Conserved linear dynamics of single-molecule Brownian motion

Serag, Maged F.; Habuchi, Satoshi

doi:10.1038/ncomms15675

Cited by 15 publications

(16 citation statements)

References 40 publications

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“…Single-molecule tracking analysis of the PSGL-1 molecules moving along the tethers and slings of KG1a cells rolling over E-selectin and localized on the microvilli of control KG1a cells were conducted by generating single-molecule diffusion trajectories using a versatile tracking algorithm 47 , 48 . The algorithm uses a mixture-model fitting algorithm to localize and track multiple particles in the same field of view.…”

Section: Methodsmentioning

confidence: 99%

Single-molecule imaging and microfluidic platform reveal molecular mechanisms of leukemic cell rolling

et al. 2021

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Hematopoietic stem/progenitor cell (HSPC) and leukemic cell homing is an important biological phenomenon that occurs through key interactions between adhesion molecules. Tethering and rolling of the cells on endothelium, the crucial initial step of the adhesion cascade, is mediated by interactions between selectins expressed on endothelium to their ligands expressed on HSPCs/leukemic cells in flow. Although multiple factors that affect the rolling behavior of the cells have been identified, molecular mechanisms that enable the essential slow and stable cell rolling remain elusive. Here, using a microfluidics-based single-molecule live cell fluorescence imaging, we reveal that unique spatiotemporal dynamics of selectin ligands on the membrane tethers and slings, which are distinct from that on the cell body, play an essential role in the rolling of the cell. Our results suggest that the spatial confinement of the selectin ligands to the tethers and slings together with the rapid scanning of a large area by the selectin ligands, increases the efficiency of selectin-ligand interactions during cell rolling, resulting in slow and stable rolling of the cell on the selectins. Our findings provide novel insights and contribute significantly to the molecular-level understanding of the initial and essential step of the homing process.

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Section: Methodsmentioning

confidence: 99%

Single-molecule imaging and microfluidic platform reveal molecular mechanisms of leukemic cell rolling

et al. 2021

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“…30,31 The data were analyzed as described elsewhere. 32 Sucrose Gradient Sedimentation Assay. The sucrose gradient was prepared by layering less dense sucrose solutions upon one another to achieve a gradient of 5-20% sucrose concentration.…”

Section: Methodsmentioning

confidence: 99%

Geometry-Based Self-Assembly of Histone–DNA Nanostructures at Single-Nucleotide Resolution

et al. 2019

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Histones are basic protein monomers capable of interacting with DNA, providing the mechanism of DNA compaction inside the cell nucleus. The well-ordered assembly process of histone and DNA is a potential candidate as the approach for building DNA-protein nanostructures. Here, utilizing the sequence-independent histone-DNA interaction, we present an approach to selfassemble histones and single-stranded DNA (ssDNA) to form well-defined histone-DNA (sHD) nanoparticles and their multidimensional cross-linked complexes (cHD). By using various molecular biology and microscopy techniques, we elucidate the structure of these complexes, and we show that they are formed at carefully controlled conditions of temperature, ionic-strength, concentration and incubation time. We also demonstrate using a set of ssDNA molecular rulers and a geometric accommodation model that the assembly of sHD and cHD particles proceed with precise geometry so that the number of ssDNA in these particles can be programmed by the length of ssDNA. We further show that the formation of cHD amplifies the effect of the length of ssDNA on the self-assembly, allowing for distinguishing ssDNA of different lengths at single nucleotide resolution. We envision that our geometry-directed approach of self-assembling histone-DNA nanostructures and the fundamental insights can serve as a structural platform to advance building precisely-ordered DNA-protein nanostructures.

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“…The imaging solution was prepared immediately before the imaging experiments. The imaging experiments were conducted on a custom-built wide-field illumination fluorescence microscope on an inverted optical microscope platform (IX71, Olympus) ( 50 , 51 ). A continuous-wave diode laser operating at 638 nm (60 mW; MLD, Cobolt) was introduced into the microscope from its backside port through an achromatic convex lens ( f = 300 mm; Thorlabs) that focused the beam at the back aperture of the objective lens [100×, NA = 1.49; UAPON 100XOTIRF, Olympus].…”

Section: Methodsmentioning

confidence: 99%

Microfluidics-based super-resolution microscopy enables nanoscopic characterization of blood stem cell rolling

et al. 2018

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Conserved linear dynamics of single-molecule Brownian motion

Cited by 15 publications

References 40 publications

Single-molecule imaging and microfluidic platform reveal molecular mechanisms of leukemic cell rolling

Single-molecule imaging and microfluidic platform reveal molecular mechanisms of leukemic cell rolling

Geometry-Based Self-Assembly of Histone–DNA Nanostructures at Single-Nucleotide Resolution

Microfluidics-based super-resolution microscopy enables nanoscopic characterization of blood stem cell rolling

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