| RRC ID |
38544
|
| 著者 |
Lo KY, Sun YS, Landry JP, Zhu X, Deng W.
|
| タイトル |
Label-free detection of surface markers on stem cells by oblique-incidence reflectivity difference microscopy.
|
| ジャーナル |
Biotechniques
|
| Abstract |
Conventional fluorescence microscopy is routinely used to detect cell surface markers through fluorophore-conjugated antibodies. However, fluorophore-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 microarray and reacted the cell microarray 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.
|
| 巻・号 |
50(6)
|
| ページ |
381-8
|
| 公開日 |
2011-6-1
|
| DOI |
10.2144/000113670
|
| PII |
000113670
|
| PMID |
21781038
|
| PMC |
PMC3271734
|
| MeSH |
Animals
Antibodies / metabolism
Biomarkers / chemistry
Cell Differentiation
Cells, Immobilized / cytology
Cells, Immobilized / metabolism*
Cytological Techniques
Lewis X Antigen / chemistry*
Lewis X Antigen / metabolism
Mice
Microscopy / methods*
Microscopy, Fluorescence
Protein Binding
Stem Cells / chemistry*
Stem Cells / cytology
Stem Cells / metabolism
Tissue Array Analysis / methods*
|
| IF |
1.541
|
| 引用数 |
8
|
|
WOS 分野
|
BIOCHEMICAL RESEARCH METHODS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
mouse induced pluripotent stem (miPS) cells |
