Resistive-pulse sensing with biological or solid-state nanopores and nanopipettes has been widely employed in detecting single molecules and nanoparticles. The analytical signal in such experiments is the change in ionic current caused by the molecule/particle translocation through the pipet orifice. This paper describes a new version of the resistive-pulse technique based on the use of carbon nanopipettes (CNP). The measured current is produced by electrochemical oxidation/reduction of redox molecules at the carbon surface and responds to the particle translocation. In addition to counting single entities, this technique enables qualitative and quantitative analysis of the electroactive material they contain. Using liposomes as a model system, we demonstrate the capacity of CNPs for (1) conventional resistive-pulse sensing of single liposomes, (2) electrochemical resistive-pulse sensing, and (3) electrochemical identification and quantitation of redox species (e.g., ferrocyanide, dopamine, and nitrite) contained in a single liposome. The small physical size of a CNP suggests the possibility of single-entity measurements in biological systems.
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Int J Mol Sci
December 2024
Nantes Université, Oniris, CHU Nantes, Inserm, Regenerative Medicine and Skeleton, RMeS, UMR 1229, F-44000 Nantes, France.
Inflammation significantly influences cellular communication in the oral environment, impacting tissue repair and regeneration. This study explores the role of small extracellular vesicles (sEVs) derived from lipopolysaccharide (LPS)-treated stem cells from the apical papilla (SCAP) in modulating macrophage polarization and osteoblast differentiation. SCAPs were treated with LPS for 24 h, and sEVs from untreated (SCAP-sEVs) and LPS-treated SCAP (LPS-SCAP-sEVs) were isolated via ultracentrifugation and characterized using transmission electron microscopy, Western blot, and Tunable Resistive Pulse Sensing.
View Article and Find Full Text PDFACS Nano
January 2025
Department of Chemistry, Indiana University, Bloomington, Indiana 47405-7102, United States.
Characterization of individual biological nanoparticles can be significantly improved by coupling complementary analytical methods. Here, we combine resistive-pulse sensing (RPS) with fluorescence lifetime imaging microscopy (FLIM) to differentiate liposomes at the single-particle level. RPS measures the particle volume, shape, and surface-charge density, and FLIM determines the fluorescence lifetime of the fluorophore associated with the lipid membrane.
View Article and Find Full Text PDFAnal Chem
December 2024
Department of Chemistry, The University of Kansas, Lawrence, Kansas 66045, United States.
We are developing a unique protein identification method that consists of generating peptides proteolytically from a single protein molecule (i.e., peptide fingerprints) with peptide detection and identification carried out using nanoscale electrochromatography and label-free resistive pulse sensing (RPS).
View Article and Find Full Text PDFJ Extracell Vesicles
October 2024
Université Paris-Cité, PARCC, INSERM, Paris, France.
Virology
December 2024
Molecular and Cellular Biochemistry Department, Indiana University, Bloomington, IN, USA. Electronic address:
Human Papillomavirus serotype 16 (HPV16) capsid protein (L1) pentamers canonically assemble into T = 7 icosahedral capsids. Such virus-like particles are the basis of the HPV vaccine. We examined assembly of L1 pentamers in response to pH, mild oxidants, and ionic strength and found a mixture of closed, roughly spherical structures from ∼20 to ∼70 nm in diameter, indicating the presence of many kinetically accessible energy minima.
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