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"Light scattering"
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Structural Characterization of Biomaterials by Means of Small Angle X-rays and Neutron Scattering (SAXS and SANS), and Light Scattering Experiments
Scattering techniques represent non-invasive experimental approaches and powerful tools for the investigation of structure and conformation of biomaterial systems in a wide range of distances, ranging from the nanometric to micrometric scale. More specifically, small-angle X-rays and neutron scattering and light scattering techniques represent well-established experimental techniques for the investigation of the structural properties of biomaterials and, through the use of suitable models, they allow to study and mimic various biological systems under physiologically relevant conditions. They provide the ensemble averaged (and then statistically relevant) information under in situ and operando conditions, and represent useful tools complementary to the various traditional imaging techniques that, on the contrary, reveal more local structural information. Together with the classical structure characterization approaches, we introduce the basic concepts that make it possible to examine inter-particles interactions, and to study the growth processes and conformational changes in nanostructures, which have become increasingly relevant for an accurate understanding and prediction of various mechanisms in the fields of biotechnology and nanotechnology. The upgrade of the various scattering techniques, such as the contrast variation or time resolved experiments, offers unique opportunities to study the nano- and mesoscopic structure and their evolution with time in a way not accessible by other techniques. For this reason, highly performant instruments are installed at most of the facility research centers worldwide. These new insights allow to largely ameliorate the control of (chemico-physical and biologic) processes of complex (bio-)materials at the molecular length scales, and open a full potential for the development and engineering of a variety of nano-scale biomaterials for advanced applications.
Journal Article
The airborne lifetime of small speech droplets and their potential importance in SARS-CoV-2 transmission
Speech droplets generated by asymptomatic carriers of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are increasingly considered to be a likely mode of disease transmission. Highly sensitive laser light scattering observations have revealed that loud speech can emit thousands of oral fluid droplets per second. In a closed, stagnant air environment, they disappear from the window of view with time constants in the range of 8 to 14 min, which corresponds to droplet nuclei of ca. 4 μm diameter, or 12- to 21-μm droplets prior to dehydration. These observations confirm that there is a substantial probability that normal speaking causes airborne virus transmission in confined environments.
Journal Article
Nanoparticle number concentration measurements by multi-angle dynamic light scattering
We present the use of Multi-angle Dynamic Light Scattering (MADLS®) for the measurement of nanoparticle concentration. We describe the theory of the method and its application to nanoparticles made of gold, silica and polystyrene, with diameters ranging from 30 to 400 nm, and demonstrate some of the limitations with particles of sizes 500 nm and above. We evaluate the method accuracy, linearity and reproducibility, as well as the operational nanoparticle concentration and size range. We show that the concentration working range depends on the material’s optical properties, size and concentration. Here it is shown that the level of accuracy that can be expected for the concentration of particles is typically within 50% of the nominal value across a range of materials and sizes and, for some samples, within 20%. The repeatability of the measurements, in terms of relative standard deviation, is typically below 30%. A linearity of within 40% across a concentration range of 3·108 to 2·1011 mL−1 for concentration measurements was also demonstrated by using gold nanoparticles and gravimetric dilutions for method validation. Overall, we show that MADLS® is a rapid and straightforward method for the reproducible measurement of particle concentration, as well as size, requiring minimal sample preparation, without the need to calibrate using a pre-determined concentration series, and applicable to a broad range of materials. These features make it an ideal tool to support both development and quality control of particle materials for a broad range of applications.
Journal Article
A review of exosome separation techniques and characterization of B16-F10 mouse melanoma exosomes with AF4-UV-MALS-DLS-TEM
Exosomes participate in cancer metastasis, but studying them presents unique challenges as a result of their small size and purification difficulties. Asymmetrical field flow fractionation with in-line ultraviolet absorbance, dynamic light scattering, and multi-angle light scattering was applied to the size separation and characterization of non-labeled B16-F10 exosomes from an aggressive mouse melanoma cell culture line. Fractions were collected and further analyzed using batch mode dynamic light scattering, transmission electron microscopy and compared with known size standards. Fractogram peak positions and computed radii show good agreement between samples and across fractions. Ultraviolet absorbance fractograms in combination with transmission electron micrographs were able to resolve subtle heterogeneity of vesicle retention times between separate batches of B16-F10 exosomes collected several weeks apart. Further, asymmetrical field flow fractionation also effectively separated B16-F10 exosomes into vesicle subpopulations by size. Overall, the flow field flow fractionation instrument combined with multiple detectors was able to rapidly characterize and separate exosomes to a degree not previously demonstrated. These approaches have the potential to facilitate a greater understanding of exosome function by subtype, as well as ultimately allow for “label-free” isolation of large scale clinical exosomes for the purpose of developing future exosome-based diagnostics and therapeutics.
Figure
Flow path of exosome sample through the asymmetrical field flow fractionation instrument, detectors, and transmission electron microscope.
Journal Article
Scattered Light Imaging: Resolving the substructure of nerve fiber crossings in whole brain sections with micrometer resolution
•Scattered Light Imaging (SLI) reveals individual nerve fiber directions in the brain.•The simple prototypic setup contains only a standard LED light source and a camera.•SLI reconstructs multiple crossing nerve fiber directions within each image pixel.•We measured various brain sections (rodent/monkey/human) with micrometer resolution.•We validated our results against simulated/measured scattering patterns and 3D-PLI.
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For developing a detailed network model of the brain based on image reconstructions, it is necessary to spatially resolve crossing nerve fibers. The accuracy hereby depends on many factors, including the spatial resolution of the imaging technique. 3D Polarized Light Imaging (3D-PLI) allows the three-dimensional reconstruction of nerve fiber tracts in whole brain sections with micrometer in-plane resolution, but leaves uncertainties in pixels containing crossing fibers. Here we introduce Scattered Light Imaging (SLI) to resolve the substructure of nerve fiber crossings. The measurement is performed on the same unstained histological brain sections as in 3D-PLI. By illuminating the brain sections from different angles and measuring the transmitted (scattered) light under normal incidence, light intensity profiles are obtained that are characteristic for the underlying brain tissue structure. We have developed a fully automated evaluation of the intensity profiles, allowing the user to extract various characteristics, like the individual directions of in-plane crossing nerve fibers, for each image pixel at once. We validate the reconstructed nerve fiber directions against results from previous simulation studies, scatterometry measurements, and fiber directions obtained from 3D-PLI. We demonstrate in different brain samples (human optic tracts, vervet monkey brain, rat brain) that the 2D fiber directions can be reliably reconstructed for up to three crossing nerve fiber bundles in each image pixel with an in-plane resolution of up to 6.5 μm. We show that SLI also yields reliable fiber directions in brain regions with low 3D-PLI signals coming from regions with a low density of myelinated nerve fibers or out-of-plane fibers. This makes Scattered Light Imaging a promising new imaging technique, providing crucial information about the organization of crossing nerve fibers in the brain.
Journal Article
Light scattering by particles in water : theoretical and experimental foundations
Light scattering-based methods are used to characterize small particles suspended in water in a wide range of disciplines ranging from oceanography, through medicine, to industry. The scope and accuracy of these methods steadily increases with the progress in light scattering research. This book focuses on the theoretical and experimental foundations of the study and modeling of light scattering by particles in water and critically evaluates the key constraints of light scattering models. It begins with a brief review of the relevant theoretical fundamentals of the interaction of light with condensed matter, followed by an extended discussion of the basic optical properties of pure water and seawater and the physical principles that explain them. The book continues with a discussion of key optical features of the pure water/seawater and the most common components of natural waters. In order to clarify and put in focus some of the basic physical principles and most important features of the experimental data on light scattering by particles in water, the authors employ simple models. The book concludes with extensive critical reviews of the experimental constraints of light scattering models: results of measurements of light scattering and of the key properties of the particles: size distribution, refractive index (composition), structure, and shape. These reviews guide the reader through literature scattered among more than 210 scientific journals and periodicals which represent a wide range of disciplines. A special emphasis is put on the methods of measuring both light scattering and the relevant properties of the particles, because principles of these methods may affect interpretation and applicability of the results. The book includes extensive guides to literature on light scattering data and instrumentation design, as well as on the data for size distributions, refractive indices, and shapes typical of particles in natural waters. It also features a comprehensive index, numerous cross-references, and a reference list with over 1370 entries. An errata sheet for this work can be found at: http://www.tpdsci.com/Ref/Jonasz_M_2007_LightScatE.php *Extensive reference section provides handy compilations of knowledge on the designs of light scattering meters, sources of experimental data, and more *Worked exercises and examples throughout
eBook
Optical Coatings and Thermal Noise in Precision Measurement
Thermal noise from optical coatings is a growing area of concern and overcoming limits to the sensitivity of high precision measurements by thermal noise is one of the greatest challenges faced by experimental physicists. In this timely book, internationally renowned scientists and engineers examine our current theoretical and experimental understanding. Beginning with the theory of thermal noise in mirrors and substrates, subsequent chapters discuss the technology of depositing coatings and state-of-the-art dielectric coating techniques used in precision measurement. Applications and remedies for noise reduction are also covered. Individual chapters are dedicated to specific fields where coating thermal noise is a particular concern, including the areas of quantum optics/optomechanics, gravitational wave detection, precision timing, high-precision laser stabilisation via optical cavities and cavity quantum electrodynamics. While providing full mathematical detail, the text avoids field-specific jargon, making it a valuable resource for readers with varied backgrounds in modern optics.
eBook
Dynamic light scattering distributions by any means
Dynamic light scattering (DLS) is an essential technique for nanoparticle size analysis and has been employed extensively for decades, but despite its long history and popularity, the choice of weighting and mean of the size distribution often appears to be picked ad hoc to bring the results into agreement with other methods and expectations by any means necessary. Here, we critically discuss the application of DLS for nanoparticle characterization and provide much-needed clarification for ambiguities in the mean-value practice of commercial DLS software and documentary standards. We address the misleading way DLS size distributions are often presented, that is, as a logarithmically scaled histogram of measured relative quantities. Central values obtained incautiously from this representation often lead to significant interpretation errors. Through the measurement of monomodal nanoparticle samples having an extensive range of sizes (5 to 250 nm) and polydispersity, we similarly demonstrate that the default outputs of a frequently used DLS inversion method are ill chosen, as they are regularizer-dependent and significantly deviate from the cumulant z-average size. The resulting discrepancies are typically larger than 15% depending on the polydispersity index of the samples. We explicitly identify and validate the harmonic mean as the central value of the intensity-weighted DLS size distribution that expresses the inversion results consistently with the cumulant results. We also investigate the extent to which the DLS polydispersity descriptors are representative of the distributional quality and find them to be unreliable and misleading, both for monodisperse reference materials and broad-distribution biomedical nanoparticles. These results overall are intended to bring essential improvements and to stimulate reexamination of the metrological capabilities and role of DLS in nanoparticle characterization.
Journal Article
Optical focusing deep inside dynamic scattering media with near-infrared time-reversed ultrasonically encoded (TRUE) light
Focusing light deep inside living tissue has not been achieved despite its promise to play a central role in biomedical imaging, optical manipulation and therapy. To address this challenge, internal-guide-star-based wavefront engineering techniques—for example, time-reversed ultrasonically encoded (TRUE) optical focusing—were developed. The speeds of these techniques, however, were limited to no greater than 1 Hz, preventing them from
in vivo
applications. Here we improve the speed of optical focusing deep inside scattering media by two orders of magnitude, and focus diffuse light inside a dynamic scattering medium having a speckle correlation time as short as 5.6 ms, typical of living tissue. By imaging a target, we demonstrate the first focusing of diffuse light inside a dynamic scattering medium containing living tissue. Since the achieved focusing speed approaches the tissue decorrelation rate, this work is an important step towards
in vivo
deep tissue noninvasive optical imaging, optogenetics and photodynamic therapy.
Shaping the incident wavefront allows optical focusing deep inside scattering media, but its application in dynamic media is hindered by its low speed. Here, Liu
et al
. improve the speed by two orders of magnitude and demonstrate
in vivo
light focusing inside dynamic scattering media.
Journal Article