A new platform for single molecule measurements using the fluorous effect

Marina Santana Vega; Carlos J. Bueno-Alejo; Andrea Taladriz Sender; Amanda K. Chaplin; Chloe Farrow; Alexander Axer; Glenn A. Burley; Cyril Dominguez; Hesna Kara; Vasileious Paschalis; Sumera Tubasum; Ian C. Eperon; Andrew J. Hudson; Alasdair W. Clark

doi:10.1117/12.2647953

15 March 2023 A new platform for single molecule measurements using the fluorous effect

Marina Santana Vega, Carlos J. Bueno-Alejo, Andrea Taladriz Sender, Amanda K. Chaplin, Chloe Farrow, Alexander Axer, Glenn A. Burley, Cyril Dominguez, Hesna Kara, Vasileious Paschalis, Sumera Tubasum, Ian C. Eperon, Andrew J. Hudson, Alasdair W. Clark

Author Affiliations +

Proceedings Volume 12386, Single Molecule Spectroscopy and Superresolution Imaging XVI; 123860A (2023) https://doi.org/10.1117/12.2647953
Event: SPIE BiOS, 2023, San Francisco, California, United States

Abstract

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.

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Citation Download Citation

Marina Santana Vega, Carlos J. Bueno-Alejo, Andrea Taladriz Sender, Amanda K. Chaplin, Chloe Farrow, Alexander Axer, Glenn A. Burley, Cyril Dominguez, Hesna Kara, Vasileious Paschalis, Sumera Tubasum, Ian C. Eperon, Andrew J. Hudson, and Alasdair W. Clark "A new platform for single molecule measurements using the fluorous effect", Proc. SPIE 12386, Single Molecule Spectroscopy and Superresolution Imaging XVI, 123860A (15 March 2023); https://doi.org/10.1117/12.2647953

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