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Three‐dimensional quantitative phase imaging is an emerging method, which provides the 3D distribution of the refractive index (RI) and the dry mass in live and fixed cells as well as in tissues. However, an insufficiently answered question is the influence of chemical cell fixation procedures on the results of RI reconstructions. Therefore, this work is devoted to systematic investigations on the RI in cellular organelles of live and fixed cells including nucleus, nucleolus, nucleoplasm, and cytoplasm. The research was carried out on four different cell lines using a common paraformaldehyde (PFA)‐based fixation protocol. The selected cell types represent the diversity of mammalian cells and therefore the results presented provide a picture of fixation caused RI changes in a broader context. A commercial Tomocube HT‐1S device was used for 3D RI acquisition. The changes in the RI values after the fixation process are detected in the reconstructed phase distributions and amount to the order of 10−3. The RI values decrease and the observed RI changes are found to be different between various cell lines; however, all of them show the most significant loss in the nucleolus. In conclusion, our study demonstrates the evident need for standardized preparation procedures in phase tomographic measurements. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals LLC. on behalf of International Society for Advancement of Cytometry.

Besides water, raw methanol produced from steel mill gases within the Carbon2Chem® project contains dissolved carbon dioxide. Refractometry and densimetry were investigated as analysis methods to analyze raw methanol samples quickly and reliably. To verify existing calibration curves, a set of standard solutions from pure chemicals was produced. Experimental results differed significantly from published refractive index data, especially in the range of high methanol weight fractions, which are of particular interest for the Carbon2Chem® project.

Background Brix refractometry can be used to assess colostral immunoglobulin G (IgG) concentration, but studies identifying Brix percentages to detect high‐ and low‐IgG colostrum are lacking for beef cows and interlaboratory agreement is unknown. Objectives Evaluate Brix refractometer performance and interlaboratory agreement for assessing beef cow colostrum IgG concentration, including determination of thresholds to identify colostrum containing IgG concentrations

Digital holographic microscopy (DHM) is an important technique that may be used for quantitative phase imaging of unstained biological cell samples. Since the DHM technology is not commonly used in clinics or bioscience research labs, at present there is no well-accepted focusing criterion for unstained samples that users can follow while recording image plane digital holograms of cells. The usual sharpness metrics that are useful for auto-focusing of stained cells do not work well for unstained cells as there is no amplitude contrast. In this work, we report a practical method for estimating the best focus plane for unstained cells in the digital hologram domain. The method is based on an interesting observation that for the best focus plane the fringe pattern associated with individual unstained cells predominantly shows phase modulation effect in the form of bending of fringes and minimal amplitude modulation. This criterion when applied to unstained red blood cells shows that the central dip in the doughnut-like phase profile of cells is maximal in this plane. The proposed methodology is helpful for standardizing the usage of DHM technology across different users and application development efforts. LAY DESCRIPTION: Digital holographic microscopy (DHM) is slowly but steadily becoming an important microscopy modality and gaining acceptability for basic bio-science research as well as clinical usage. One of the important features of DHM is that it allows users to perform quantitative imaging of unstained transparent cells. Instead of using dyes or fluorescent labelling, DHM systems use quantitative phase as a contrast mechanism which depends on the natural refractive index variation within the cell samples. Since minimal wet lab processing is required in order to image cell samples with a DHM, cells can be imaged in their natural state. While DHM is gaining popularity among users, the imaging protocols across the labs or users need to be standardized in order to make sure that the same quantitative phase parameters are used for tasks such as quantitative phased based cell classification. One of the important operational tasks for any microscopy work is to focus the sample under study. While focusing comes naturally to users of brightfield microscopes based on image contrast, the focusing is not straightforward when samples are unstained so that they do not offer any amplitude contrast. When performing quantitative phase imaging, defocus can actually change the phase profile of the cell due to near-zone (Fresnel) diffraction effects. So unless a standardized focusing methodology is used, it will be difficult for multiple DHM users (potentially at different sites) to agree on quantitative results out of their phase images. DHM literature has prior works which perform numerical focusing of recovered complex wave-field in the hologram plane to find the best focus plane. However such methods are not user friendly and do not allow user the same focusing experience as in a brightfield microscope. The numerical focusing is therefore a reasonably good method for an optics researcher but not necessarily so for a microscopy technician looking at cell samples with a DHM system in a clinical setting. The present work provides a simple focusing criterion for unstained samples that works directly in the hologram domain. The technique is based on an interesting observation that the when an unstained cell sample is in the best-focus plane, its corresponding hologram (or fringe pattern) predominantly shows phase modulation manifested by bending of fringes at the location of the cell. This criterion can be converted into a simple numerical method as we have used to find the best-focus plane using a stack of through focus holograms. We believe that the technique can be used manually by visually observing the holograms or can be converted to an auto-focus algorithm for a motorized DHM system.

In recent years, production of biosynthesized human milk oligosaccharides (HMOs) has become scalable to industrial standards. As a result, infant formula fortified with 2’‐fucosyllactose (2’‐FL), the most abundant HMO in human breast milk, is now commercially available. 2’‐FL and 3‐fucosyllactose (3‐FL), another abundant HMO, are thought to be beneficial for infant health and development. Products containing HMOs are projected to expand in the future, showing the need for robust, easily applicable analytical methods for the quantitative assessment of HMOs in different food applications. We present here a validated high‐performance liquid chromatography method for the quantification of 2’‐FL and 3‐FL in whole milk, infant formula, and cereal bars. The sample preparation was simple dispersion and extraction of the sample. The samples were analyzed by hydrophilic interaction liquid chromatography with refractive index detection and a runtime of 19 min. The method had a high degree of linearity (R 2 > 0.9995) in the range 0.2 to 12 mg/mL. The recovery for 2’‐FL was 88% to 105% and for 3‐FL 94% to 112%. The limit of detection (LOD) for whole milk was 0.1 mg/mL for 2’‐FL and 0.2 mg/mL for 3‐FL. In infant formula and cereal bars, the LOD was 0.6 mg/g for both 2’‐FL and 3‐FL. To show the practical application of this method, it was successfully utilized in stability studies of 2’‐FL and 3‐FL in whole milk, UHT milk, and yoghurt. The method provides a means of simultaneous and robust quantification of 2’‐FL and 3‐FL in various food matrices with high accuracy and high reproducibility. Practical Application 2’‐Fucosyllactose (2’‐FL) and 3‐fucosyllactose (3‐FL) are two of the most abundant human milk oligosaccharides (HMOs) present in human breast milk. We present a fast HPLC method for the robust quantification of these two compounds in infant formula, whole milk, UHT milk, cereal bars, and yoghurt. This method can easily be set up by food producers and researchers to analyze the dosage of 2’‐FL and 3‐FL in their product or perform shelf life studies in different food applications.

Fourier ptychographic diffraction tomography (FPDT) is a recently developed label-free computational microscopy technique that retrieves high-resolution and large-field 3D tomograms by synthesizing a set of low-resolution intensity images obtained with a low numerical aperture (NA) objective. However, in order to ensure sufficient overlap of Ewald spheres in 3D Fourier space, conventional FPDT requires thousands of intensity measurements and consumes a significant amount of time for stable convergence of the iterative algorithm. Herein, we present accelerated Fourier ptychographic diffraction tomography (aFPDT), which combines sparse annular LED illuminations and multiplexing illumination to significantly decrease data amount and achieve computational acceleration of 3D RI tomography. Compared with existing FPDT technique, the equivalent high-resolution 3D RI results are obtained using aFPDT with reducing data requirement by more than 40 times. The validity of the proposed method is experimentally demonstrated on control samples and various biological cells, including polystyrene beads, unicellular algae, and clustered HeLa cells in a large field of view. With the capability for high-resolution and high-throughput 3D imaging using small amounts of data, aFPDT has the potential to further advance its widespread applications in biomedicine. This article is protected by copyright. All rights reserved.

Sugar concentration in floral nectars is an assessment required in several diverse fields of application. The widely used analysis, consisting of nectar extraction with a microcapillary and sugar concentration measurement with a light refractometer, is not reliable when the nectar is secreted in small quantities, unextractable with a microcapillary. Ancillary methods adopted in such cases are destructive, rather complicated and often provide much less precise and accurate results. The microscopic-size, low cost and biocompatibility of optical fibers were exploited to deliver light directly inside the flower with minimal invasiveness and measure instantaneously the refractometric properties of the nectar without extracting it. After comparing the new and old methods using two known nectariferous species, the new approach was validated on Primula palinuri, whose nectar is unextractable with microcapillaries. The fiber-optic probe was able to measure the nectar refractive index in P. palinuri flowers making it possible to highlight a previously undetected significant trend of the sugar concentration throughout the long anthesis of the single flowers. Changes in nectar concentrations are similar in both longistylous and brevistylous flowers. The fiber-optic refractometer is an advancement of light refractometer analysis. Further customization of the laboratory set-up into portable equipment will boost applications.

Background Urine specific gravity (USG) is an integral part of the urinalysis and a key component of many clinical decisions, and fluctuations in USG have the potential to impact case management. Objectives To determine the intraindividual variability of first morning USG results in healthy dogs. Animals One hundred three healthy client-owned dogs. Methods Dogs were deemed healthy based on clinical history and physical examination findings. Repeated USG measurements were performed over the course of 2 weeks. Three urine samples were collected each week for a total of 6 samples per dog. Sample collection was distributed evenly throughout the week. Urine samples were acquired immediately upon waking and before any ingestion of liquids, food, or exertion of physical activity in the dogs. All measurements were made using the same Misco digital refractometer. Results Intraindividual USG was variable over the course of the study. The mean difference between the minimum and maximum USG for each dog was 0.015 (SD, 0.007). The within-week difference between the minimum and maximum USG was less than over the complete 2-week study (0.009 [SD 0.006] for week 1 and 0.010 [SD 0.007] for week 2). The mean coefficient of variance across all 6 time points was 15.4% (SD 8.97%). Conclusions and clinical importance Clinically important variation occurs in USG in healthy animals and might impact clinical decision-making when diagnostic cutoff points are utilized. Clinicians should be aware of inherent variability in this clinical variable when analyzing results.

We developed and evaluated the properties of in-house urine reference materials for the verification of laboratory refractometers, which are frequently used in clinical chemistry and doping testing laboratories. Urine was gathered from 26 healthy volunteers (16 male 30 ± 5 years old and 10 female 29 ± 4 years old), from which two urine batches were obtained: one with a low specific gravity (1.012± 0.003) and the other with a high specific gravity (1.027 ± 0.003). Homogeneity studies were conducted over 20 consecutive days. For short-term stability studies, aliquots of both urine batches were stored at -20 ± 2°C; 3 ± 2°C; 20 ± 2°C; 45 ± 2°C for 0, 2, 7, 14 and 35 days, under both light and dark conditions. Similarly, another study was conducted to measure the long-term stability of urine at -20 ± 2°C, over a 24-month evaluation period. Our data showed that the urine was homogeneous and stable at -20 ± 2°C, 3 ± 2°C, 20 ± 2°C, and 45 ± 2°C under both light and dark conditions. In all cases, the urine was evaluated by specific gravity and no statistically significant differences (p ≤ 0.05) were recorded. Additionally, a proficiency test was conducted in collaboration with 15 ISO/IEC 17025 accredited laboratories, and z-scores and performance factors were evaluated. These data indicate that this material could be used for the verification of refractometers.

The ability to quantitatively monitor various cellular activities is critical for understanding their biological functions and the therapeutic response of cells to drugs. Unfortunately, existing approaches such as fluorescent staining and impedance-based methods are often hindered by their multiple time-consuming preparation steps, sophisticated labeling procedures, and complicated apparatus. The cost-effective, monolithic gallium nitride (GaN) photonic chip has been demonstrated as an ultrasensitive and ultracompact optical refractometer in a previous work, but it has never been applied to cell studies. Here, for the first time, the so-called GaN chipscope is proposed to quantitatively monitor the progression of different intracellular processes in a label-free manner. Specifically, the GaN-based monolithic chip enables not only a photoelectric readout of cellular/subcellular refractive index changes but also the direct imaging of cellular/subcellular ultrastructural features using a customized differential interference contrast (DIC) microscope. The miniaturized chipscope adopts an ultracompact design, which can be readily mounted with conventional cell culture dishes and placed inside standard cell incubators for real-time observation of cell activities. As a proof-of-concept demonstration, its applications are explored in 1) cell adhesion dynamics monitoring, 2) drug screening, and 3) cell differentiation studies, highlighting its potential in broad fundamental cell biology studies as well as in clinical applications.

The high refractive index aromatic compound, binaphthol (BINOL), is readily incorporated into silicone polymer chains using the Piers-Rubinsztajn (PR) reaction; alternating and random linear copolymers, and elastomers are available. The highest refractive index (RI) materials are BINOL rich. It is not possible to directly make high refractive index linear polymers with very short HSi-capped, telechelic silicone chains, as they do not react cleanly. However, chain extending short vinyl-capped BINOL macromers with simple arylsilanes using hydrosilylation leads to polymers with a molar mass of up to 8000 and refractive indices of up to 1.58. Elastomers are prepared using similar processes. The reactions are facile to practice and suggest BINOL can be harnessed in these and other processes to augment RI.

Background Refractometric determination of total protein (TP) in synovial fluid (SF) is commonly used for diagnosis and monitoring of synovial sepsis in horses. Previous studies have shown that elevated concentrations of certain anticoagulants may overestimate refractometric determination of TP concentration. Objectives The aim of the study was to evaluate the effect of different concentrations of dipotassium EDTA (K2EDTA) and lithium heparin (LH) on TP determination by using a hand-held refractometer in equine synovial fluid. Study design Cross-section observational study. Methods Thirty samples of synovial fluid obtained from 22 horses with different synovial conditions were collected. Synovial fluid samples were separated into different aliquots and placed in commercially available collection tubes containing K2EDTA or LH at four different concentrations (1.76, 3.52, 7.04 and 17.6 mg/ml for K2EDTA; 16, 32, 64 and 160 IU/ml for LH) . Refractometric TP determination was performed on untreated and K2EDTA and LH aliquots with a hand-held refractometer and by spectophotometric Biuret method as the gold standard. Results Refractometric TP determination was overestimated in SF samples containing 10 times the recommended K2EDTA concentrations. Lower concentrations of K2EDTA and LH concentrations did not affect refractometric TP determinations. Main limitations Limited number of samples mostly obtained from large synovial structures. Conclusion To avoid incorrect TP determination, the use of LH containing collection tubes may be an appropriate alternative when the SF volume available is not enough to fill the K2EDTA collection tube.

The mapping of diattenuation, polarizance and retardance vector (normalized Stokes vector) on Poincare sphere, evaluated from Mueller matrix of optically anisotropic stromal region of cervical tissues, is presented for cervical precancer detection and its staging. This reveals that the changes in the polarization states shown by these normalized Stokes vectors corresponds to the degradation of linearly arranged collagen fibers, breakage of the collagen cross links in the stromal region and change in the density of scattering sites when cervical cancer evolves. The distinct nature of real and imaginary parts of the refractive index for linear, linear-45 and circular polarization from the optically anisotropic stromal region underscore the various polarization structures of the connective tissue region which get modified during the pathological changes. It has been found that versatility of these vectors for normal and precancerous cervical tissue of various grades may be utilized as a key distinction for qualitative staging of cervical precancer tissue. Quantitative classification of precancerous stages of cervical precancer has been determined with 95%-100% sensitivity and 93%-100% specificity through the evaluation of linear and circular diattenuation, linear polarizance and linear birefringence from the components of the respective vectors.

Background: High molecular weight (MW) synthetic colloids interfere with refractometry. The effect of low MW synthetic colloids on refractometry is unknown. Objectives: To investigate the effect of 6% hydroxyethyl starch 130/0.4 (HES) and 4% succinylated gelatin (GELO) on the refractometric measurement of total plasma protein (TPP) and colloid osmotic pressure (COP) in canine plasma. Methods: Heparinized plasma from 10 dogs was diluted with 0.9% saline (NaCl), HES, and GELO in ratios of plasma:fluid, 9:1, 8:2, 7:3, 6:4, and 5:5. The biuret TPP (TPPb), refractometric TPP (TPPr), and COP assays were performed for all dilutions. Agreement (bias, 95% limits of agreement (LOA)) was tested between the TPPb (NaCl) and the TPPr of the NaCl, HES, and GELO plasma dilutions, and between the TPPb (NaCl) and the TPPb of the HES and GELO plasma dilutions. Results: A small bias (0.06 g/dL, 95% LOA −0.51 to 0.39 g/dL) existed between TPPb (NaCl) and TPPr (NaCl). A large bias was found between TPPb (NaCl) and TPPr (HES) (1.38 g/dL, 95% LOA 0.30 to 3.10 g/dL), and TPPr (GELO) (1.18 g/dL, 95% LOA 0.53 to 2.88 g/dL). A small bias was found between TPPb (NaCl) and TPPb (HES) (0.08 g/dL, 95% LOA −2.04 to 2.20 g/dL), and a large bias was found between TPPb (NaCl) and TPPb (GELO) (1.00 g/dL, 95% LOA −0.59 to 2.60 g/dL). Increasing overestimation of TPPb (NaCl) by TPPr (HES and GELO) and TPPb (GELO) occurred with increasing colloid volumes. Conclusions: HES and GELO interfered with TPPr, and GELO also interfered with TPPb, in vitro. These variables should be interpreted with caution for dogs that have received these synthetic colloids.

BACKGROUND Inadequate absorption of colostral IgG1 is termed failure of transfer of passive immunity (FTPI). Dairy calves with FTPI have increased mortality and morbidity in their first 6 months of life. OBJECTIVES This study compared the clinical performance of 5 methods for diagnosing FTPI in Holstein calves. METHODS An observational study was performed using 160 Holstein heifer calves. Serum was harvested at 48 hours of age, and FTPI was assessed using a digital Brix refractometer for total solids measurements, and digital refractometry and the biuret method to measure serum total protein (STP) concentrations. Serum gamma-glutamyltransferase activity was measured with an automated analyzer, and serum IgG was measured with the zinc sulfate turbidity test and an enzyme-linked immunosorbent assay. Diagnostic test performance was compared with that of the reference method (FTPI defined as a serum total IgG concentration

PURPOSE: To evaluate the relationship between subjective and objective measurements of lens density and the energy of phacoemulsification.SETTING: University Eye Clinic, Maastricht University Medical Centre, Maastricht, the Netherlands.DESIGN: Cross-sectional study.METHODS: The study population included 69 patients. Nuclear opalescence (NO) was graded with the Lens Opacities Classification System III (LOCS III). Thereafter, lens density was measured objectively with Scheimpflug imaging, anterior segment optical coherent tomography and spectral fundus reflectometry (SFR). Cumulative dissipative energy (CDE) and total ultrasound time (US t.t.) of the phacoemulsification were noted. The relationship between the different measurement techniques and energy of phacoemulsification was assessed using Spearman's correlation coefficients.RESULTS: We found moderate to strong correlations between LOCS III and objective measurements of the lens density (rho's from 0.53 to 0.78, p < 0.05) and a moderate correlation between three objective measurement techniques (rho's between 0.29 and 0.57, p < 0.05). There was a moderate correlation between CDE, US t.t. and lens density measurements (rho's from 0.29 to 0.55, p < 0.05), and the highest correlation was found between CDE and NO scores. Exclusion of patients with advanced cortical and posterior subcapsular opacities improved the correlation between SFR and lens density measurements but not the correlation with the energy of phacoemulsification.CONCLUSION: Lens Opacities Classification System III has shown the highest correlation with phacoemulsification energy and may be a preferred technique for prediction of use of phacoemulsification energy. Advanced cortical and posterior opacities may interfere with the quality of objective measurements but do not affect the correlation between lens density measurements and phacoemulsification energy.

Background Transfer of passive immunity in calves can be assessed by direct measurement of immunoglobulin G (IgG) by methods such as radial immunodiffusion (RID) or turbidimetric immunoassay (TIA). IgG can also be measured indirectly by methods such as serum refractometry (REF) or Brix refractometry (BRIX). Objectives To determine the accuracy of REF and BRIX for assessment of inadequate transfer of passive immunity (ITPI) in calves. Design Systematic review and meta‐analysis of diagnostic accuracy studies. Methods Databases (PubMed and CAB Abstract, Searchable Proceedings of Animal Science) and Google Scholar were searched for relevant studies. Studies were eligible if the accuracy (sensitivity and specificity) of REF or BRIX was determined using direct measurement of IgG by RID or turbidimetry as the reference standard. The study population included calves

Micro-spectroscopy is a critical instrument for spectrum analysis in various applications such as chemical and biological analysis, environment detection, and hyperspectral imaging. However, current micro-spectral technique requires bulky and costly spectrometer. In this report, a new type of Bloch surface wave (BSW) based micro-spectrometer is proposed. A single silicon nanoparticle sitting on a dielectric multilayer substrate is used to excite the BSW which acts as a nanoscale unknown source. Taking advantage of the dispersion relations of BSWs, an abundant spectrally related database is formed that is useful for spectrum retrieval applications. Back-focal plane images are used to monitor the change of angular spectrum corresponding to the dispersion relationship of Bloch surface waves. Combined with an iterative algorithm, experimental retrieval of visible-range monochromatic and broadband light spectrums can be obtained. The resolution of the spectrometers can reach 2 nm across a wavelength range of 130 nm. The method in this work is CMOS compatible, enabling spectra retrieval for nanoscale radiators and can also be used to measure and retrieve the microscopic spectrum signal rapidly and timely without conventional scanning monochromator spectrometer.

Background Determination of serum total protein concentration is commonly performed by the biuret method. Refractometric measurement is a faster and less expensive alternative but its accuracy has not been determined in ruminants. Objective The purpose of the study was to compare the serum total protein concentrations in cattle, sheep, and goats measured by the biuret method with those obtained by refractometry. Methods Serum total protein concentration was determined in 120 cattle, 67 sheep, and 58 goat blood samples refractometrically and with the biuret method. The data were analyzed with a paired samples t-test, and Passing and Bablok regression equations and Bland and Altman plots were generated. Results There was a strong linear relationship between the total protein values determined with the refractometer and the biuret method in cattle, sheep, and goats. The statistical accuracy, which represents a bias correction factor that measures the deviation of the best-fit line from the 45° line through the origin, was 90.63% for cattle, 93.05% for sheep, and 91.76% for goats. The mean protein values determined with the refractometer were significantly lower than those measured with the biuret method in cattle and goats (P .05). Conclusions The evaluated refractometer was sufficiently accurate for the determination of serum total proteins in cattle, sheep, and goats, although it cannot be used interchangeably with the biuret method. The RIs should be corrected for negative bias based on the created equations.

PURPOSE Cardinal points are used for ray tracing through Gaussian systems. Anti-principal and anti-nodal points (which we shall refer to as the anti-cardinal points), along with the six familiar cardinal points, belong to a much larger set of special points. The purpose of this paper is to obtain a set of relationships and resulting equalities among the cardinal and anti-cardinal points and to illustrate them using Pascal's ring. METHODS The methodology used relies on Gaussian optics and the transference T. We make use of two equations, obtained via the transference, which give the locations of the six cardinal and four anti-cardinal points with respect to the system. We obtain equalities among the cardinal and anti-cardinal points. We utilise Pascal's ring to illustrate which points depend on frequency and their displacement with change in frequency. RESULTS Pascal described a memory schema in the shape of a hexagon for remembering equalities among the points and illustrating shifts in these points when an aspect of the system changes. We modify and extend Pascal's ring to include the anti-cardinal points. We make use of Pascal's ring extended to illustrate which points are dependent on the frequency of light and the direction of shift of the equalities with change in frequency. For the reduced eye the principal and nodal points are independent of frequency, but the focal points and the anti-cardinal points depend on frequency. For Le Grand's four-surface model eye all six cardinal and four anti-cardinal points depend on frequency. This has implications for definitions, particularly of chromatic aberrations of the eye, that make use of cardinal points and that themselves depend on frequency. CONCLUSIONS Pascal's ring and Pascal's ring extended are novel memory schema for remembering the equalities among the cardinal and anti-cardinal points. The rings are useful for illustrating changes among the equalities and direction of shift of points when an aspect of a system changes. Care should be taken when defining concepts that rely on cardinal points that depend on frequency.

The design of the smart plasmonic sensor, based on the periodical gold gratings grafted with PNIPAm was proposed and realized. Periodical gold structure was created on the polymer backing modified by excimer laser (1x2 cm2 modified area) and subsequently covered by thin gold film. In the next step 7 nm thick PNIPAm layer was grafted to gold surface through two step procedure: covalent attachment of 4-carboxybenzil groups followed by carboxyl groups activation and their coupling with amino-terminated PNIPAm. Surface properties of the prepared structure and their switchable behaviour were investigated using refractometry, wettability measurements, AFM in water medium, cyclic voltammetry, pH and temperature-dependent z-potential. The structure enables effective entrapping and SERS detection of azo dyes, significant reduction of minimal detectable concentration of target molecules and detection limit down to femtomolar concentrations. Excitation of plasmons on well ordered surface structure also shows excellent reproducibility of SERS system along the particular sample surface and between different samples too. Additionally, the reversible switching of wettability under repeated temperature changes on gold plasmonic platform allows multiple use of the developed system.

The eye is a very sophisticated system of optical elements for the preeminent sense of vision. In recent years, the number of laser surgery to correct the optical aberration such as myopia or astigmatism has significantly increased. Consequently, improving the knowledge related to the interactions of light with the eye is very important in order to enhance the efficiency of the surgery. For this reason, a complete optical characterization of the porcine eye is presented in this study. Kubelka-Munk and Inverse Adding-Doubling methods were applied to spectroscopy measurement to determine the absorption and scattering coefficients. Furthermore, the refractive index has been measured by ellipsometry. The different parameters were obtained for the cornea, lens, vitreous humor, sclera, iris, choroids and eyelid in the visible and infrared region. Thereafter, the results are implemented in a COMSOL Multiphysics® software to create an eye model. This model gives a better understanding of the propagation of light in the eye by adding optical parts such as the iris, the sclera or the ciliary bodies. Two simulations show the propagation of light from the cornea to the retina but also from the sclera to the retina. This last possibility provides a better understanding of light propagation during eye laser surgery such as, for example, transscleral cyclophotocoagulation. Figure: Eye simulation models allow the development of new laser treatments in a simple and safe way for patients. To this purpose, the creation of an eye simulated model based on optical parameters obtained from experimental data is presented in this study. This model will facilitate the understanding of the light propagation inside the porcine eye.

Introduction: Bobtail lizards (Tiliqua rugosa) are native to Australia. The only previous study on the hematology of this species documented just 6 animals. Objectives: The aims of this study were to characterize the light microscopy, ultrastructure and cytochemistry of blood cells, and evaluate CBCs of captive Bobtail lizards. Methods: Over 2 consecutive summers, heparinized venous blood was collected from the ventral coccygeal vein of 46 clinically healthy, captive indoor- or outdoor-housed adult Bobtails. Complete blood cell counts and smear evaluations were performed, and cytochemical stains and transmission electron microscopy were used to further characterize blood cells. Results: The eosinophils of this species were uniformly vacuolated: a unique feature not previously reported in reptiles. Heterophils were the predominant leukocyte, with fewer lymphocytes, azurophilic and nonazurophilic monocytes, occasional eosinophils, and basophils. Thrombocytes were frequently clumped. Slight polychromasia (0–15% of erythrocytes) was typically present. Hemogregarine parasites were seen on some smears. The range of CBC results was often wide. The PCV ranged from 11% to 38%. Total plasma proteins by refractometry were between 3.5 and 7.8 g/dL. Hemoglobin ranged between 2.6 and 12.6 g/dL by the modified hemoglobin-hydroxylamine method. Manual RBC count was 0.35–1.27 × 106/μL, and WBC count was 2.86–22.66 × 103/μL. Bobtail lizards housed outdoors had lower PCVs than indoor-housed animals. Bobtails with hemogregarine infections had lower PCVs than noninfected lizards. Conclusions: Ranges for CBC data were often very wide, influenced by preanalytic and analytic factors. Hemogregarine infection is associated with a decreased PCV, suggesting that some hemogregarine species are pathogenic in this population.

The formation of ultralong peptide crystalline nanobelts using a solvothermal approach from a di-phenylalanine gel within 10 min, where the self-assembly process is accelerated by several orders of magnitude compared with the month-long glutaraldehyde induction method previously reported, has been demonstrated. The solvothermal treatment can facilitate the phase separation of di-phenylalanine gels while speeding up the kinetics of the intramolecular cyclization reaction and concomitant crystallization. Moreover, the modulation effect of formaldehyde as an additive is revealed, and that a small amount of formaldehyde leads to thicker crystalline platelets capable of curved optical waveguiding that can potentially be applied in advanced bio-optical and optoelectronic devices, a rare feat with peptide-based crystals.

Common perception regards the nucleus as a densely packed object with higher refractive index (RI) and mass density than the surrounding cytoplasm. Here, the volume of isolated nuclei is systematically varied by electrostatic and osmotic conditions as well as drug treatments that modify chromatin conformation. The refractive index and dry mass of isolated nuclei is derived from quantitative phase measurements using digital holographic microscopy (DHM). Surprisingly, the cell nucleus is found to have a lower RI and mass density than the cytoplasm in four different cell lines and throughout the cell cycle. This result has important implications for conceptualizing light tissue interactions as well as biological processes in cells.

BACKGROUND The aim of this study was to analyse the influence of desalting temperature, fish thickness and desalting time on the mass transfer kinetics during the cod desalting process by physico-chemical analyses. RESULTS Both water uptake and salt loss increased with increasing temperature (15 °C > 10 °C > 5 °C) up to 24 h in ‘thicker’ pieces. The equilibrium achievement was faster in ‘thinner’ pieces and also with increasing temperature. Longer desalting times at 10 °C can be a good practice to be used during cod desalting at an industrial scale in order to obtain commercial products with higher yields. The faster mass transfer during desalting of ‘thinner’ pieces appears to follow three periods as a result of diffusion of the components (water, NaCl, and soluble proteins) because of the concentration differences, and pressure gradients due to expansion/shrinkage of the protein matrix, which is dependent on the NaCl content. The refractive index can be used by industry as an indirect measurement to determine the moment at which the ‘thicker’ samples are near the ZNaCl = YNaCl equilibrium. CONCLUSION Optimum combinations between the process variables analysed are essential in order to speed up the mass transfer kinetics during cod desalting at an industrial scale. © 2016 Society of Chemical Industry

The solid fat content (SFC) at different temperatures is an important characteristic of fat phases because it correlates to functionality in product applications. Consequently, this characteristic is also used to specify fat compositions in trade. Of three methods applicable, pulsed nuclear magnetic resonance (pMNR) is predominantly applied. Dilatometry and differential scanning calorimetry (DSC) find much less application. Handling with glass vials and high equipment costs make the search for alternatives to pNMR a useful endeavor. Optical refractometry is evaluated with respect to its potential to determine SFC values. Since refractometry is in the first place not suited for suspensions the positive results found are surprising. Applying temperature modulated optical refractometry (TMOR), isothermal optical refractometry with a superimposed temperature undulation yields repeatable results that are highly comparable to pNMR data. For the system studied (palm oil, coconut oil, partially hydrogenated palm oil), TMOR clearly outperforms DSC when pNMR is considered the method of reference. The key finding that refractive index is suitable to determine properties of suspensions is accompanied by the indications that refractometry has the potential to enable competitive methods within the fat technology. Practical Applications: The observation that refractometry can deliver quantitative data on fat suspensions enables the development of an array of new analytical methods. Next to SFC values and melting point, studies on the characterization of polymorphism can be envisioned. Since the device is robust and affordable, it could be in product development and quality control.

The authors demonstrate a form of two-photon-counting interferometry by measuring the coincidence counts between single-photon-counting detectors at an output port of a Mach-Zehnder Interferometer (MZI) following injection of broad-band time-frequency-entangled photon pairs (EPP) generated from collinear spontaneous parametric down conversion into a single input port. Spectroscopy and refractometry are performed on a sample inserted in one internal path of the MZI by scanning the other path in length, which acquires phase and amplitude information about the samples linear response. Phase modulation and lock-in detection are introduced to increase detection signal-to-noise ratio and implement a down-sampling technique for scanning the interferometer delay, which reduces the sampling requirements needed to reproduce fully the temporal interference pattern. The phase-modulation technique also allows the contributions of various quantum-state pathways leading to the final detection outcomes to be extracted individually. Feynman diagrams frequently used in the context of molecular spectroscopy are used to describe the interferences resulting from the coherence properties of time-frequency EPPs passing through the MZI. These results are an important step toward implementation of a proposed method for molecular spectroscopy, i.e. quantum-light-enhanced two-dimensional spectroscopy.

Chromophore-functionalized copoly(2-oxazoline)s are successfully evaluated as bottom antireflective coatings (BARCs) in high-resolution photolithography. With respect to UV light sources used in photolithographic production routines, anthracene is chosen as a chromophore. For application as polymer in BARCs, the copolymer poly(2-ethyl-2-oxazolin)45 -stat-poly(2-dec-9'-enyl-2-oxazolin)20 -stat-poly(2-(3'-(1"-(anthracen-9-ylmethyl)-1",2",3"-triazol-4-yl)propyl)-2-oxazolin)35 can be synthesized by the Huisgen cycloaddition click reaction of the copolymer poly(2-ethyl-2-oxazolin)45 -stat-poly(2-dec-9'-enyl-2-oxazolin)20 -stat-poly(2-pent-4'-inyl-2-oxazolin)35 and the corresponding azide-functionalized anthracenes. These copolymers can be crosslinked by the thermally induced thiol-ene reaction involving the unsaturated C=C bonds of the poly(2-dec-9'-enyl-2-oxazoline) repetition units and a multifunctional thiol as crosslinker. Tests of this BARC in a clean room under production conditions reveal a significant decrease of the swing-curve of a chemically amplified positive photoresist by more than 50%, hence significantly increasing the resolution of the photoresist.

Purpose Conjunctival compression observed in ultrahigh resolution optical coherence tomography (UHR-OCT) images of contact lens edges could be actual tissue alteration, may be an optical artefact arising from the difference between the refractive indexes of the lens material and the conjunctival tissue, or could be a combination of the two. The purpose of this study is to image the artefact with contact lenses on a non-biological (non-indentable) medium and to determine the origins of the observed conjunctival compression. Methods Two-dimensional cross-sectional images of the edges of a selection of marketed silicone hydrogel and hydrogel lenses (refractive index ranging from 1.40 to 1.43) were acquired with a research grade UHR-OCT system. The lenses were placed on three continuous surfaces, a glass sphere (refractive index n = 1.52), a rigid contact lens (n = 1.376) and the cornea of a healthy human subject (average n = 1.376). The displacement observed was analysed using ImageJ. Results The observed optical displacement ranged between 5.39(0.06) μm with Acuvue Advance and 11.99(0.18) μm with Air Optix Night & Day when the lens was imaged on the glass reference sphere. Similarly, on a rigid contact lens displacement ranged between 5.51(0.03) and 9.72(0.12) μm. Displacement was also observed when the lenses were imaged on the human conjunctiva and ranged from 6.49(0.80) μm for the 1-day Acuvue Moist to 17.4(0.22) μm for the Pure Vision contact lens. Conclusions An optical displacement artefact was observed when imaging a contact lens on two rigid continuous surfaces with UHR-OCT where compression or indentation of the surface could not have been a factor. Contact lenses imaged in situ also exhibited displacement at the intersection of the contact lens edge and the conjunctiva, likely a manifestation of both the artefact and compression of the conjunctiva.

CO2 capture is still a major challenge for the environment and decarbonization of gas streams. It has been industrially feasible for several decades using chemical absorption by alkanolamine solvents. The purpose of this work is to characterize the mass transfer in a spray contactor during CO2 absorption using monoethanolamine as a solvent. A new, effective way to investigate mass transfer is to measure the refractive index during the reactive absorption. Indeed, the concentration and temperature evolution inside the droplet during absorption induces a modification of the refractive index. By using rainbow refractometry technique (GRT), the liquid side mass transfer coefficient was measured in a controlled gas atmosphere during a transfer process in aqueous 30 % (w/w) monoethanolamine and compared to the literature.

In this paper seven salts of pyridinium-substituted tetraphenylethylene with different anions are reported. They show typical aggregation-induced emission. Crystal structures of three of the salts with (CF(3)SO(2))(2) N(-), CF(3) SO(3)(-), and SbF(6)(-) as the respective counter anions, are determined. The emission behavior of their amorphous and crystalline solids is investigated. Both amorphous and crystalline solids, except for the one with I(-), are highly emissive. Certain amorphous solids are red-emissive with almost the same quantum yields and fluorescence life-times. However, some crystalline solids are found to show different emission colors varying from green to yellow. Thus, their emission colors can be tuned by the counter anions. Furthermore, certain crystalline solids are highly emissive compared to the respective amorphous solids. Such solid-state emission behavior of these pyridinium-substituted tetraphenylethylene salts is interpreted on the basis of their crystal structures. In addition, optical waveguiding behavior of fabricated microrods is presented.

Background/aims The age-dependent changes in the optical reflection characteristics have been studied about skin hydration, melanin index, or skin color. However, the age-dependent changes in the optical reflection have little attention on inner skin structures. To control the factors affecting the optical reflection except for dermal matrix, subjects were selected as our guideline and we evaluated the optical reflection of subsurface on skin layers of two age groups. Method Young and old healthy volunteers were recruited after signing a written informed consent form. Facial skin was measured by means of noninvasive measurements: skin hydration, color, epidermis and dermis thickness, dermal density, subsurface reflectance, and transmittance. Result Compared to young group, old group showed that dermal density and thickness was decreased significantly although epidermis thickness was not changed. Conclusion In conclusion, dermal density is one of the major factors which affects the subsurface reflectance in skin.

Background The quantitative difference of the light reflected from a skin surface can be analyzed using parallel-polarized light (PPL) photography when combined with an analytic technique similar to colorimetric photography. Objective To improve the PPL photography technique as an assessment tool for the evaluation of skin condition using light-emitting diodes (LED) of different colors. Method Parallel-polarized light images were taken using white and green LED illuminators. The acquired images were transformed to CIELAB coordinates. An in-house skin conductance meter was constructed to assess skin hydration level. A dermatologist evaluated the clinical grading of dryness and glossiness. These clinical severities were also compared statistically with the CIELAB values. Results As with the green illuminator, the correlation analysis of whole sites showed that the L* value positively correlated with age (r = 0.18677, P

Background Bilirubin is stated to be a negative interferent in some biuret assays and thus could contribute to pseudohypoproteinemia in icteric samples. Objective The purpose of the study was to evaluate the magnitude of and reason for a falsely low total protein concentration in icteric serum when the protein concentration is measured with a bichromatic spectrophotometric biuret assay. Methods Commercially available bilirubin was dissolved in 0.1 M NaOH and mixed with sera from 2 dogs to achieve various bilirubin concentrations of up to 40 mg/dL (first set of samples) and 35 mg/dL (second set of samples, for confirmation of first set of results and to explore the interference). Biuret total protein and bilirubin concentrations were determined with a chemistry analyzer (Cobas 6000 with c501 module). Line graphs were drawn to illustrate the effects of increasing bilirubin concentrations on the total protein concentrations. Specific spectrophotometric absorbance readings were examined to identify the reason for the negative interference. Results High bilirubin concentrations created a negative interference in the Cobas biuret assay. The detectable interference occurred with a spiked bilirubin concentration of 10.7 mg/dL in one set of samples, 20.8 mg/dL in a second set. The interference was due to a greater secondary-absorbance reading at the second measuring point in the samples spiked with bilirubin, which possibly had converted to biliverdin. Conclusion Marked hyperbilirubinemia is associated with a falsely low serum total protein concentration when measured with a bichromatic spectrophotometric biuret assay. This can result in pseudohypoproteinemia and pseudohypoglobulinemia in icteric serum.

Background/purpose The speed of light (time of flight) into the skin is obviously relied to its structure, and might appear as a tool for non-invasive investigation of skin physico-chemical properties, among them aging is of primary importance. Though already published, such time of flight measurements have never been extensively correlated with other well-documented skin parameters such as localization, the influence of gender and age, the elasticity and roughness, and the water trans-epidermal diffusion (TEWL). Methods A specific practical device was designed to routinely measure the time of flight (TOF) of the light into the human skin ‘in vivo’, in a totally non-invasive process. This system was tested on volunteers, to relate the TOF parameter to the widely investigated skin properties already mentioned. An Infra-red laser at 1064 nm delivered powerful pulses of less than 1 ns duration, sent to the skin surface through a lossless fibre. The light backscattered at a given distance was collected and led onto an Avalanche Photodiode, and the mean TOF was measured on a fast sampling scope. A resolution and a reproducibility of a few picoseconds has been achieved. Experiments were carried out on 100 volunteers of both gender, aged from 18 to 65, on 12 different locations. Results No matter age and gender, important variations of TOF according to the localization were observed: On the inner forearm, an increase from wrist to elbow, and much higher values on the forehead and neck, whether orientation parallel or perpendicular to Langer lines did not appear significant. Ageing appeared to increase the TOF on forehead and neck, while this effect could not be confirmed on the forearm. Usual skin parameters such as elasticity, roughness and TEWL have been compared to TOF on the same location for each volunteer: TOF and skin roughness were significantly anti-correlated, i.e. the TOF got shorter when the Roughness increased, while a striking correlation was observed between TEWL and TOF. Conclusion These results assert the dependence of TOF on the nature of the skin upper layers (roughness, water diffusion) and on the dermis layer (ageing), and show the potential capabilities offered by TOF to investigate deeply into the skin structure. They have to be confirmed through further experiments, involving measurements at shorter wavelengths, at which the light path into the skin is much smaller, to get a distribution of the TOF inside the tissue.

Polymers for IR imaging: The preparation of high refractive index polymers (n = 1.75 to 1.86) via the inverse vulcanization of elemental sulfur is reported. High quality imaging in the near (1.5 μm) and mid-IR (3-5 μm) regions using high refractive index polymeric lenses from these sulfur materials was demonstrated.

Background The performance of a digital Atago PAL-USG Cat refractometer (Atago) was compared with a Schmidt and Haensch, Goldberg type refractometer (S+H). Materials and Methods Specific gravity of 47 canine and feline urine samples was determined with both refractometers and the results were compared with Passing–Bablok and Bland–Altman plots. In addition, the specific gravity of dilutions of 10% glucose, 10% NaCl, and 3% albumin solutions was determined and compared with expected values. Results Both refractometers consistently reported 1.000 with distilled water. The correlation between both refractometers based on Passing–Bablok plots of 47 urine samples was excellent (r = .99), but, in the Bland–Altman plots, there was a significant, proportional, negative error for the Atago readouts. This was also illustrated by the fact that 10 urine samples with an S+H result of > 1.030 were read out between 1.023 and 1.028 by Atago. Schmidt and Haensch results of various glucose solutions matched exactly expected values, but Atago results were lower. Likewise, S+H results with diluted NaCl solutions were closer to expected results than Atago results. In contrast, Atago results with dilutions of 3% albumin were closer to expected results than S+H results. Discussion The Atago refractometer reported lower specific gravity results in urine and standard solutions of glucose and NaCl, which could adversely affect clinical decisions concerning normal renal function based on solute concentrations determined in canine and feline urine samples.

Background and Objective The aim of this study was to obtain in vivo images of the microcirculation in tissues lining the gingival crevice in periodontally healthy volunteers and to assess the repeatability of the parameters measured. Material and Methods Video microscopy images of the microcirculation of tissues lining the gingival crevice were obtained from 20 periodontally healthy volunteers. Images were obtained with a single 1 mm diameter 1 pitch gradient index lens with a high numerical aperture and with a plain glass lens and illumination with a green 525 nm light-emitting diode and recorded using a video microscope. Results The morphological features of the vessels (including vessel diameter, vessel density, loops, branches, dilated vessels) were similar to those described previously in other mammals. The Kappa values for the assessment of morphology of the vessels using the gradient index lens range from 0.83 for branching to 0.91 for dilated and using the glass lens 0.47 for branching and 0.38 for dilated. Conclusions This novel system allowed for a consistent and repeatable assessment of the gingival microvasculature. However, there was some evidence of possible pressure artefacts in those cases where the measurements of separation between vessels exceeded 150 μm.

A powerful variation of traditional radical thiol-yne reaction with diphenylacetylene (DPA)-based starting materials leading to the quantitative and selective formation of the corresponding vinyl sulfides is reported. A variety of different thiols are shown to undergo reaction with DPA and the influence of their structure on reactivity is studied. The results obtained from the model reactions are then used to guide the efficient synthesis of hyperbranched poly(vinyl sulfide) (hb-PVS) systems by employing a dithiol and a trialkyne in an A2 + B3 approach. The polymers obtained show excellent solubility in common organic solvents and exhibit high refractive indices (e.g., 1.70 at 589 nm). The combined ease of processability and potential for cross-linking make these materials very interesting for applications, such as coatings for optical devices. The selective mono-addition thiol-yne reaction on DPA serves not only as a synthetic method for the preparation of PVS but could also be applied to the general modification of acetylene-containing materials.

Transparent, high water content (>65%), and cytocompatible hydrogels, which also possess super high refractive indices (RI > 1.5), are needed for ophthalmological applications. Most hydrogels can achieve either high RI or high water content but not both in the same system because water is a low RI material. Here, high water content/high RI hydrogels fabricated through elevated-temperature UV polymerization of an aqueous solution of acrylamide (AM) and methacrylamide (MAM) with tri(ethylene glycol) dimethacrylate (TEDA) crosslinker are reported. By varying the AM:MAM ratios (2:8 to 8:2) and crosslinker density (5 to 11 mol %), it is discovered that high water content (66%) AM:MAM copolymer hydrogels exhibiting anomalously high refractive indices (1.53); they are also colorless, transparent (99.4%), and cytocompatible with human keratinocytes.

Background Stretch marks or striae distensae (SD) are characterized by epidermal atrophy following repeated over-stretching of the skin tissue. The objective of this study was to investigate the skin texture and microstructure of SD lesions compared to those of normal adjacent skin in vivo using non-invasive methods. Methods A population of 26 women and 3 men with SD were examined after giving written informed consent. Following clinical grading, skin replicas were collected, confocal microscopy was performed on SD lesions and healthy neighboring skin. Skin surface texture parameters were calculated using 3D image analysis of the skin replicas and epidermal and dermal microstructure were evaluated by analysis of the confocal images. In a parallel study, histological analysis was performed on 6 skin biopsies taken from abdominal reduction surgeries in areas where skin exhibited SD. Results Analysis of the skin surface texture showed that the SD area was more anisotropic and with higher skin roughness than the adjacent skin in terms of directions of skin microglyphics. Confocal microscopy demonstrated that SD were characterized by shallower dermal papillae (P

Exceptional nonlinear optical (NLO) switching behavior, including an extremely large contrast (on/off) of ∼35 and high NLO coefficients, is displayed by a solid-state reversible quadratic NLO switch. The favorable results, induced by very fast molecular motion and anionic ordering, provides impetus for the design of a novel second-harmonic-generation switch involving molecular motion.

A quick recovery: A semitransparent composite conductor comprising a layer of silver nanowire percolation network inlaid in the surface layer of a Diels-Alder-based healable polymer film is fabricated. The composite is flexible and highly conductive, and is capable of both structural and electrical healing via heating. Cut samples that completely lose their conductivity can recover 97% of it within 5 minutes of heating at 110 °C. The cutting and healing can be repeated at the same location for multiple cycles.

The self-organization of ferroelectric columnar liquid crystals (FCLCs) is demonstrated. Columnar order is spontaneously formed in thin films made by the wet-process due to its liquid crystallinity. Electric-field application results in high optical quality and uniform spontaneous polarization. Such good processability and controllability of the wet-processed FCLC films provide us with potential organic ferroelectric materials for device applications.