The interaction between wheat germ agglutinin and membrane incorporated glycophorin A. An optical binding study

A novel integrated optical technique is used to monitor the kinetics of incorporation of glycophorin A (GPA) from solution into a planar dimyristoylphosphatidylcholine-cholesterol bilayer membrane, and the subsequent binding of wheat germ agglutinin (WGA) to the membrane-incorporated GPA. The technique significantly improves the attainable accuracy of kinetic measurements. The number of bound molecules can be determined to a precision of ca ± 80 mol µm^–2. Our results show that GPA incorporates spontaneously into the bilayer. Binding of WGA to GPA is optimal in the presence of human serum albumin, and can be reversed byN-acetyl-d-glucosamine. The kinetics of the binding are consistent with the presence of two classes of kinetically distinguishable binding sites with association rates of 2.0×10⁴ and 9.6×10² M^–1 s^–1, and dissociation rates of 2.7×10^–3 s^–1 and <10^–5 s^–1, respectively. A stoichiometry of 4 WGA monomers per GPA monomer was determined as characteristic of the overall binding interaction.Abbreviations DMPC dimyristoylphosphatidylcholine - GlcNAc N-acetyl-d-glucosamine - GPA glycophorin A - HSA human serum albumin - NeuNAc N-acetyl-d-neuraminic acid - TE transverse electric - TM transverse magnetic - WGA wheat germ agglutinin     

Organometallic complexes for nonlinear optics. Part 36. Quadratic and cubic optical nonlinearities of 4-fluorophenylethynyl- and 4-nitro-(E)-stilbenylethynylruthenium complexes

The complexes trans-[Ru(CC-4-C₆H₄F)X(dppe)₂] [X = Cl (1), CCPh (2), CC-4-C₆H₄NO₂ (3)], trans-[Ru{CC-4-C₆H₄-(E)-CHCH-4-C₆H₄NO₂}X(dppe)₂] [X = CCPh (4), CC-4-C₆H₄CCPh (5)], and [C₆H₃-1,3-{CC-trans-[RuCl(dppe)₂]}₂-5-(CC-4-C₆H₄F)] (6) have been synthesized and the identity of 1 confirmed by a single-crystal X-ray diffraction study. Cyclic voltammetry reveals a metal-centered oxidation, the potential of which is largely invariant on alkynyl ligand replacement across the series 1-5; the diruthenium complex 6 shows two oxidation processes, consistent with weakly interacting metal centers. Hyper-Rayleigh scattering (HRS) studies at 1064 nm using ns pulses suggest quadratic nonlinearities for 3-5 that are amongst the largest thus far for organometallic complexes, a trend maintained with the two-level-corrected data. HRS studies at 800 nm using fs pulses and amplitude modulation to remove multi-photon fluorescence contributions reveal significant fluorescence-free nonlinearities for 3-5; the frequency-independent nonlinearities calculated from the 800 nm results are suggestive of fluorescence contributions to the 1064 nm data. Z-scan studies at 820 nm reveal cubic nonlinearities that increase with the size of the π-system, although error margins are significant.     

A novel form of Total Internal Reflection Fluorescence Microscopy (LG-TIRFM) reveals different and independent lipid raft domains in living cells

In the present study we have applied a novel form of Total Internal Reflection Fluorescence Microscopy (LG-TIRFM) in combination with fluorescently labeled cholera toxin to the study of lipid rafts dynamics in living cells. We demonstrate the usefulness of such approach by showing the dynamic formation/disaggregation of islands of cholera toxin on the surface of cells. Using multicolor LG-TIRFM with co-localization studies we show for the first time that two receptors previously identified as constituents of lipid rafts are found on different and independent “raft domains” on the cell plasma membrane. Furthermore, LG-TIRFM studies revealed limited association and dissociation of both domains overtime on different areas of the plasma membrane. The implications of different “raft domains” on cell physiology are discussed.     

Biomedical Image Processing with Containers and Deep Learning: An Automated Analysis Pipeline

Here, a streamlined, scalable, laboratory approach is discussed that enables medium‐to‐large dataset analysis. The presented approach combines data management, artificial intelligence, containerization, cluster orchestration, and quality control in a unified analytic pipeline. The unique combination of these individual building blocks creates a new and powerful analysis approach that can readily be applied to medium‐to‐large datasets by researchers to accelerate the pace of research. The proposed framework is applied to a project that counts the number of plasmonic nanoparticles bound to peripheral blood mononuclear cells in dark‐field microscopy images. By using the techniques presented in this article, the images are automatically processed overnight, without user interaction, streamlining the path from experiment to conclusions.     

用侧向光源进行前列腺光声扫描成像的研究

光声成像技术是近年来兴起的前列腺早期检测与成像的新技术.前列腺组织的结构特征需要一种无创的且光穿透深度足够大的光声激发方式.本文设计了一种可用于经由尿道对前列腺光辐照的侧向光源,并结合直肠处长焦区聚焦式超声换能器的光声探测展开了前列腺仿体的光声扫描成像.结果表明,侧向光源有利于单侧吸收体的定位和成像,具有较好的成像范围和深度,结合侧向光源的旋转可实现全方位成像.因此该侧向光源有望成为早期前列腺肿瘤光声成像技术中一种新型的光源结构,具有重要意义.     

Zero-cost modification of bright field microscopes for imaging phase gradient on cells: Schlieren optics

A simple, zero-cost, reversible modification of a bright field microscope permits visualization of phase gradients in cells by transmitted illumination, yielding a Nomarski-like effect. This modification, based on schlieren optics, is simultaneously compatible with high-aperture epi-illumination fluorescence excitation. For many objectives that are intended for use in fluorescence work, but are unavailable in phase contrast versions, the modification provides a simple means for locating cells in culture with good image contrast and resolution.     

ESTIMATION OF CHLOROPHYLL a FROM TIME SERIES MEASUREMENTS OF HIGH SPECTRAL RESOLUTION REFLECTANCE IN AN EUTROPHIC LAKE

We acquired high spectral resolution reflectance data in Carter Lake, a eutrophic oxbow on the Iowa–Nebraska border, from April 1995 to April 1996. Chlorophyll a, total seston, sestonic organic matter, Secchi depth, and nephelometric turbidity were determined for each respective spectral measurement. Changes in algal taxonomic structure and abundance coincided with the development and senescence of a midsummer through autumn bloom of Anabaena. Taxonomic structure was more diverse in late winter and spring when Synedra sp. (diatom) and several chlorophytes and dinoflagellates were present. Overall, chlorophyll a varied from about 20 to 280 μg·L^?1, Secchi transparency from 18 to 74 cm, and seston dry weight from 11 to 48 mg·L^?1 in February and September, respectively. Particulate matter completely dominated lake water light attenuation. Dissolved organic matter had low optical activity. The most sensitive spectral feature to variation in chlorophyll a concentration was the magnitude of the scattering peak near 700 nm. The 700-nm peak correlated to chlorophyll concentration through the relationships between algal pigment absorption near 670 nm and the cell biomass and surface-related scattering signal in the near infrared. An algorithm relating the height of the 700-nm reflectance peak above a reference baseline between 670 and 850 nm to chlorophyll a was accurate and robust despite large variations in optical constituents caused by both strong seasonality in the algal system and short-term variations in seston from wind-induced sediment resuspension. The present algorithms were successfully used in other systems with different seasonality and productivity patterns. The coefficients of the models relating chlorophyll a and spectral reflectance variables appeared to be ecosystem specific: both the intercept and slope for the models in this study were moderately lower than for several other recently published results. We validated our algorithm coefficients with a second, independent dataset. The standard error for chlorophyll a prediction was ±28 μg·L^?1.     

Eye morphology and optics of the double-eyed mysid Euchaetomera typica

The structure and optics of the mesopelagic double-eyed mysid crustacean Euchaetomera typica Sars, 1884 are described for the first time. The lateral eye is a typical refracting superposition eye with a wide field of view (172°) and low resolution (interommatidial angle of 7.3°). The antero-dorsal part of the eye is elongated due to the extension of the clear zone. This dorsal eye has a restricted field of view (33°) but much higher resolution (1.5°). The dorsal eye also uses refracting superposition optics, although the optical array is unusual as many of the peripheral ommatidia lack crystalline cones. The centre of curvature of the cornea is in front of the flattened rhabdom layer whereas the axes of the crystalline cones are centred on a point about twice as deep as the rhabdom layer. This results in a well-focused eye, free of spherical aberration. There is a remarkable similarity in eye structure between this species and some mesopelagic double-eyed euphausiid crustaceans.     

Inside Cover: Fiber‐based fluorescence lifetime imaging of recellularization processes on vascular tissue constructs (J. Biophotonics 9/2018)

Tissue autofluorescence provides fluorescence lifetime contrast between acellular tissue and that containing newly seeded cells. Fiber‐based fluorescence lifetime imaging (FLIm) can be used for tracking recellularization of engineered vascular grafts and potential matrix remodeling at large scale, without compromising sample integrity. FLIm cellular contrast was verified in a subset of samples seeded with eGFP‐labelled cells. Results suggests fiberbased FLIm is a suitable tool for monitoring recellularization of engineered tissue nondestructively. Further details can be found in the article by Alba Alfonso‐Garcia, Jeny Shklover, Benjamin E. Sherlock, et al. ( e201700391 ).