疟原虫感染红细胞具有与大多数血细胞不同的荧光特性。用AO或EB染色的冈比亚锥虫或马媾疫锥虫均无法与宿主血细胞区分开来，也无法相互区分。讨论了激光流动系统在检测和分析血液中寄生的原生动物寄生虫方面的应用结果。
