ArticleJournal of visualized experiments : JoVE2014
Surface passivation for single-molecule protein studies.
Article in Journal of visualized experiments : JoVE, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 126 papers.
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Who cites it
126 citing papers in PubMed.
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- Tau Aggregate Imaging and Transcriptomics of Alzheimer's Disease Brain at Different Stages of Disease.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Direct single-molecule visualization of Hsp90-mediated relief of an Hsp70-folding block.Science advances · 2026Article
- ATP-driven conformational dynamics reveal hidden intermediates in a heterodimeric ABC transporter.eLife · 2026Article
- Chromosome topology gates productive RecA homology search.bioRxiv : the preprint server for biology · 2026Article
- smDeepFLUOR: single-molecule deep learning fluorescence classification.Nature communications · 2026Article
- Chaperones shape the conformational landscape of 26S-proteasome-base assembly for allosteric ATPase motor activation.bioRxiv : the preprint server for biology · 2026Article
- Reconstitution of Active Plant HBio-protocol · 2026Article
- Mechanisms of human mitochondrial leaderless mRNA translation initiation.Nature communications · 2026Article
- Single-molecule dynamics reveal ATP binding alone powers substrate translocation by an ABC transporter.Nature communications · 2026Article
- Functional characterization of dynamic nascent RNA folding ensembles in real time.Science advances · 2026Article
- Rapid and improved surface passivation method for Single-Molecule experiments.Methods (San Diego, Calif.) · 2026Article
- Ultrafast and specific miRNA quantification via single-molecule fluorescence quenching kinetics.Communications biology · 2026Article
- Surface-Tethering Enhances Precision in Measuring Diffusion Within 3D Protein Condensates.Journal of molecular biology · 2026Article
- DNA Hanger: Surface-Minimized Single-Molecule Immunoassay Platform.Small (Weinheim an der Bergstrasse, Germany) · 2025Article
- Reconstitution of circadian clock in synthetic cells reveals principles of timekeeping.Nature communications · 2025Article
- Surface-Tethering Enhances Precision in Measuring Diffusion Within 3D Protein Condensates.bioRxiv : the preprint server for biology · 2025Article
- Inhibition of p38-MK2 pathway enhances the efficacy of microtubule inhibitors in breast cancer cells.eLife · 2025Article
- Inhibition of p38-MK2 pathway enhances the efficacy of microtubule inhibitors in breast cancer cells.bioRxiv : the preprint server for biology · 2025Article
66 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Single-molecule fluorescence spectroscopy has proven to be instrumental in understanding a wide range of biological phenomena at the nanoscale. Important examples of what this technique can yield to biological sciences are the mechanistic insights on protein-protein and protein-nucleic acid interactions. When interactions of proteins are probed at the single-molecule level, the proteins or their substrates are often immobilized on a glass surface, which allows for a long-term observation. This immobilization scheme may introduce unwanted surface artifacts. Therefore, it is essential to passivate the glass surface to make it inert. Surface coating using polyethylene glycol (PEG) stands out for its high performance in preventing proteins from non-specifically interacting with a glass surface. However, the polymer coating procedure is difficult, due to the complication arising from a series of surface treatments and the stringent requirement that a surface needs to be free of any fluorescent molecules at the end of the procedure. Here, we provide a robust protocol with step-by-step instructions. It covers surface cleaning including piranha etching, surface functionalization with amine groups, and finally PEG coating. To obtain a high density of a PEG layer, we introduce a new strategy of treating the surface with PEG molecules over two rounds, which remarkably improves the quality of passivation. We provide representative results as well as practical advice for each critical step so that anyone can achieve the high quality surface passivation.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.