title: A simple procedure to improve the surface passivation for single molecule fluorescence studies.
abstract: The single-molecule fluorescence technique is becoming a general and mature tool to probe interactions and dynamics of biomolecules with ultra high precision and accuracy. However, nonspecific adsorption of biomolecules to the flow cells remains a major experimental riddle for the study of many complex biological systems, especially those exhibiting low binding affinity and presenting with weakly populated intermediates. Many novel surface passivation methods have been introduced to reduce nonspecific interactions. Here, we present an effective and inexpensive method to significantly reduce nonspecific binding of biomolecules in conventional poly (ethylene glycol) (PEG)-based surface passivation protocols, without additional exogenous effects. In particular, we propose a simple 10 min Tween-20 treatment for the PEG passivated surface, which could further increase the hydrophilicity of the surface and thus promote passivation efficacy by about 5 to 10 times. We anticipate that this new procedure will find broad practical applications and extend the current reaches of single-molecule fluorescence studies.
authors.affiliation: Collaborative Innovation Center of Advanced Microstructures, National Laboratory of Solid State Microstructure and Department of Physics, Nanjing University, 22 Hankou Road, Nanjing, Jiangsu 210093, People's Republic of China.
authors.forename: Hai
authors.initials: H
authors.lastname: Pan
authors.forename: Yifan
authors.initials: Y
authors.lastname: Xia
authors.forename: Meng
authors.initials: M
authors.lastname: Qin
authors.forename: Yi
authors.initials: Y
authors.lastname: Cao
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
mesh_terms: D000327:Adsorption; D031541:Fluorescence Resonance Energy Transfer; D057927:Hydrophobic and Hydrophilic Interactions; D036103:Nanotechnology / Q000379:methods*; D011092:Polyethylene Glycols / Q000737:chemistry* / Q000378:metabolism; D013499:Surface Properties
chemical_list: D011092:Polyethylene Glycols