抗原变异可使得多种致病微生物易于逃避宿主免疫应答。表达在感染红细胞表面的恶性疟原虫红细胞表面蛋白1（PfPMP1）与感染红细胞、内皮细胞、树突状细胞以及胎盘的单个或多个受体作用，在黏附及免疫逃避中起关键的作用。每个单倍体基因组var基因家族编码约60种成员，通过启动转录不同的var基因变异体为抗原变异提供了分子基础。

疟原虫var基因转换速率变化导致抗原变异[英]／Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A

Nanogels and microgels are soft, deformable, and penetrable objects with an internal gel-like structure that is swollen by the dispersing solvent. Their softness and the potential to respond to external stimuli like temperature, pressure, pH, ionic strength, and different analytes make them interesting as soft model systems in fundamental research as well as for a broad range of applications, in particular in the field of biological applications. Recent tremendous developments in their synthesis open access to systems with complex architectures and compositions allowing for tailoring microgels with specific properties. At the same time state-of-the-art theoretical and simulation approaches offer deeper understanding of the behavior and structure of nano- and microgels under external influences and confinement at interfaces or at high volume fractions. Developments in the experimental analysis of nano- and microgels have become particularly important for structural investigations covering a broad range of length scales relevant to the internal structure, the overall size and shape, and interparticle interactions in concentrated samples. Here we provide an overview of the state-of-the-art, recent developments as well as emerging trends in the field of nano- and microgels. The following aspects build the focus of our discussion: tailoring (multi)functionality through synthesis; the role in biological and biomedical applications; the structure and properties as a model system, e.g., for densely packed arrangements in bulk and at interfaces; as well as the theory and computer simulation.

Nanogels and Microgels: From Model Colloids to Applications, Recent Developments, and Future Trends

3D computer simulations and experiments are employed to study random packings of compressible spherical grains under external confining stress. In the rigid ball limit, we find a continuous transition in which the stress vanishes as (straight phi-straight phi(c))(beta), where straight phi is the (solid phase) volume density. The value of straight phi(c) depends on whether the grains interact via only normal forces (giving rise to random close packings) or by a combination of normal and friction generated transverse forces (producing random loose packings). In both cases, near the transition, the system's response is controlled by localized force chains.

Packings of Compressible Granular Materials

Viral infection starts with a virus particle landing on a cell surface followed by penetration of the plasma membrane. Due to the difficulty of measuring the rapid motion of small-sized virus particles on the membrane, little is known about how a virus particle reaches an endocytic site after landing at a random location. Here, we use coherent brightfield (COBRI) microscopy to investigate early stage viral infection with ultrahigh spatiotemporal resolution. By detecting intrinsic scattered light via imaging-based interferometry, COBRI microscopy allows us to track the motion of a single vaccinia virus particle with nanometer spatial precision (<3 nm) in 3D and microsecond temporal resolution (up to 100,000 frames per second). We explore the possibility of differentiating the virus signal from cell background based on their distinct spatial and temporal behaviors via digital image processing. Through image postprocessing, relatively stationary background scattering of cellular structures is effectively rem...

Coherent Brightfield Microscopy Provides the Spatiotemporal Resolution To Study Early Stage Viral Infection in Live Cells.

An important goal of self-assembly is to achieve a preprogrammed structure with high fidelity. Here, we control the valence of DNA-functionalized emulsions to make linear and branched model polymers, or "colloidomers." The distribution of cluster sizes is consistent with a polymerization process in which the droplets achieve their prescribed valence. Conformational statistics reveal that the chains are freely jointed, so that the Kuhn length is close to one bead diameter. The end-to-end length scales with the number of bonds N as N^{ν}, where ν≈3/4, in agreement with the Flory theory in two dimensions. The chain diffusion coefficient D approximately scales as D∝N^{-ν}, as predicted by the Zimm model. Unlike molecular polymers, colloidomers can be repeatedly assembled and disassembled under temperature cycling, allowing for reconfigurable, responsive matter.

Freely Jointed Polymers Made of Droplets.

Accurate distance measurement in 3D confocal microscopy is important for quantitative analysis, volume visualization and image restoration. However, axial distances can be distorted by both the point spread function (PSF) and by a refractive-index mismatch between the sample and immersion liquid, which are difficult to separate. Additionally, accurate calibration of the axial distances in confocal microscopy remains cumbersome, although several high-end methods exist. In this paper we present two methods to calibrate axial distances in 3D confocal microscopy that are both accurate and easily implemented. With these methods, we measured axial scaling factors as a function of refractive-index mismatch for high-aperture confocal microscopy imaging. We found that our scaling factors are almost completely linearly dependent on refractive index and that they were in good agreement with theoretical predictions that take the full vectorial properties of light into account. There was however a strong deviation with the theoretical predictions using (high-angle) geometrical optics, which predict much lower scaling factors. As an illustration, we measured the PSF of a correctly calibrated point-scanning confocal microscope and showed that a nearly index-matched, micron-sized spherical object is still significantly elongated due to this PSF, which signifies that care has to be taken when determining axial calibration or axial scaling using such particles.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/jmi.12194

Methods to calibrate and scale axial distances in confocal microscopy as a function of refractive index

We introduce a new experimental method to encapsulate and release oils and fluorescent molecules into preformed elastic colloidal microcapsules of polydimethylsiloxane (PDMS)-filled siloxane shells, which are cross-linked with tetraethoxysilane. The method uses controlled buckling, where the volume of the capsule is reduced by dissolving the PDMS oil inside the capsule by surfactant micelles. This results in a change in the morphology of the capsule that depends on the ratio of shell thickness to total particle radius (d/Rt). Microcapsules of d/Rt in the range 0.007–0.05 formed microbowls upon decreasing the inner volume. The amount of oil released or dissolved by the micelles can be directly related to the concentration of surfactant. By tuning the amount of oil released, we can make microbowls of variable depth. In addition, we demonstrate that the microbowls can be further used to load different oils like silicone oil, hydrocarbons, and apolar dyes. The elasticity of the capsule wall and the leftover P...

/pdf/unloading-and-reloading-colloidal-microcapsules-with-apolar-1o364epn8j.pdf

Unloading and reloading colloidal microcapsules with apolar solutions by controlled and reversible buckling.

We report several facile, surfactant-free methods to prepare monodisperse polydimethylsiloxane (PDMS) droplets in the size range 3–8 μm in water. These methods, of which the pros and cons are discussed, are extensions of a procedure described before by our group which focused on smaller droplet sizes. The PDMS oil droplets are formed by ammonia catalyzed hydrolysis and condensation of the monomer dimethyldiethoxysilane (DMDES) in water. One of the methods entails a seeded growth procedure in which other oils, such as lower molecular weight hydrocarbons, were found to be able to swell the PDMS droplets if their solubility in water was higher than that of the seed droplets. This way, larger droplets with mixed composition could be prepared. It also turned out to be possible to load the monodisperse droplets with an oil soluble dye. The droplets could be coated with an elastic, partially permeable, shell formed by cross-linking the PDMS with tetraethoxysilane (TES) in the presence of poly(vinylpyrrolidone) (...

https://pubs.acs.org/doi/pdf/10.1021/cm504504t

Bulk Scale Synthesis of Monodisperse PDMS Droplets above 3 μm and Their Encapsulation by Elastic Shells

Cellulose microfibrils (CMFs) are an important nanoscale building block in many novel biobased functional materials. The spatial nano- and microscale organization of the CMFs is a crucial factor for defining the properties of these materials. Here, we report for the first time a direct three-dimensional (3D) real-space analysis of individual CMFs and their networks formed after ultrahigh-shear-induced transient deagglomeration and self-assembly in a solvent. Using point-scanning confocal microscopy combined with tracking the centerlines of the fibrils and their junctions by a stretching open active contours method, we reveal that dispersions of the native CMFs assemble into highly heterogeneous networks of individual fibrils and bundles. The average network mesh size decreases with increasing CMF volume fraction. The cross-sectional width and the average length between the twists in the ribbon-shaped CMFs are directly determined and compared well with that of fibrils in the dried state. Finally, the gener...

Revealing and Quantifying the Three-Dimensional Nano- and Microscale Structures in Self-Assembled Cellulose Microfibrils in Dispersions

Accurate distance measurement in 3D confocal microscopy is important for quantitative analysis, volume visualization and image restoration. However, axial distances can be distorted by both the point spread function and by a refractive-index mismatch between the sample and immersion liquid, which are difficult to separate. Additionally, accurate calibration of the axial distances in confocal microscopy remains cumbersome, although several high-end methods exist. In this paper we present two methods to calibrate axial distances in 3D confocal microscopy that are both accurate and easily implemented. With these methods, we measured axial scaling factors as a function of refractive-index mismatch for high-aperture confocal microscopy imaging. We found that our scaling factors are almost completely linearly dependent on refractive index and that they were in good agreement with theoretical predictions that take the full vectorial properties of light into account. There was however a strong deviation with the theoretical predictions using (high-angle) geometrical optics, which predict much lower scaling factors. As an illustration, we measured the point-spread-function of a point-scanning confocal microscope and showed that an index-matched, micron-sized spherical object is still significantly elongated due to this PSF, which confirms that single micron-sized spheres are not well suited to determine accurate axial calibration nor axial scaling.

Jissy Jose

Papers

Methods to calibrate and scale axial distances in confocal microscopy as a function of refractive index

Unloading and reloading colloidal microcapsules with apolar solutions by controlled and reversible buckling.

Bulk Scale Synthesis of Monodisperse PDMS Droplets above 3 μm and Their Encapsulation by Elastic Shells

Revealing and Quantifying the Three-Dimensional Nano- and Microscale Structures in Self-Assembled Cellulose Microfibrils in Dispersions

Methods to calibrate and scale axial distances in confocal microscopy as a function of refractive index