2022
DOI: 10.1021/acsami.2c16647
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Surface Passivation with a Perfluoroalkane Brush Improves the Precision of Single-Molecule Measurements

Abstract: Single-molecule imaging is invaluable for investigating the heterogeneous behavior and interactions of biological molecules. However, an impediment to precise sampling of single molecules is the irreversible adsorption of components onto the surfaces of cover glasses. This causes continuous changes in the concentrations of different molecules dissolved or suspended in the aqueous phase from the moment a sample is dispensed, which will shift, over time, the position of chemical equilibria between monomeric and … Show more

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Cited by 15 publications
(17 citation statements)
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“…We recently demonstrated the suitability of perfluoroalkane brushes to monitor adsorption and desorption of single proteins over a prolonged duration via iSCAT microscopy, and their potential to study thermodynamic equilibria between monomeric and multimeric protein components. 8 This work expands on our previous results and presents an optimised protocol for the formation of perfluorinated SAMs on cover glasses, which have been tested with a commercial protein mixture. We believe these results are just the first example of many experimental possibilities that can be unlocked using perfluorinated monolayers for single-molecule studies.…”
Section: Introductionmentioning
confidence: 66%
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“…We recently demonstrated the suitability of perfluoroalkane brushes to monitor adsorption and desorption of single proteins over a prolonged duration via iSCAT microscopy, and their potential to study thermodynamic equilibria between monomeric and multimeric protein components. 8 This work expands on our previous results and presents an optimised protocol for the formation of perfluorinated SAMs on cover glasses, which have been tested with a commercial protein mixture. We believe these results are just the first example of many experimental possibilities that can be unlocked using perfluorinated monolayers for single-molecule studies.…”
Section: Introductionmentioning
confidence: 66%
“…Protein 2 results in the most intense peak after 1 minute, which indicates it presents the largest concentration, followed by protein 1 and protein 3. Using glass substrates, these peaks tend to be less intense over time due to the irreversible adsorption of proteins onto the substrate, 8 which limits the time span of the measurements. However, since the perfluorinated SAM prevents irreversible adsorption events, we were able take measurements for several minutes.…”
Section: Iscatmentioning
confidence: 99%
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“…5 The lack of intramolecular interactions between fluorous moieties and other species makes fluorous layers good anti-fouling platforms. 6 Furthermore, fluorous-fluorous interactions are non-covalent and highly specific, and can be employed as orthogonal recognition modes 7 or for reversible molecular patterning. 8 We aim to expand these functionalities to build new reversible systems.…”
Section: Introductionmentioning
confidence: 99%
“…Recent developments in ultrasensitive light microscopy 1,2 have enabled the quantification of biomolecular mass, charge and size at the single molecule level and in solution [3][4][5][6][7][8] . In mass photometry (MP) light scattered from a protein when it binds to or moves along a glass coverslip in solution is detected together with partially reflected light from the glass-water interface (Fig.…”
Section: Introductionmentioning
confidence: 99%