Immunogold-Silver Staining Method for Lymphocyte Cell Surface Antigens with Light, Transmission Electron, and Scanning Electron Microscopy
Immunogold-Silver Staining Method for Lymphocyte Cell Surface Antigens with Light, Transmission Electron, and Scanning Electron Microscopy
复制标题
淋巴细胞表面抗原的光、透射电子和扫描电镜免疫金银染色方法
DOI：
10.1179/his.1993.16.3.217
复制
发表时间：
1993
期刊：
影响因子：
--
通讯作者：
L. Maxwell
中科院分区：
文献类型：
--
作者：
Y. Otsuki;L. Maxwell
摘要采用免疫金银染色(IGSS)法结合光镜、透射电子显微镜和扫描电子显微镜(分别为光镜、透射电子显微镜和扫描电子显微镜)检测淋巴细胞表面抗原。两种不同大小的胶体金粒子(5NRN和15 nm)作为标记，IntenSEII试剂盒作为金粒子的物理显色剂。在光镜和电子显微镜下，银增强的金粒子在细胞表面清晰地显示为黑点，在配备二次电子和背散射电子(SE/BE)混合信号的扫描电子显微镜中，银增强的金粒子清晰地显示为白点。在配备SE/BE信号的低倍光镜或扫描电子显微镜下，在保存完好的表面含有复合体的单抗(MAb)阳性细胞很容易与其他淋巴样细胞区分开来。因此，IGSS方法在细胞悬液中淋巴细胞亚群百分比的定性筛选方面具有很大的优势。然而，IGSS方法不能用于抗原的半定量研究。
Abstract The immunogold-silver staining (IGSS) method combined with light, transmission electron, and scanning electron microscopy (LM, TEM and SEM, respectively) was used for detecting lymphocyte surface antigens. Two different sizes of colloidal gold particles (5 nrn and 15 nm) were applied as markers and IntenSEII kit as a physical developer for gold particles.The silver enhanced gold particles were clearly observed on cell surfaces as black dots in LM and TEM and as white dots in SEM equipped with a mixed signal of secondary electron and back-scattered electron (SE/BE) signals. Monoclonal antibody (MAb)-positive cells possessing the complexes on their well preserved surfaces were easily identified among other lymphoid cells at low magnifications of LM or SEM equipped with SE/BE signals. Thus, the IGSS method has a great advantage for a qualitative screening such as the percentage of lymphocyte subsets in a cell suspension. However, the IGSS method was inadequate for semiquantitative study with antigen...