中性红、吖啶橙及PE标记的LAMP-2抗体在B16F10细胞溶酶体检测中的应用与比较  

Application and comparison of neutral red,acridine orange,or PE labeled LAMP-2 antibodies in the detection of lysosomes in B16F10 cells

作　　者：翟晓峰[1] 施文[1] 李国兴[2] 孙永强[1] 赵文静[2] 钱红燕[2] 李静[2] 陈橼[2] 何向锋[2] 

机构地区：[1]南通大学第二附属医院普外科,226001 [2]南通大学附属肿瘤医院中心实验室,226361

出　　处：《免疫学杂志》2012年第12期1069-1072,共4页Immunological Journal

摘　　要：目的探讨中性红、吖啶橙及PE标记的LAMP-2抗体在小鼠黑色素瘤B16F10细胞溶酶体检测中的应用与价值。方法分别用中性红、吖啶橙和PE标记的LAMP-2抗体标记B16F10细胞溶酶体,通过光学和荧光显微镜进行检测。结果中性红染色法能够在光镜下清晰显示溶酶体在细胞中的分布与数量,并能够反应溶酶体的功能;吖啶橙能够同时将细胞质和溶酶体分别用绿色和红色荧光标示出来,但溶酶体的红色荧光淬灭很快,观测窗口较窄;PE标记的LAMP-2抗体能够将溶酶体在细胞内的位置和数量清晰以红色荧光呈现出来,配合DAPI染色,可以直观显示溶酶体同细胞核的相对位置关系。结论中性红、吖啶橙和PE标记的LAMP-2抗体在溶酶体检测中具有不同的特点和优势,可根据实验需求灵活选用。We aimed to investigate the application and value of neutral red, acridine orange, or PE labeled anti-LAMP-2 antibodies in the detection of lysosomes in B16F10 cells. Firstly, we labeled the lysosomes of B16F10 cells with neutral red, acridine orange, and PE labeled anti-LAMP-2 antibodies respectively and detect the labeled lysosomes by optical and fluorescent microscopy. The results showed that the distribution and numbers of lysosomes in B16F10 cells could be clearly observed in optical microscope through the staining of neutral red, and the cytoplasm and lysosome could be stained in green and red respectively by acridine orange, but the quenching of red fluorescence in lysosome was so fast that the detection window was too narrow. The location and numbers of lysosomes in B16F10 cells could be clearly revealed with red fluorescence after the application of PE- labeled anti-LAMP-2 antibody, and the relative location of lysosomes and nucleus could be presented directly along with DAPI staining. In conclusion, the neutral red, acridine orange and PE labeled LAMP-2 antibody staining have their own advantage and characteristics in the detection of lysosomes. The choice of the effective lysosomes detection method should be based on the aim of study.

关 键 词：溶酶体 中性红 吖啶橙 LAMP-2 荧光染色 

分 类 号：R446.61[医药卫生—诊断学]