Howard R J, Battye F L
Parasitology. 1979 Jun;78(3):263-70. doi: 10.1017/s0031182000051131.
A cell-sorting method is described for the analysis and separation of red blood cells in Plasmodium berghei-infected mouse blood based on their DNA content. This method involves a selective uptake of the bis-benzimidazole dye 33258 Hoechst, a DNA-binding dye, by red blood cells containing parasites. Infected blood is incubated at 37 degrees C with the dye then washed at 4 degrees C to remove unbound dye. Uninfected cells are then non-fluorescent at the characteristic wavelengths for 33258 Hoechst excitation and emission, whereas parasitized cells display fluorescence intensities in approximately direct proportion tothe number of parasite nuclei (i.e. amount of parasite DNA) within the cell and can be sorted accordingly. Providing cells were incubated in a complex nutrient medium during dye uptake at 37 degrees C, the sorted parasite-infected cells produced lethal P. berghei infections when injected into BALB/c mice. The dye-labelling technique is simple and sufficient red blood cells at various stages of infection can be collected for biochemical or immunochemical studies by cell sorting.
描述了一种基于DNA含量分析和分离感染伯氏疟原虫的小鼠血液中红细胞的细胞分选方法。该方法涉及含有寄生虫的红细胞对双苯并咪唑染料33258 Hoechst（一种DNA结合染料）的选择性摄取。将感染的血液与染料在37℃下孵育，然后在4℃下洗涤以去除未结合的染料。未感染的细胞在33258 Hoechst激发和发射的特征波长下无荧光，而被寄生的细胞显示出的荧光强度与细胞内寄生虫核的数量（即寄生虫DNA的量）大致成正比，并可据此进行分选。如果在37℃摄取染料期间将细胞在复杂的营养培养基中孵育，分选的感染寄生虫的细胞注射到BALB/c小鼠中会产生致命的伯氏疟原虫感染。这种染料标记技术简单，通过细胞分选可以收集处于不同感染阶段的足够数量的红细胞用于生化或免疫化学研究。
