title: Analysis of affinity maps of membrane proteins on individual human embryonic stem cells.
abstract: The heterogeneity found in many cell types has greatly inspired research in single-cell gene and protein profiling for discovering the origin of heterogeneity and its role in cell fate decisions. Among the existing techniques to probe heterogeneity, atomic force microscopy (AFM) utilizes an antibody/ligand-modified tip to explore the distribution of a target membrane protein on individual cells in their native environment. In this paper, we establish a practical model to analyze the data systematically, and attempt the quantification of membrane protein abundance on single cells by taking account issues, such as the level of nonspecific interaction, the probe resolution, and the reproducibility of detecting protein distribution. We demonstrated the application in examining the heterogeneous distribution and the local protein abundance of TRA-1-81 antigen on human embryonic stem (hES) cells at the subcellular level. Heterogeneity in TRA-1-81 expression was also detected at the single cell level, suggesting the presence of subpopulation cells within an undifferentiated hES cell colony. The method provides a platform to unveiling the correlation between heterogeneity of membrane proteins and cell development in a complex cell community.
authors.affiliation: Department of Biological, Chemical and Physical Sciences, Illinois Institute of Technology, Chicago, Illinois 60616, USA.
authors.forename: Zhaoxia
authors.initials: Z
authors.lastname: Li
authors.forename: Dengli
authors.initials: D
authors.lastname: Qiu
authors.forename: Kangmin
authors.initials: K
authors.lastname: Xu
authors.forename: Indumathi
authors.initials: I
authors.lastname: Sridharan
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Qian
authors.forename: Rong
authors.initials: R
authors.lastname: Wang
mesh_terms: D000954:Antigens, Surface / Q000737:chemistry*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000737:chemistry*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D013499:Surface Properties
chemical_list: D000954:Antigens, Surface; D008565:Membrane Proteins; C519803:TRA-1-81 antigen, human