Raman Spectroscopy of Disperse Systems with Varying Particle Sizes and Correction of Signal Losses.

In this paper, a dispersion of glass beads of different sizes in an ammonium nitrate solution is investigated with the aid of Raman spectroscopy. The signal losses caused by the dispersion are quantified by an additional scattered light measurement and used to correct the measured ammonium nitrate concentration. Each individual glass bead represents an interface at which the excitation laser is deflected from its direction causing distortion in the received Raman signal. It is shown that the scattering losses measured with the scattered light probe correlate with the loss of the Raman signal, which means that the data obtained can be used to correct the measured values. The resulting correction function considers different particle sizes in the range of 2-99 µm as well as ammonium nitrate concentrations of 0-20 wt% and delivers an RMSEP of 1.952 wt%. This correction provides easier process access to dispersions that were previously difficult or impossible to measure.

Raman Spectroscopy of Glass Beads in Ammonium Nitrate Solution and Compensation of Signal Losses.

In the present study, the influence of disperse systems on Raman scattering was investigated. How an increasing particle concentration weakens the quantitative signal of the Raman spectrum is shown. Furthermore, the change in the position of the optimal measurement point in the fluid was considered in detail. Additional transmission measurements can be used to derive a simple and robust correction method that allows the actual concentration of the continuous phase to be determined with a standard deviation of 2.6%.

Liquid Mixing on Falling Films: Marker-Free, Molecule-Sensitive 3D Mapping Using Raman Imaging.

Following up on a proof of concept, this publication presents a new method for mixing mapping on falling liquid films. On falling liquid films, different surfaces, plain or structured, are common. Regarding mixing of different components, the surface has a significant effect on its capabilities and performance. The presented approach combines marker-free and molecule-sensitive measurements with cross-section mapping to emphasize the mixing capabilities of different surfaces. As an example of the mixing capabilities on falling films, the mixing of sodium sulfate with tap water is presented, followed by a comparison between a plain surface and a pillow plate. The method relies upon point-by-point Raman imaging with a custom-built high-working-distance, low-depth-of-focus probe. To compensate for the long-time measurements, the continuous plant is in its steady state, which means the local mixing state is constant, and the differences are based on the liquids' position on the falling film, not on time. Starting with two separate streams, the mixing progresses by falling down the surface. In conclusion, Raman imaging is capable of monitoring mixing without any film disturbance and provides detailed information on liquid flow in falling films.

Spatial Omics Imaging of Fresh-Frozen Tissue and Routine FFPE Histopathology of a Single Cancer Needle Core Biopsy: A Freezing Device and Multimodal Workflow.

The complex molecular alterations that underlie cancer pathophysiology are studied in depth with omics methods using bulk tissue extracts. For spatially resolved tissue diagnostics using needle biopsy cores, however, histopathological analysis using stained FFPE tissue and the immunohistochemistry (IHC) of a few marker proteins is currently the main clinical focus. Today, spatial omics imaging using MSI or IRI is an emerging diagnostic technology for the identification and classification of various cancer types. However, to conserve tissue-specific metabolomic states, fast, reliable, and precise methods for the preparation of fresh-frozen (FF) tissue sections are crucial. Such methods are often incompatible with clinical practice, since spatial metabolomics and the routine histopathology of needle biopsies currently require two biopsies for FF and FFPE sampling, respectively. Therefore, we developed a device and corresponding laboratory and computational workflows for the multimodal spatial omics analysis of fresh-frozen, longitudinally sectioned needle biopsies to accompany standard FFPE histopathology of the same biopsy core. As a proof-of-concept, we analyzed surgical human liver cancer specimens using IRI and MSI with precise co-registration and, following FFPE processing, by sequential clinical pathology analysis of the same biopsy core. This workflow allowed for a spatial comparison between different spectral profiles and alterations in tissue histology, as well as a direct comparison for histological diagnosis without the need for an extra biopsy.

Establishing Reliable Research Data Management by Integrating Measurement Devices Utilizing Intelligent Digital Twins.

One of the main topics within research activities is the management of research data. Large amounts of data acquired by heterogeneous scientific devices, sensor systems, measuring equipment, and experimental setups have to be processed and ideally be managed by Findable, Accessible, Interoperable, and Reusable (FAIR) data management approaches in order to preserve their intrinsic value to researchers throughout the entire data lifecycle. The symbiosis of heterogeneous measuring devices, FAIR principles, and digital twin technologies is considered to be ideally suited to realize the foundation of reliable, sustainable, and open research data management. This paper contributes a novel architectural approach for gathering and managing research data aligned with the FAIR principles. A reference implementation as well as a subsequent proof of concept is given, leveraging the utilization of digital twins to overcome common data management issues at equipment-intense research institutes. To facilitate implementation, a top-level knowledge graph has been developed to convey metadata from research devices along with the produced data. In addition, a reactive digital twin implementation of a specific measurement device was devised to facilitate reconfigurability and minimized design effort.

Film Thickness and Glycerol Concentration Mapping of Falling Films Based on Fluorescence and Near-Infrared Technique.

Falling film evaporation processes involve high fluid velocities with continuous variations in local film thickness, fluid composition, and viscosity. This contribution presents a parallel and complementary film thickness and concentration mapping distribution in falling films using a non-invasive fluorescence and near-infrared imaging technique. The experiments were performed with a mixture of glycerol/water with a mass fraction from 0 to 0.65 gglycgtotal-1 and operating ranges similar to evaporation processes. The measurement system was designed by integrating two optical measurement methods for experimental image analysis. The film thickness was evaluated using a VIS camera and high-power LEDs at 470 nm. The local glycerol concentration gglycgtotal-1 was determined using a NIR camera and high-power LEDs at 1050, 1300, 1450 and 1550 nm. A multiwavelength analysis with all NIR wavelengths was implemented with a better correlation for falling films at low flow velocity. The results show an improvement in the analysis of falling films with high flow velocities up to almost 500 mm/s by using only the 1450 nm wavelength and the fluorescence measurement. Simultaneous imaging analysis of film thickness and concentration in falling films provides further insight into understanding mass and heat transport and thus supports the optimization of falling film evaporators.

New Conceptional Study of a Portable Highly Sensitive Photometric Raman Sensor.

Quality control and reaction monitoring are necessary to ensure the consistency of any kind of mixing or reaction process. In this context, a novel portable high-sensitivity sensor prototype based on the Raman effect is presented in this study. The elongated probe head is designed for (but not limited to) monitoring high temperature batch processes for quality assurance. Thanks to the highly sensitive special detectors, concentration differences of up to 50 mmol/L can currently be detected, which facilitates compliance with high product quality standards. In addition, seamless reaction tracking is possible. Small individual adjustments through simple, intuitive mechanical components provide a high degree of flexibility in reaction selection by the end user. Specially developed software automates the evaluation process and gives the user visual signals about the current status and progress of the batch as well as an emergency stop if temperature limits could damage individual components. By using all the individual components developed, the problem of the limited integration times of previous spectrometric measuring instruments could be reduced. The prototype was validated using concentration measurements of various substances that occur as standard in batch processes. In addition, this article provides an outlook on the fact that Raman measurements can also be carried out successfully and reliably in explosive environments in the future with the prototype presented.

Marker-Free, Molecule Sensitive Mapping of Disturbed Falling Fluid Films Using Raman Imaging.

Technical liquid flow films are the basic arrangement for gas fluid transitions of all kinds and are the basis of many chemical processes, such as columns, evaporators, dryers, and different other kinds of fluid/fluid separation units. This publication presents a new method for molecule sensitive, non-contact, and marker-free localized concentration mapping in vertical falling films. Using Raman spectroscopy, no label or marker is needed for the detection of the local composition in liquid mixtures. In the presented cases, the film mapping of sodium sulfate in water on a plain surface as well as an added artificial streaming disruptor with the shape of a small pyramid is scanned in three dimensions. The results show, as a prove of concept, a clear detectable spectroscopic difference between air, back plate, and sodium sulfate for every local point in all three dimensions. In conclusion, contactless Raman scanning on falling films for liquid mapping is realizable without any mechanical film interaction caused by the measuring probe. Surface gloss or optical reflections from a metallic back plate are suppressed by using only inelastic light scattering and the mathematical removal of background noise.

Visualization of Local Concentration and Viscosity Distribution during Glycerol-Water Mixing in a Y-Shape Minichannel: A Proof-of-Concept-Study.

The work presents an efficient and non-invasive method to visualize the local concentration and viscosity distribution of two miscible and non-reacting substances with a significant viscosity difference in a microchannel with a Y-shape cell. The proof-of-concept setup consists of a near-infrared (NIR) camera and cost-effective dome lighting with NIR light-emitting diodes (LED) covering the wavelength range of 1050 to 1650 nm. Absorption differences of glycerol and water and their mixtures with a mass fraction of glycerol from 0 to 0.95 gGlycgtotal-1 were analyzed in the NIR spectral area. The resulting measurement images were converted in a concentration profile by using absorbance calculated with Lambert-Beer law. A linear behavior between the concentration and the absorption coefficient is demonstrated. The result of local concentration in mass fraction was used to determine the local viscosity and illustrated as distribution images. By variating the fluid parameters, the influences of the highly different original viscosities in the mixing procedure were investigated and visualized.

Incremental Learning in Modelling Process Analysis Technology (PAT)-An Important Tool in the Measuring and Control Circuit on the Way to the Smart Factory.

To meet the demands of the chemical and pharmaceutical process industry for a combination of high measurement accuracy, product selectivity, and low cost of ownership, the existing measurement and evaluation methods have to be further developed. This paper demonstrates the attempt to combine future Raman photometers with promising evaluation methods. As part of the investigations presented here, a new and easy-to-use evaluation method based on a self-learning algorithm is presented. This method can be applied to various measurement methods and is carried out here using an example of a Raman spectrometer system and an alcohol-water mixture as demonstration fluid. The spectra's chosen bands can be later transformed to low priced and even more robust Raman photometers. The evaluation method gives more precise results than the evaluation through classical methods like one primarily used in the software package Unscrambler. This technique increases the accuracy of detection and proves the concept of Raman process monitoring for determining concentrations. In the example of alcohol/water, the computation time is less, and it can be applied to continuous column monitoring.

Rapid brain structure and tumour margin detection on whole frozen tissue sections by fast multiphotometric mid-infrared scanning.

Frozen section analysis is a frequently used method for examination of tissue samples, especially for tumour detection. In the majority of cases, the aim is to identify characteristic tissue morphologies or tumour margins. Depending on the type of tissue, a high number of misdiagnoses are associated with this process. In this work, a fast spectroscopic measurement device and workflow was developed that significantly improves the speed of whole frozen tissue section analyses and provides sufficient information to visualize tissue structures and tumour margins, dependent on their lipid and protein molecular vibrations. That optical and non-destructive method is based on selected wavenumbers in the mid-infrared (MIR) range. We present a measuring system that substantially outperforms a commercially available Fourier Transform Infrared (FT-IR) Imaging system, since it enables acquisition of reduced spectral information at a scan field of 1 cm2 in 3 s, with a spatial resolution of 20 µm. This allows fast visualization of segmented structure areas with little computational effort. For the first time, this multiphotometric MIR system is applied to biomedical tissue sections. We are referencing our novel MIR scanner on cryopreserved murine sagittal and coronal brain sections, especially focusing on the hippocampus, and show its usability for rapid identification of primary hepatocellular carcinoma (HCC) in mouse liver.

In-Line Analysis of Diffusion Processes in Micro Channels by Long Distance Raman Photometric Measurement Technology-A Proof of Concept Study.

This work presents a novel method for the non-invasive, in-line monitoring of mixing processes in microchannels using the Raman photometric technique. The measuring set-up distinguishes itself from other works in this field by utilizing recent state-of-the-art customized photon multiplier (CPM) detectors, bypassing the use of a spectrometer. This addresses the limiting factor of integration times by achieving measuring rates of 10 ms. The method was validated using the ternary system of toluene-water-acetone. The optical measuring system consists of two functional units: the coaxial Raman probe optimized for excitation at a laser wavelength of 532 nm and the photometric detector centered around the CPMs. The spot size of the focused laser is a defining factor of the spatial resolution of the set-up. The depth of focus is measured at approx. 85 µm with a spot size of approx. 45 µm, while still maintaining a relatively high numerical aperture of 0.42, the latter of which is also critical for coaxial detection of inelastically scattered photons. The working distance in this set-up is 20 mm. The microchannel is a T-junction mixer with a square cross section of 500 by 500 µm, a hydraulic diameter of 500 µm and 70 mm channel length. The extraction of acetone from toluene into water is tracked at an initial concentration of 25% as a function of flow rate and accordingly residence time. The investigated flow rates ranged from 0.1 mL/min to 0.006 mL/min. The residence times from the T-junction to the measuring point varies from 1.5 to 25 s. At 0.006 mL/min a constant acetone concentration of approx. 12.6% was measured, indicating that the mixing process reached the equilibrium of the system at approx. 12.5%. For prototype benchmarking, comparative measurements were carried out with a commercially available Raman spectrometer (RXN1, Kaiser Optical Systems, Ann Arbor, MI, USA). Count rates of the spectrophotometer surpassed those of the spectrometer by at least one order of magnitude at identical target concentrations and optical power output. The experimental data demonstrate the suitability and potential of the new measuring system to detect locally and time-resolved concentration profiles in moving fluids while avoiding external influence.

Feasibility Study for a Chemical Process Particle Size Characterization System for Explosive Environments Using Low Laser Power.

The industrial particle sensor market lacks simple, easy to use, low cost yet robust, safe and fast response solutions. Towards development of such a sensor, for in-line use in micro channels under continuous flow conditions, this work introduces static light scattering (SLS) determination of particle diameter using a laser with an emission power of less than 5 µW together with sensitive detectors with detection times of 1 ms. The measurements for the feasibility studies are made in an angular range between 20° and 160° in 2° increments. We focus on the range between 300 and 1000 nm, for applications in the production of paints, colors, pigments and crystallites. Due to the fast response time, reaction characteristics in microchannel designs for precipitation and crystallization processes can be studied. A novel method for particle diameter characterization is developed using the positions of maxima and minima and slope distribution. The novel algorithm to classify particle diameter is especially developed to be independent of dispersed phase concentration or concentration fluctuations like product flares or signal instability. Measurement signals are post processed and particle diameters are validated against Mie light scattering simulations. The design of a low cost instrument for industrial use is proposed.

Molecule Sensitive Optical Imaging and Monitoring Techniques-A Review of Applications in Micro-Process Engineering.

This paper provides an overview of how molecule-sensitive, spatially-resolved technologies can be applied for monitoring and measuring in microchannels. The principles of elastic light scattering, fluorescence, near-infrared, mid-infrared, and Raman imaging, as well as combination techniques, are briefly presented, and their advantages and disadvantages are explained. With optical methods, images can be acquired both scanning and simultaneously as a complete image. Scanning technologies require more acquisition time, and fast moving processes are not easily observable. On the other hand, molecular selectivity is very high, especially in Raman and mid-infrared (MIR) scanning. For near-infrared (NIR) images, the entire measuring range can be simultaneously recorded with indium gallium arsenide (InGaAs) cameras. However, in this wavelength range, water is the dominant molecule, so it is sometimes necessary to use complex learning algorithms that increase the preparation effort before the actual measurement. These technologies excite molecular vibrations in a variety of ways, making these methods suitable for specific products. Besides measurements of the fluid composition, technologies for particle detection are of additional importance. With scattered light techniques and evaluation according to the Mie theory, particles in the range of 0.2-1 mm can be detected, and fast growth processes can be observed. Local multispectral measurements can also be carried out with fiber optic-coupled systems through small probe heads of approximately 1 mm diameter.

Contrast enhancement of surface layers with fast middle-infrared scanning.

In this study, we present an efficient and innovative method to visualize absorption differences in the mid-infrared range with spatial resolution using laser technology. We focus on only two lasers with wavelengths between 3.4 µm and 3.6 µm and a spatial resolution of 20 µm and thus achieve a scanning speed up to 300 kS/s for fast image generation. In this article, we focus especially on the detection of C-H bands in this region of the absorption spectrum. Concealed structures are examined by calculating the measured structures with both wavelengths. In our results, we demonstrate exemplary measurements on 130-µm-thick polyvinyl chloride layers. In turn, these structures are suitable for further processing in rapid quantitative quality control.

Noncontact recognition of fluorescently labeled objects in deep tissue via a novel optical light beam arrangement.

To date, few optical imaging systems are available in clinical practice to perform noninvasive measurements transcutaneously. Instead, functional imaging is performed using ionizing radiation or intense magnetic fields in most cases. The applicability of fluorescence imaging (e.g., for the detection of fluorescently labeled objects, such as tumors) is limited due to the restricted tissue penetration of light and the required long exposure time. Thus, the development of highly sensitive and easily manageable instruments is necessary to broaden the utility of optical imaging. To advance these developments, an improved fluorescence imaging system was designed in this study that operates on the principle of noncontact laser-induced fluorescence and enables the detection of fluorescence from deeper tissue layers as well as real-time imaging. The high performance of the developed optical laser scanner results from the combination of specific point illumination, an intensified charge-coupled device (ICCD) detector with a novel light trap, and a filtering strategy. The suitability of the laser scanner was demonstrated in two representative applications and an in vivo evaluation. In addition, a comparison with a planar imaging system was performed. The results show that the exposure time with the developed laser scanner can be reduced to a few milliseconds during measurements with a penetration depth of up to 32 mm. Due to these short exposure times, real-time fluorescence imaging can be easily achieved. The ability to measure fluorescence from deep tissue layers enables clinically relevant applications, such as the detection of fluorescently labeled malignant tumors.

Gastrin-Releasing Peptide Receptor- and Prostate-Specific Membrane Antigen-Specific Ultrasmall Gold Nanoparticles for Characterization and Diagnosis of Prostate Carcinoma via Fluorescence Imaging.

Gold nanoparticles (AuNPs) have widely been used for 70 years in cancer treatment, but only in the last 15 years has the focus been on specific AuNPs with homogeneous size and shape for various areas in science. They constitute a perfect platform for multifunctionalization and therefore enable the enhancement of target affinity. Here we report on the development of tumor specific AuNPs as diagnostic tools intended for the detection of prostate cancer via fluorescence imaging and positron emission tomography (PET). The AuNPs were further evaluated in vitro and in vivo and exhibited favorable diagnostic properties concerning tumor cell uptake, biodistribution, clearance, and tumor retention.

Non-contact Raman spectroscopy for in-line monitoring of glucose and ethanol during yeast fermentations.

The monitoring of microbiological processes using Raman spectroscopy has gained in importance over the past few years. Commercial Raman spectroscopic equipment consists of a laser, spectrometer, and fiberoptic immersion probe in direct contact with the fermentation medium. To avoid possible sterilization problems and biofilm formation on the probe tip, a large-aperture Raman probe was developed. The design of the probe enables non-contact in-line measurements through glass vessels or inspection glasses of bioreactors and chemical reactors. The practical applicability of the probe was tested during yeast fermentations by monitoring the consumption of substrate glucose and the formation of ethanol as the product. Multiple linear regression models were applied to evaluate the Raman spectra. Reference values were determined by high-performance liquid chromatography. The relative errors of prediction for glucose and ethanol were 5 and 3%, respectively. The presented Raman probe allows simple adaption to a wide range of processes in the chemical, pharmaceutical, and biotechnological industries.

Highly Sensitive Raman Spectroscopy with Low Laser Power for Fast In-Line Reaction and Multiphase Flow Monitoring.

In process analytics, the applicability of Raman spectroscopy is restricted by high excitation intensities or the long integration times required. In this work, a novel Raman system was developed to minimize photon flux losses. It allows specific reduction of spectral resolution to enable the use of Raman spectroscopy for real-time analytics when strongly increased sensitivity is required. The performance potential of the optical setup was demonstrated in two exemplary applications: First, a fast exothermic reaction (Michael addition) was monitored with backscattering fiber optics under strongly attenuated laser power (7 mW). Second, high-speed scanning of a segmented multiphase flow (water/toluene) with submicroliter droplets was achieved by aligning the focus of a coaxial Raman probe with long focal length directly into a perfluoroalkoxy (PFA) capillary. With an acquisition rate of 333 Raman spectra per second, chemical information was obtained separately for both of the rapidly alternating phases. The experiment with reduced laser power demonstrates that the technique described in this paper is applicable in chemical production processes, especially in hazardous environments. Further potential uses can be envisioned in medical or biological applications with limited power input. The realization of high-speed measurements shows new possibilities for analysis of heterogeneous phase systems and of fast reactions or processes.

A Novel Analysis Technique for Transcutaneous Measurement of Glomerular Filtration Rate With Ultralow Dose Marker Concentrations.

OBJECTIVE: A novel high-precision approach [lifetime-decomposition measurement (LTDM)] for the assessment of the glomerular filtration rate (GFR) based on clearance measurements of exogenous filtration marker. METHODS: The time-correlated single photon counting (TCSPC) acquisition in combination with a new decomposition method allows the separation of signal and background from transcutaneous measurements of GFR. RESULTS: The performance of LTDM is compared versus the commercially available NIC-kidney patch-based system for transcutaneous GFR measurement. Measurements are performed in awake Sprague Dawley (SD) rats. Using the standard concentration required for the NIC-kidney system [7-mg/100-g body weight (b.w.) FITC-Sinistrin] as reference, the mean difference (bias) of the elimination curves GFR between LTDM and NIC-kidney was 4.8%. On the same animal and same day, the capability of LTDM to measure GFR with a FITC-Sinistrin dose reduced by a factor of 200 (35-µg/100-g b.w.) was tested as well. The mean differences (half lives with low dose using LTDM compared with those using first, the NIC-Kidney system and its standard concentration, and second, LTDM with the same concentration as for the NIC-Kidney system) were 3.4% and 4.5%, respectively. CONCLUSION: We demonstrate that with the LTDM strategy substantial reductions in marker concentrations are possible at the same level of accuracy. SIGNIFICANCE: LTDM aims to resolve the issue of the currently necessary large doses of fluorescence tracer required for transcutaneous GFR measurement. Due to substantially less influences from autofluorescence and artifacts, the proposed method outperforms other existing techniques for accurate percutaneous organ function measurement.