https://scholars.lib.ntu.edu.tw/handle/123456789/444644
標題: | Label-free detection of surface markers on stem cells by oblique-incidence reflectivity difference microscopy | 作者: | KAI-YIN LO Sun, Y.-S. Landry, J.P. Zhu, X. |
關鍵字: | Cell microarray | Label-free | Oblique-incidence reflectivity difference (OI-RD) | Stem cell | 公開日期: | 2011 | 出版社: | BIOTECHNIQUES OFFICE | 卷: | 50 | 期: | 6 | 起(迄)頁: | 381 | 來源出版物: | BioTechniques | 摘要: | Conventional fluorescence microscopy is routinely used to detect cell surface markers through foreshore-conjugated antibodies. However, fuorophore-conjugation of antibodies alters binding properties such as strength and specificity of the antibody in often uncharacterized ways. Here we present a method using an oblique-incidence reflectivity difference (OI-RD) microscope for label-free, real-time detection of cell surface markers, and apply it to analysis of stage-specific embryonic antigen 1 (SSEA1) on stem cells. Mouse stem cells express SSEA1 on their surfaces, and the level of SSEA1 decreases when the cells start to differentiate. In this study, we immobilized mouse stem cells and non-stem cells (control) on a glass surface as a micro array and reacted the cell micro array with unlabeled SSEA1 antibodies. By monitoring the reaction with an OI-RD microscope in real time, we confirmed that the SSEA1 antibodies bind only to the surface of the stem cells and not to the surface of non-stem cells. From the binding curves, we determined the equilibrium dissociation constant (Kd) of the antibody with the SSEA1 markers on the stem cell surface. Thus, the OI-RD microscope can be used to detect binding affinities between cell surface markers and unlabeled antibodies bound to the cells; this information could be useful for determination of stem cell stages. |
URI: | https://scholars.lib.ntu.edu.tw/handle/123456789/444644 | ISSN: | 0736-6205 | DOI: | 10.2144/000113670 23124205 |
顯示於: | 農業化學系 |
在 IR 系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。