Laube U, Kiderlen A F
Robert Koch-Institut, Abteilung Immunologie, Berlin, Germany.
Parasitol Res. 1998 Jul;84(7):559-64. doi: 10.1007/s004360050448.
Even for routine purposes, standard staining of Pneumocystis- or Leishmania-containing materials, e.g., with Giemsa or Diff-Quik, is often unsatisfactory due to poor contrast and to staining of irrelevant structures. In comparison, the bisbenzimide dye Hoechst 33258, a DNA-binding fluorochrome, allows a more precise analysis of such materials. Bisbenzimide stained all stages of these fungal or protozoal organisms with brilliant contrast against a uniformly dark background. The level of background luminescence and staining of detritus or non-DNA structures was very low. Organisms were stained both outside of and within phagocytic cells with equal intensity. Counting of individual microorganisms, e.g., in macrophages heavily parasitized with Leishmania or in Pneumocystis-infected bronchoalveolar lavage, was simplified and more precise. Air-dried cell suspensions, cytocentrifuge preparations, impression smears, or cryocut micrographs showed the advantages of bisbenzimide staining over Diff-Quik. Staining with bisbenzimide could be a valuable auxiliary technique for the analysis of material infected with a variety of microorganisms.
即使是出于常规目的，对含有肺孢子菌或利什曼原虫的材料进行标准染色，例如用吉姆萨染色或Diff - Quik染色，由于对比度差以及无关结构的染色，结果往往不尽人意。相比之下，双苯甲酰亚胺染料Hoechst 33258是一种与DNA结合的荧光染料，能对这类材料进行更精确的分析。双苯甲酰亚胺能将这些真菌或原生动物有机体的各个阶段都染得很清晰，与均匀的深色背景形成鲜明对比。背景发光水平以及碎屑或非DNA结构的染色都非常低。吞噬细胞内外的有机体被染成相同强度。对单个微生物进行计数，例如在被利什曼原虫严重寄生的巨噬细胞中或在肺孢子菌感染的支气管肺泡灌洗样本中，变得更加简便和精确。空气干燥的细胞悬液、细胞离心涂片、压印涂片或冷冻切片显微照片都显示了双苯甲酰亚胺染色相对于Diff - Quik染色的优势。双苯甲酰亚胺染色可能是分析感染多种微生物的材料的一种有价值的辅助技术。
