Kalland T
J Immunol Methods. 1977;17(3-4):279-83. doi: 10.1016/0022-1759(77)90110-7.
A method is described for rapid and simple visualization of lymphocytes in the rosette assay for human T-lymphocytes. A drop of acridine orange (pH 7.4) is added to the suspension of rosetted cells and allowed to incubate for 1 min on ice. Nucleic acids are then visualized in a fluorescence microscope by excitation at 430-500 micrometer. Simultaneous semi-illumination with light microscopy allows enumeration of total lymphocytes and the number of lymphocytes rosetted with sheep red blood cells. The central lymphocyte is clearly distinguishable even when it is crowded with red cells, and this makes the differentiation between rosettes and non-specific aggregation of cells easy.
描述了一种用于在人T淋巴细胞的玫瑰花结试验中快速简单地观察淋巴细胞的方法。将一滴吖啶橙（pH 7.4）加入到形成玫瑰花结的细胞悬液中，并在冰上孵育1分钟。然后在荧光显微镜下通过430 - 500微米的激发光观察核酸。同时用光学显微镜进行半照明，以便计数总淋巴细胞数以及与绵羊红细胞形成玫瑰花结的淋巴细胞数。即使中央淋巴细胞被红细胞挤满，也能清晰区分，这使得区分玫瑰花结和细胞的非特异性聚集变得容易。
