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将PHA-L(Phaseolus Vulgaris-Leucoagglutinin,植物血凝素L)导入中枢神经系统特定部位,在可能投射区作免疫组化染色,可取得理想的顺行标记结果。我们用Vector实验室产PHA-抗原,根据微量免疫法免疫家兔,获得了效价较好的兔抗PHA-L抗血清。成年雄性新西兰纯种兔三只,脚掌皮下注入Freund完全佐剂,待触及鼠膝部或腘窝淋巴结肿大成黄豆至蚕豆大小时,将与Freund不完全佐剂混匀成油色水状态的PHA-L抗原0.1ml注入每例动物肿大之淋巴结。一个月后于每例动物背部皮下多点注入抗原。又一个月重复注射。注射后12天耳缘静脉采血,用琼脂双扩法测抗血清效价,三例动物中,有一例动物效价为1∶32—1∶64。第14天颈动脉放血,分离血清。
PHA-L (Phaseolus Vulgaris-Leucoagglutinin) is introduced into specific parts of the central nervous system and immunohistochemical staining in the possible projection area can achieve ideal paraffin labeling. We used Vector lab to produce PHA-antigen and immunized rabbits according to the microimmunization method to obtain the rabbit anti-PHA-L antiserum with better titer. Three adult male New Zealand purebred rabbits were injected subcutaneously with Freund’s complete adjuvant. When the knee or popliteal lymph nodes were swollen into soybeans and broad beans, they were mixed with Freund’s incomplete adjuvant to form a greyish-colored PHA -L antigen 0.1ml into each animal swollen lymph nodes. One month later, each animal was subcutaneously injected with antigens at the back of the animal. Repeat injection another month. Blood was collected from the ear vein 12 days after injection and antiserum titers were determined by agar double-diffusion. In one of the three animals, the titer of the animal was 1:32 to 1:64. The carotid artery was bled on day 14 and the serum was separated.