MiniRead: A simple and inexpensive do-it-yourself device for multiple analyses of micro-organism growth kinetics.

Fitness in micro-organisms can be proxied by growth parameters on different media and/or temperatures. This is achieved by measuring optical density at 600 nm using a spectrophotometer, which measures the effect of absorbance and side scattering due to turbidity of cells suspensions. However, when growth kinetics must be monitored in many 96-well plates at the same time, buying several 96-channel spectrophotometers is often beyond budgets. The MiniRead device presented here is a simple and inexpensive do-it-yourself 96-well temperature-controlled turbidimeter designed to measure the interception of white light via absorption or side scattering through liquid culture medium. Turbidity is automatically recorded in each well at regular time intervals for up to several days or weeks. Output tabulated text files are recorded into a micro-SD memory card to be easily transferred to a computer. We propose also an R package which allows (1) to compute the nonlinear calibration curves required to convert raw readings into cell concentration values, and (2) to analyze growth kinetics output files to automatically estimate proxies of growth parameters such as lag time, maximum growth rate, or cell concentration at the plateau.

Complex coacervation of pea albumin-pectin and ovalbumin-pectin assessed by isothermal titration calorimeter and turbidimetry.

BACKGROUND: This study investigates the complexation of a pea albumin-rich fraction and ovalbumin with pectin of different degrees of esterification (DE) and blockiness (DB) as a function of pH and biopolymer mixing ratio by turbidimetric titration and isothermal titration calorimetry (ITC). RESULTS: Turbidimetric analysis found maximum complexation occurred at a mixing ratio of 4:1 for pea albumin with high methoxy pectin, 8:1 for pea albumin with low methoxy pectin, and 8:1 for ovalbumin with low methoxy pectin. In the case of ovalbumin with high methoxy pectin, interactions were very weak. The pectin with high levels of esterification and blockiness displayed greater interactions with the pea albumin in both turbidimetry and ITC. However, low methoxy pectin imparted better interactions with ovalbumin and displayed higher optical density values than high methoxy pectin. CONCLUSIONS: The current study indicated that the different thermodynamic parameters of PA-pectin complexes can be tuned by controlling the structural characteristics (DB, DE, and d-galacturonic acid) of the pectin. © 2020 Society of Chemical Industry.

Use of High-Resolution Pressure Nephelometry To Measure Gas Vesicle Collapse as a Means of Determining Growth and Turgor Changes in Planktonic Cyanobacteria.

Previous work has demonstrated that the physical properties of intracellular bacterial gas vesicles (GVs) can be analyzed in vivo using pressure nephelometry. In analyzing the buoyant state of GV-containing cyanobacteria, hydrostatic pressure within a sample cell is increased in a stepwise manner, where the concomitant collapse of GVs due to pressure and the resultant decrease in suspended cells are detected by changes in nephelometric scattering. As the relative pressure at which GVs collapse is a function of turgor pressure and cellular osmotic gradients, pressure nephelometry is a powerful tool for assaying changes in metabolism that affect turgor, such as photosynthetic and osmoregulatory processes. We have developed an updated and automated pressure nephelometer that utilizes visible-infrared (Vis-IR) spectra to accurately quantify GV critical collapse pressure, critical collapse pressure distribution, and cell turgor pressure. Here, using the updated pressure nephelometer and axenic cultures of Microcystis aeruginosa PCC7806, we demonstrate that GV critical collapse pressure is stable during mid-exponential growth phase, introduce pressure-sensitive turbidity as a robust metric for the abundance of gas-vacuolate cyanobacteria, and demonstrate that pressure-sensitive turbidity is a more accurate proxy for abundance and growth than photopigment fluorescence. As cyanobacterium-dominated harmful algal bloom (cyanoHAB) formation is dependent on the constituent cells possessing gas vesicles, characterization of environmental cyanobacteria populations via pressure nephelometry is identified as an underutilized monitoring method. Applications of this instrument focus on physiological and ecological studies of cyanobacteria, for example, cyanoHAB dynamics and the drivers associated with cyanotoxin production in aquatic ecosystems.IMPORTANCE The increased prevalence of bloom-forming cyanobacteria and associated risk of exposure to cyanobacterial toxins through drinking water utilities and recreational waterways are growing public health concerns. Cost-effective, early-detection methodologies specific to cyanobacteria are crucial for mitigating these risks, with a gas vesicle-specific signal offering a number of benefits over photopigment fluorescence, including improved detection limits and discrimination against non-gas-vacuolate phototrophs. Here, we present a multiplexed instrument capable of quantifying the relative abundance of cyanobacteria based on the signal generated from the presence of intracellular gas vesicles specific to bloom-forming cyanobacteria. Additionally, as cell turgor can be measured in vivo via pressure nephelometry, the measurement furnishes information about the internal osmotic pressure of gas-vacuolate cyanobacteria, which relates to the metabolic state of the cell. Together these advances may improve routine waterway monitoring and the mitigation of human health threats due to cyanobacterial blooms.

Evaluation of the Alfred™ turbidity monitoring system (Alifax®) following sonication in the diagnosis of central venous catheter colonization.

The conventional diagnostic techniques for catheter colonization (CC) take at least 48 h to yield results. Therefore, new diagnostic procedures that speed up the time necessary for results are needed. Our main objective was to assess the efficacy of the combination of sonication, turbidity monitoring, and MALDI-TOF to detect CC and catheter-related bloodstream infection (C-RBSI). For 1 year, we assessed central venous catheter (CVC) tips that arrived at the microbiology laboratory from adult patients admitted to our institution. CVC tips were cut, inoculated into 2.5 ml of BHI, and sonicated for 1 min. The suspension was then processed using Gram stain, quantitative culture (gold standard), and preincubation on the Alfred™ system. We analyzed the validity values of our new diagnostic approach for prediction of CC and C-RBSI and compared them with those of the gold standard. We collected a total of 167 catheters, 33 (19.8%) of which were colonized. We confirmed 21 episodes of C-RBSI. The distribution of microorganisms in colonized CVCs was as follows: Gram-positive, 68.4%; Gram-negative, 5.3%; and yeasts, 26.3%. The validity values for CC and C-RBSI using the new procedure were as follows: S, 39.4%/61.9%; Sp, 100%/100%; PPV, 100%/100%; and NPV, 87.0%/94.8%. The combination of sonication with a pre-incubation period based on turbidity monitoring using the Alfred™ system followed by MALDI-TOF proved to be a useful tool that was faster than conventional culture for ruling out C-RBSI. Future studies are needed to assess the clinical and economic impact of this diagnostic approach.

The Observation and Analysis of Internal Quality Control of Cystatin C in China from 2014 to 2017.

BACKGROUND: This study observed and analyzed the internal quality control (IQC) of cystatin C (CysC) so that we can have overall knowledge about imprecision levels in Chinese medical laboratories. METHODS: Using the software developed by the National Center for Clinical Laboratories (NCCL), we can get the IQC information of CysC from 2014 to 2017. Then the proportion of laboratories meeting five quality specifications (pass rates) were calculated and the current CVs (coefficient of variation) were also compared among subgroups and years. RESULTS: We find that the current CVs between 2014 and 2017 show significant differences (p = 0.016) and the proportion of laboratories meeting the 1/3 TEa specification distributes randomly from 2014 to 2017 but all of them exceed 80 percent. When the optimum specification is applied, the pass rates all become very low and the distributions are wide spread (from 3.63% to 6.74%). Beckman, Roche, and Hitachi are mainstream analyzers, making up as much as 78% to 85% of all. We can see a significant difference of the pass rate between Beckman and Hitachi in 2014 (p = 0.005). The primary means of detecting CysC is particle-enhanced turbidimetric immunoassay (PETIA) which makes up 30.57% - 32.64% of detection methods. The good news is that the IQC practice has improved greatly from 2014 to 2017 in laboratories in China. CONCLUSIONS: Medical laboratories have made some progress in IQC from 2014 to 2017, but it is still not satisfactory. As a result, there is a long way to go to improve detection quality of laboratories in China.

Performance evaluation of serum IgG subclass quantification using a SPAPLUS turbidimetric analyzer and comparison with the BNII nephelometer.

IgG consists of four subclasses: IgG1, IgG2, IgG3, and IgG4. Changes in the serum concentration of each subclass reflect different clinical situations, and quantification of each subclass is important to assess patients' clinical states. Herein, we evaluated the analytical performance of the SPAPLUS turbidimetric analyzer (The Binding Site, Birmingham, UK) for IgG subclass. Precision, linearity, comparison with the BNII system (Siemens Healthineers, Erlangen, Germany), and reference interval were assessed according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. The repeatability and within-laboratory precision were within 5% for all IgG subclasses. The coefficient of determination (R2) was higher than 0.99 for the analytical measurement range in all IgG subclasses. Comparison between SPAPLUS and BNII revealed significant differences in IgG1, IgG3, and IgG4 (p < .0001). IgG1 and IgG4 values were lower in SPAPLUS than BNII. On the other hand, IgG3 values were higher in SPAPLUS than BNII. The SPAPLUS turbidimetric analyzer exhibited good analytical performance for quantification of four IgG subclasses. Because of the differences between SPAPLUS and BNII, follow-up test for disease monitoring should be performed with same instrument.

Factors related to Secchi depths and their stability over time as determined from a probability sample of US lakes.

A probabilistic sample of lakes in the 48 coterminous US lakes was made by the United States Environmental Protection Agency in the 2007 National Lakes Assessment. Because of the statistical design, the results of our analyses of Secchi depths (SD) apply to a population of 45,265 lakes. We found statistically significant differences in mean Secchi depths between natural (1.57 m) and man-made lakes (1.18 m). The most important variable correlated with SD was turbidity, an optical measure related to suspended particles in the water column. For most lakes, chlorophyll a was highly correlated with both turbidity and SD, but several lakes had more turbidity and lower SD than expected based on chlorophyll a alone, indicating that non-algal suspended solids were an important factor. On an ecoregion basis, the non-algal suspended solids in the lake waters were related to the average levels of suspended solids in streams located in that ecoregion, and the non-algal suspended solids were more important in man-made than natural lakes. Phosphorus and nitrogen were directly correlated with chlorophyll a and turbidity and inversely correlated with SD. Based on diatom-inferred Secchi depths for the tops and bottoms of sediment cores from lakes in Ecoregions VIII and VII (excluding lakes in Minnesota) representing 40% of the natural lakes in the US, there has been no decrease in water transparency in that population of lakes in the past 70 or more years when the US population increased by 134%. We do not have information to determine if the other 60% of lakes have or have not changed.

Interferometric Near-Infrared Spectroscopy (iNIRS) for determination of optical and dynamical properties of turbid media.

We introduce and implement interferometric near-infrared spectroscopy (iNIRS), which simultaneously extracts optical and dynamical properties of turbid media through analysis of a spectral interference fringe pattern. The spectral interference fringe pattern is measured using a Mach-Zehnder interferometer with a frequency-swept narrow linewidth laser. Fourier analysis of the detected signal is used to determine time-of-flight (TOF)-resolved intensity, which is then analyzed over time to yield TOF-resolved intensity autocorrelations. This approach enables quantification of optical properties, which is not possible in conventional, continuous-wave near-infrared spectroscopy (NIRS). Furthermore, iNIRS quantifies scatterer motion based on TOF-resolved autocorrelations, which is a feature inaccessible by well-established diffuse correlation spectroscopy (DCS) techniques. We prove this by determining TOF-resolved intensity and temporal autocorrelations for light transmitted through diffusive fluid phantoms with optical thicknesses of up to 55 reduced mean free paths (approximately 120 scattering events). The TOF-resolved intensity is used to determine optical properties with time-resolved diffusion theory, while the TOF-resolved intensity autocorrelations are used to determine dynamics with diffusing wave spectroscopy. iNIRS advances the capabilities of diffuse optical methods and is suitable for in vivo tissue characterization. Moreover, iNIRS combines NIRS and DCS capabilities into a single modality.

The Roche Immunoturbidimetric Albumin Method on Cobas c 501 Gives Higher Values Than the Abbott and Roche BCP Methods When Analyzing Patient Plasma Samples.

BACKGROUND: Serum/plasma albumin is an important and widely used laboratory marker and it is important that we measure albumin correctly without bias. We had indications that the immunoturbidimetric method on Cobas c 501 and the bromocresol purple (BCP) method on Architect 16000 differed, so we decided to study these methods more closely. METHOD: A total of 1,951 patient requests with albumin measured with both the Architect BCP and Cobas immunoturbidimetric methods were extracted from the laboratory system. A comparison with fresh plasma samples was also performed that included immunoturbidimetric and BCP methods on Cobas c 501 and analysis of the international protein calibrator ERM-DA470k/IFCC. RESULTS: The median difference between the Abbott BCP and Roche immunoturbidimetric methods was 3.3 g/l and the Roche method overestimated ERM-DA470k/IFCC by 2.2 g/l. The Roche immunoturbidimetric method gave higher values than the Roche BCP method: y = 1.111x - 0.739, R² = 0.971. CONCLUSION: The Roche immunoturbidimetric albumin method gives clearly higher values than the Abbott and Roche BCP methods when analyzing fresh patient samples. The differences between the two methods were similar at normal and low albumin levels.

Measuring Subvisible Particles in Protein Formulations Using a Modified Light Obscuration Sensor with Improved Detection Capabilities.

Although light obscuration is the "gold standard" for subvisible particle measurements in biopharmaceutical products, the current technology has limitations with respect to the detection of translucent proteinaceous particles and particles of sizes smaller and around 2 µm. Here, we describe the evaluation of a modified light obscuration sensor utilizing a novel measuring mode. Whereas standard light obscuration methodology monitors the height (amplitude) of the signal, the new approach monitors its length (width). Experimental evaluation demonstrated that this new detection mode leads to improved detection of subvisible particles of sizes smaller than 2 µm, reduction of artifacts during measurements especially of low concentrations of translucent protein particles, and higher counting accuracy as compared to flow imaging microscopy and standard light obscuration measurements.

Linear polarization optimized Stokes polarimeter based on four-quadrant detector.

A four-quadrant detector (4QD) consists of four well-balanced detectors. We report on a Stokes polarimeter with optimal linear polarization measurements based on a 4QD. We turned the four intensity-detection channels into four polarization-analyzing channels by placing four polarizers and one quarter-wave plate in front of the individual detectors. An optimization method for the four polarization-analyzing channels is proposed to improve measurement accuracy. Considering applications in favor of linear polarization measurements instead of global optimization for all the possible states of polarization (SOP), we optimize the polarimeter first for the linear polarization components and then for the circular polarization component. The polarimeter is capable of simultaneous measurements of fast varying SOP with improved performance for the linear polarizations.

Binocular open-view system to perform estimations of aberrations and scattering in the human eye.

We present a system that integrates a double-pass (DP) instrument and a Hartmann-Shack (HS) wavefront sensor to provide information not only on aberrations, but also on the scattering that occurs in the human eye. A binocular open-view design permits evaluations to be made under normal viewing conditions. Furthermore, the system is able to compensate for both the spherical and astigmatic refractive errors that occur during measurements by using devices with configurable optical power. The DP and HS techniques provide comparable data after estimating wavefront slopes with respect to the intersections of an ideal grid and compensating for residual errors caused by the optical defects of the measuring system. Once comparable data is obtained, it is possible to use this combined manner of assessment to provide information on scattering. Measurements in an artificial eye suggest that the characteristics of the ocular fundus may induce deviations of DP with respect to the HS data. These differences were quantified in terms of the modulation transfer function in young, healthy eyes measured in infrared light to demonstrate the potential use of the system in visual optics studies.

Airborne system for multispectral, multiangle polarimetric imaging.

In this paper, we describe the design, fabrication, calibration, and deployment of an airborne multispectral polarimetric imager. The motivation for the development of this instrument was to explore its ability to provide information about water constituents, such as particle size and type. The instrument is based on four 16 MP cameras and uses wire grid polarizers (aligned at 0°, 45°, 90°, and 135°) to provide the separation of the polarization states. A five-position filter wheel provides for four narrow-band spectral filters (435, 550, 625, and 750 nm) and one blocked position for dark-level measurements. When flown, the instrument is mounted on a programmable stage that provides control of the view angles. View angles that range to ±65° from the nadir have been used. Data processing provides a measure of the polarimetric signature as a function of both the view zenith and view azimuth angles. As a validation of our initial results, we compare our measurements, over water, with the output of a Monte Carlo code, both of which show neutral points off the principle plane. The locations of the calculated and measured neutral points are compared. The random error level in the measured degree of linear polarization (8% at 435) is shown to be better than 0.25%.

Label-free biomarker assay in a microresistive pulse sensor via immunoaggregation.

We present a label-free biomarker detection method based on immunoaggregation and resistive pulse sensing technology. In this approach, target biomarkers and antibody (Ab)-functionalized microparticles are mixed to form biomarker-microparticle aggregates. A resistive pulse sensor is then used to measure the sizes and count the number of aggregates. The measured volume fraction of the aggregates represents the concentration of the targeted biomarker. In our tests, human ferritin, used as a biomarker, triggered the aggregation of antiferritin Ab-functionalized microparticles in phosphate-buffered saline (PBS). The volume fraction of aggregates increased with the increased ferritin concentration. We also demonstrated the detection of human ferritin in 10% fetal bovine serum (FBS) to mimic a real detection environment in complex media. The detection range from 0.1 to 208 ng/mL was achieved. In addition, we demonstrated that the detection range can be shifted to lower and higher concentrations by decreasing and increasing microparticle concentrations. This biomarker detection method is label-free, rapid, and able to quantitatively measure the concentration of any macromolecular biomarker as long as an antibody can be found, with simple measurement setup and sample preparations.

Propagation factors of cosine-Gaussian-correlated Schell-model beams in non-Kolmogorov turbulence.

Based on the extended Huygens-Fresnel principle and second-order moments of the Wigner distribution function (WDF), we have studied the relative root-mean-square (rms) angular width and the propagation factor of cosine-Gaussian-correlated Schell-model (CGSM) beams propagating in non-Kolmogorov turbulence. It has been found that the CGSM beam has advantage over the Gaussian Schell-model (GSM) beam for reducing the turbulence-induced degradation, and this advantage will be more obvious for the beams with larger parameter n and spatial coherence δ or under the condition of stronger fluctuation of turbulence. The CGSM beam with larger parameter n or smaller spatial coherence δ will be less affected by the turbulence. In addition, the effects of the slope-parameter α, inner and outer scale and the refractive-index structure constant of the non-Kolmogorov's power spectrum on the propagation factor are also analyzed in detailed.

Electromagnetic sinc Schell-model beams and their statistical properties.

A class of electromagnetic sources with sinc Schell-model correlations is introduced. The conditions on source parameters guaranteeing that the source generates a physical beam are derived. The evolution behaviors of statistical properties for the electromagnetic stochastic beams generated by this new source on propagating in free space and in atmosphere turbulence are investigated with the help of the weighted superposition method and by numerical simulations. It is demonstrated that the intensity distributions of such beams exhibit unique features on propagating in free space and produce a double-layer flat-top profile of being shape-invariant in the far field. This feature makes this new beam particularly suitable for some special laser processing applications. The influences of the atmosphere turbulence with a non-Kolmogorov power spectrum on statistical properties of the new beams are analyzed in detail.

Optical parametrically gated microscopy in scattering media.

High-resolution imaging in turbid media has been limited by the intrinsic compromise between the gating efficiency (removal of multiply-scattered light background) and signal strength in the existing optical gating techniques. This leads to shallow depths due to the weak ballistic signal, and/or degraded resolution due to the strong multiply-scattering background--the well-known trade-off between resolution and imaging depth in scattering samples. In this work, we employ a nonlinear optics based optical parametric amplifier (OPA) to address this challenge. We demonstrate that both the imaging depth and the spatial resolution in turbid media can be enhanced simultaneously by the OPA, which provides a high level of signal gain as well as an inherent nonlinear optical gate. This technology shifts the nonlinear interaction to an optical crystal placed in the detection arm (image plane), rather than in the sample, which can be used to exploit the benefits given by the high-order parametric process and the use of an intense laser field. The coherent process makes the OPA potentially useful as a general-purpose optical amplifier applicable to a wide range of optical imaging techniques.

Imaging in scattering media using correlation image sensors and sparse convolutional coding.

Correlation image sensors have recently become popular low-cost devices for time-of-flight, or range cameras. They usually operate under the assumption of a single light path contributing to each pixel. We show that a more thorough analysis of the sensor data from correlation sensors can be used can be used to analyze the light transport in much more complex environments, including applications for imaging through scattering and turbid media. The key of our method is a new convolutional sparse coding approach for recovering transient (light-in-flight) images from correlation image sensors. This approach is enabled by an analysis of sparsity in complex transient images, and the derivation of a new physically-motivated model for transient images with drastically improved sparsity.

POLVSM (Polarized Volume Scattering Meter) instrument: an innovative device to measure the directional and polarized scattering properties of hydrosols.

An innovative instrument dedicated to the multispectral measurements of the directional and polarized scattering properties of the hydrosols, so-called POLVSM, is described. The instrument could be used onboard a ship, as a benchtop instrument, or at laboratory. The originality of the POLVSM concept relies on the use of a double periscopic optical system whose role is (i) to separate the plane containing the light source from the scattering plane containing the sample and the receiver and (ii) to prevent from any specularly reflected light within the sample chamber. As a result, a wide range of scattering angle, namely from 1° to 179°, is covered by the detector. Another originality of the instrument is to measure the Mueller scattering matrix elements, including the degree of polarization. A relevant calibration procedure, which could be of great interest as well for other instruments, is proposed to convert the raw data into physical units. The relative uncertainty in POLVSM data was determined at ± 4.3%. The analysis of measurements of the volume scattering function and degree of polarization performed under controlled conditions for samples dominated either by inorganic hydrosols or phytoplankton monospecific species showed a good consistency with literature, thus confirming the good performance of the POLVSM device. Comparisons of POLVSM data with theoretical calculations showed that Mie theory could reproduce efficiently the measurements of the VSF and degree of polarization for the case of inorganic hydrosols sample, despite the likely non sphericity of these particles as revealed by one of the element of the Mueller matrix. Our results suggested as well that a sophisticated modeling of the heterogeneous internal structure of living cells, or at least, the use of layered sphere models, is needed to correctly predict the directional and polarized effects of phytoplankton on the oceanic radiation. The relevance of performing angularly resolved measurements of the Mueller scattering elements to gain understanding on the mechanisms processes involved in the scattering of light by marine particles, which has important implications for ocean color remote sensing studies, is demonstrated.