Label-free detection of surface markers on stem cells by oblique-incidence reflectivity difference microscopy

Kai Yin Lo, Yung Shin Sun, James P. Landry, Xiangdong Zhu, Wenbin Deng

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)381-387
Number of pages7
JournalBioTechniques
Volume50
Issue number6
DOIs
StatePublished - Jun 2011

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CD15 Antigens
Stem cells
Labels
Microscopy
Microscopic examination
Stem Cells
Incidence
Antibodies
Microscopes
Cells
Antibody Specificity
Fluorescence microscopy
Fluorescence Microscopy
Glass
Monitoring

Keywords

  • Cell microarray
  • Label-free
  • Oblique-incidence reflectivity difference (OI-RD)
  • Stem cell

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biotechnology

Cite this

Label-free detection of surface markers on stem cells by oblique-incidence reflectivity difference microscopy. / Lo, Kai Yin; Sun, Yung Shin; Landry, James P.; Zhu, Xiangdong; Deng, Wenbin.

In: BioTechniques, Vol. 50, No. 6, 06.2011, p. 381-387.

Research output: Contribution to journalArticle

Lo, Kai Yin ; Sun, Yung Shin ; Landry, James P. ; Zhu, Xiangdong ; Deng, Wenbin. / Label-free detection of surface markers on stem cells by oblique-incidence reflectivity difference microscopy. In: BioTechniques. 2011 ; Vol. 50, No. 6. pp. 381-387.
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