Category Ranking
98%
Total Visits
921
Avg Visit Duration
2 minutes
Citations
20
Article Abstract
Interferometric scattering (iSCAT) microscopy enables the label-free observation of biomolecules. Consequently, single-particle imaging and tracking with the iSCAT-based method known as mass photometry (MP) is a growing area of study. However, establishing reliable cover glass passivation and functionalisation methods is crucial to reduce nonspecific binding and prepare surfaces for in vitro single-molecule binding experiments. Existing protocols for fluorescence microscopy can contain strongly scattering or mobile components, which make them impractical for MP-based microscopy. In this study, we characterise several different surface coatings using MP. We present approaches for cover glass passivation using 3-aminopropyltriethoxysilane (APTES) and polyethylene glycol (PEG, 2k) along with functionalisation via a maleimide-thiol linker. These coatings are compatible with water or salt buffers, and show low background scattering; thus, we are able to measure proteins as small as 60 kDa. In this technical note, we offer a surface preparation suitable for in vitro experiments with MP.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1111/jmi.13302 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Surface passivation and functionalisation for mass photometry.
J Microsc
July 2024
Laboratory of Experimental Biophysics (LEB), Institute of Physics and Institute of Bioengineering, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.
Interferometric scattering (iSCAT) microscopy enables the label-free observation of biomolecules. Consequently, single-particle imaging and tracking with the iSCAT-based method known as mass photometry (MP) is a growing area of study. However, establishing reliable cover glass passivation and functionalisation methods is crucial to reduce nonspecific binding and prepare surfaces for in vitro single-molecule binding experiments.
View Article and Find Full Text PDFPrecise determination of molecular adsorption geometries by room temperature non-contact atomic force microscopy.
Commun Chem
January 2024
The University of Leeds, Leeds, United Kingdom.
High resolution force measurements of molecules on surfaces, in non-contact atomic force microscopy, are often only performed at cryogenic temperatures, due to needing a highly stable system, and a passivated probe tip (typically via CO-functionalisation). Here we show a reliable protocol for acquiring three-dimensional force map data over both single organic molecules and assembled islands of molecules, at room temperature. Isolated cobalt phthalocyanine and islands of C are characterised with submolecular resolution, on a passivated silicon substrate (B:Si(111)-[Formula: see text]).
View Article and Find Full Text PDFpH-Activated Dissolvable Polymeric Coatings to Reduce Biofouling on Electrochemical Sensors.
J Funct Biomater
June 2023
School of Chemistry, University of Edinburgh, Joseph Black Building, West Mains Road, Edinburgh EH9 3FJ, UK.
Implantable electrochemical sensors that enable the real-time detection of significant biomarkers offer huge potential for the enhancement and personalisation of therapies; however, biofouling is a key challenge encountered by any implantable system. This is particularly an issue immediately after implantation, when the foreign body response and associated biofouling processes are at their most active in passivating a foreign object. Here, we present the development of a sensor protection and activation strategy against biofouling, based on coatings consisting of a pH-triggered, dissolvable polymer, that covered a functionalised electrode surface.
View Article and Find Full Text PDFPlatinum deposition on functionalised graphene for corrosion resistant oxygen reduction electrodes.
J Mater Chem A Mater
September 2022
Department of Chemistry, MSRH, Imperial College London W12 0BZ UK
Graphene-related materials are promising supports for electrocatalysts due to their stability and high surface area. Their innate surface chemistries can be controlled and tuned functionalisation to improve the stability of both the carbon support and the metal catalyst. Functionalised graphenes were prepared using either aryl diazonium functionalisation or non-destructive chemical reduction, to provide groups adapted for platinum deposition.
View Article and Find Full Text PDFAcid resistant functionalised magnetic nanoparticles for radionuclide and heavy metal adsorption.
J Colloid Interface Sci
February 2022
Department of Materials, Imperial College London, Exhibition Road, London SW7 2AZ, UK; London Centre of Nanotechnology, Imperial College London, Exhibition Road, London SW7 2AZ, UK. Electronic address:
Coating superparamagnetic iron oxide NPs with SiO has been established in order to confer stability in acidic media. Acid stability tests were carried out between pH 1 and pH 7 to determine the effectiveness of the SiO passivating layer to protect the magnetic FeO core. Transmission Electron Microscopy (TEM) and zeta potential measurements have shown that uncoated FeO NPs exhibit rapid agglomeration and dissolution when exposed to acidic media, moving from a zeta potential of - 26 mV to a zeta potential of + 3 mV.
View Article and Find Full Text PDF