Vesicle sizing

Hallett, F. R.; Watton, J.; Krygsman, Peter H.

doi:10.1016/s0006-3495(91)82229-9

Cited by 102 publications

(44 citation statements)

References 27 publications

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“…OMVs were measured by dynamic light scattering (DLS) using a Zetasizer Nano ZS (Malvern Instruments) for detecting the size distribution, which was reflected in the polydispersity index (PdI), ranging between 0.0 (monodispersed) and 1.0 (entirely heterodispersed)5152. Next, the OMVs were observed and imaged using a transmission electron microscope (Hitachi) to analyze their morphology after fixing with 2.5% cold glutaraldehyde in 0.2 M sodium cacodylate buffer (pH 7.4) for 2 h at 4 °C and post-fixing with 1% osmium tetroxide in 0.1 M sodium cacodylate buffer (pH 7.4) for 1 h at 4 °C.…”

Section: Methodsmentioning

confidence: 99%

Employing Escherichia coli-derived outer membrane vesicles as an antigen delivery platform elicits protective immunity against Acinetobacter baumannii infection

Huang

Wang

Yao

et al. 2016

Sci Rep

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Outer membrane vesicles (OMVs) have proven to be highly immunogenic and induced an immune response against bacterial infection in human clinics and animal models. We sought to investigate whether engineered OMVs can be a feasible antigen-delivery platform for efficiently inducing specific antibody responses. In this study, Omp22 (an outer membrane protein of A. baumannii) was displayed on E. coli DH5α-derived OMVs (Omp22-OMVs) using recombinant gene technology. The morphological features of Omp22-OMVs were similar to those of wild-type OMVs (wtOMVs). Immunization with Omp22-OMVs induced high titers of Omp22-specific antibodies. In a murine sepsis model, Omp22-OMV immunization significantly protected mice from lethal challenge with a clinically isolated A. baumannii strain, which was evidenced by the increased survival rate of the mice, the reduced bacterial burdens in the lung, spleen, liver, kidney, and blood, and the suppressed serum levels of inflammatory cytokines. In vitro opsonophagocytosis assays showed that antiserum collected from Omp22-OMV-immunized mice had bactericidal activity against clinical isolates, which was partly specific antibody-dependent. These results strongly indicated that engineered OMVs could display a whole heterologous protein (~22 kDa) on the surface and effectively induce specific antibody responses, and thus OMVs have the potential to be a feasible vaccine platform.

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Section: Methodsmentioning

confidence: 99%

Employing Escherichia coli-derived outer membrane vesicles as an antigen delivery platform elicits protective immunity against Acinetobacter baumannii infection

Huang

Wang

Yao

et al. 2016

Sci Rep

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“…This can be accomplished by using a manual syringe apparatus or using external pressure from a N 2 tank. The size and polydispersity of SUVs can be measured using dynamic light scattering 37 . ■ PAUSE POINT Vesicles can be used without loss of function for at least 2 weeks if stored at 4 °C.…”

Section: Methodsmentioning

confidence: 99%

Using patterned supported lipid membranes to investigate the role of receptor organization in intercellular signaling

et al. 2011

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physical inputs, both internal and external to a cell, can directly alter the spatial organization of cell surface receptors and their associated functions. Here we describe a protocol that combines solid-state nanolithography and supported lipid membrane techniques to trigger and manipulate specific receptors on the surface of living cells and to develop an understanding of the interplay between spatial organization and receptor function. While existing protein-patterning techniques are capable of presenting cells with well-defined clusters of protein, this protocol uniquely allows for the control of the spatial organization of laterally fluid receptor-ligand complex at an intermembrane junction. a combination of immunofluorescence and single-cell microscopy methods and complementary biochemical analyses are used to characterize receptor signaling pathways and cell functions. the protocol requires 2–5 d to complete depending on the parameters to be studied. In principle, this protocol is widely applicable to eukaryotic cells and herein is specifically developed to study the role of physical organization and translocation of the EphA2 receptor tyrosine kinase across a library of model breast cancer cell lines.

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“…The Schulz distribution has been used successfully to describe the size distribution of a variety of colloidal aggregates (Aragò n & Pecora, 1976;Hayter, 1983;Kotlarchyk & Chen, 1983;Bartlett & Ottewill, 1992;Wagner, 2004). As demonstrated by electron microscopy (Hallett, Nickel et al, 1991), light scattering (Hallett, Watton & Krygsman, 1991) and SANS (Pencer, Mills et al, 2005), the size distribution of extruded phospholipid vesicles is well described by a Schulz distribution. Despite the successful use of the Schulz and other similar continuous functions to describe vesicle size distributions, some caution should be exercised to avoid the use of non-physical parameters (e.g.…”

Section: The Laplace Methodsmentioning

confidence: 99%

Method of separated form factors for polydisperse vesicles

et al. 2006

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Use of the Schulz or Gamma distribution in the description of particle sizes facilitates calculation of analytic polydisperse form factors using Laplace transforms, L[f(u)]. Here, the Laplace transform approach is combined with the separated form factor (SFF) approximation [Kiselev et al. (2002). Appl. Phys. A, 74, S1654-S1656] to obtain expressions for form factors, P(q), for polydisperse spherical vesicles with various forms of membrane scattering length density (SLD) profile. The SFF approximation is tested against exact form factors that have been numerically integrated over the size distribution, and is shown to represent the vesicle form factor accurately for typical vesicle sizes and membrane thicknesses. Finally, various model SLD profiles are used with the SFF approximation to fit experimental small-angle neutron scattering (SANS) curves from extruded unilamellar vesicles.

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Vesicle sizing

Cited by 102 publications

References 27 publications

Employing Escherichia coli-derived outer membrane vesicles as an antigen delivery platform elicits protective immunity against Acinetobacter baumannii infection

Employing Escherichia coli-derived outer membrane vesicles as an antigen delivery platform elicits protective immunity against Acinetobacter baumannii infection

Using patterned supported lipid membranes to investigate the role of receptor organization in intercellular signaling

Method of separated form factors for polydisperse vesicles

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