Willführ K U, Hirschfeld U, Westermann J, Pabst R
Centre of Anatomy, Medical School of Hannover, F.R.G.
J Immunol Methods. 1990 Jul 3;130(2):201-7. doi: 10.1016/0022-1759(90)90049-2.
The in vitro lymphocyte binding assay (HEV assay) has proved to be a useful approach for examining the first step of lymphocyte migration, i.e., homing to organs containing high endothelial venules (HEVs). Since fluorescence-labelled standard lymphocytes are usually included in each assay to account for day-to-day variations, HEV preparations have to be evaluated by fluorescence microscopy. Thus no counterstaining can be performed and HEVs without adherent lymphocytes cannot easily be recognized. Because the preparations are not suitable for storage they must be evaluated within a short time. In this study an improved technique is described which permits HEV preparations made with fluorescence-labelled standard lymphocytes to be evaluated by light microscopy in counterstained sections. The phenotypes of the sample lymphocytes can be determined by staining for surface antigens on the same slides and the preparations obtained are permanent.
体外淋巴细胞结合试验（高内皮微静脉试验）已被证明是检测淋巴细胞迁移第一步，即归巢至含有高内皮微静脉（HEV）的器官的一种有用方法。由于每次试验通常都包含荧光标记的标准淋巴细胞以解释日常变化，因此HEV制剂必须通过荧光显微镜进行评估。因此，无法进行复染，并且没有粘附淋巴细胞的HEV不易识别。由于制剂不适合储存，因此必须在短时间内进行评估。在本研究中，描述了一种改进的技术，该技术允许在复染切片中通过光学显微镜对用荧光标记的标准淋巴细胞制备的HEV制剂进行评估。样品淋巴细胞的表型可以通过在同一张载玻片上对表面抗原进行染色来确定，并且所获得的制剂是永久性的。
