A simple procedure to improve the surface passivation for single molecule fluorescence studies

作者：Pan, Hai^*; Xia, Yifan; Qin, Meng; Cao, Yi; Wang, Wei

来源：Physical Biology, 2015, 12(4): 045006.

DOI：10.1088/1478-3975/12/4/045006

• surface passivation
• Tween-20
• single molecule fluorescence
• FRET

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.

• 出版日期2015-7
• 单位固体微结构物理国家重点实验室; 南京大学