A new platform for single molecule measurements using the fluorous effect
A new platform for single molecule measurements using the fluorous effect
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DOI：
10.1117/12.2647953
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发表时间：
2023-03
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通讯作者：
M. Santana Vega;Carlos J Bueno-Alejo;Andrea Taladriz Sender;A. Chaplin;Chloe Farrow;Alexander Axer;G. Burley;Cyril Dominguez;Hesna Kara;Vasileious Paschalis;Sumera Tubasum;I. Eperon;A. J. Hudson;A. Clark
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作者：
M. Santana Vega;Carlos J Bueno-Alejo;Andrea Taladriz Sender;A. Chaplin;Chloe Farrow;Alexander Axer;G. Burley;Cyril Dominguez;Hesna Kara;Vasileious Paschalis;Sumera Tubasum;I. Eperon;A. J. Hudson;A. Clark
生物分子在成像基底上的不可逆吸附是扩大单分子技术应用的障碍。传统的聚乙二醇 (PEG) 表面仅在低浓度的分析物下有效，且其结构阻碍了其用于干涉散射 (iSCAT) 显微镜。我们提出了一个新平台，由于全氟化合物的全疏性（也称为氟效应），该平台实际上消除了非特异性结合。在这里，我们通过蛋白质混合物的 iSCAT 测量在单分子水平上展示了这些底物的防污特性。我们相信，通过 iSCAT 显微镜和其他单分子技术，这些新颖的工程基质在研究需要大分析物浓度的生物机械过程方面显示出巨大的前景，而其他钝化方法在这些过程中无效。
Irreversible adsorption of biomolecules onto imaging substrates is an impediment to expand the applications of single molecule techniques. Traditional polyethylene glycol (PEG) surfaces are only effective at low concentrations of analytes and their structure prevents their use for interferometric scattering (iSCAT) microscopy. We propose a new platform that virtually eliminates non-specific binding thanks to the omniphobicity of perfluorinated compounds, also known as the fluorous effect. Here, we showcase the anti-fouling properties of these substrates at a single molecule level through iSCAT measurements of a protein mixture. We believe these novel engineered substrates show great promise to study biomachinery processes requiring large analyte concentrations, where other passivation methods are not effective, through iSCAT microscopy and other single molecule techniques.