Academic Journal
Rapid and improved surface passivation method for Single-Molecule experiments.
| Τίτλος: | Rapid and improved surface passivation method for Single-Molecule experiments. |
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| Συγγραφείς: | Gonneville AN; Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN 37996, USA., Ward AE; Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN 37996, USA., Naidoo NR; Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN 37996, USA., Barrera FN; Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN 37996, USA., Lamichhane R; Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN 37996, USA. Electronic address: rajan@utk.edu. |
| Πηγή: | Methods (San Diego, Calif.) [Methods] 2026 Mar; Vol. 247, pp. 95-106. Date of Electronic Publication: 2026 Jan 07. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Academic Press Country of Publication: United States NLM ID: 9426302 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1095-9130 (Electronic) Linking ISSN: 10462023 NLM ISO Abbreviation: Methods Subsets: MEDLINE |
| Imprint Name(s): | Publication: Duluth, MN : Academic Press Original Publication: San Diego : Academic Press, c1990- |
| Ιατρικοί όροι (MeSH): | Polyethylene Glycols*/chemistry , Fluorescence Resonance Energy Transfer*/methods , Single Molecule Imaging*/methods, Silanes/chemistry ; Fluorescent Dyes/chemistry ; Microscopy, Fluorescence/methods ; Photobleaching ; Surface Properties |
| Περίληψη: | Single-molecule fluorescence experiments are a powerful tool for studying biomolecular interactions, including protein dynamics and oligomerization, protein-protein interactions, and protein-nucleic acid interactions. Biomolecules are commonly immobilized on the microscope surface to extend the observation time. However, non-specific interactions between biomolecules and the surface present a major challenge. The first critical step in these experiments is preparing the surface using polyethylene glycol (PEG) coated slides, which facilitate biomolecule immobilization while minimizing non-specific interactions. The surface treatment typically uses PEG-SVA (Succinimidyl Valerate) coated slides, and the protocol for the treatment is lengthy and time-consuming. To overcome this issue, we have developed a process that uses PEG-Silane to improve efficiency while maintaining reproducibility. Here, we present a one-step, rapid PEGylation methodology that can be completed in minutes rather than hours. We demonstrate its validity and feasibility through single-molecule fluorescence resonance energy transfer (smFRET) and single-molecule photobleaching experiments across various biological samples. (Copyright © 2026 The Authors. Published by Elsevier Inc. All rights reserved.) |
| Competing Interests: | Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. |
| Grant Information: | R35 GM142946 United States GM NIGMS NIH HHS |
| Contributed Indexing: | Keywords: FRET; PEG-Silane; PEGylation; Photobleaching; Single-molecule; TIRF microscopy |
| Substance Nomenclature: | 3WJQ0SDW1A (Polyethylene Glycols) 0 (Silanes) 0 (Fluorescent Dyes) |
| Entry Date(s): | Date Created: 20260109 Date Completed: 20260625 Latest Revision: 20260625 |
| Update Code: | 20260626 |
| PubMed Central ID: | PMC12933383 |
| DOI: | 10.1016/j.ymeth.2026.01.003 |
| PMID: | 41512980 |
| Βάση Δεδομένων: | MEDLINE |
| ISSN: | 1095-9130 |
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| DOI: | 10.1016/j.ymeth.2026.01.003 |