Confocal Raman Microspectroscopy Research Articles

Femtosecond-laser-inscribed cladding waveguides in KTiOPO4 crystal for second-harmonic generation and Y-branch splitters

In this study, the fabrication of straight- and Y-branch-cladding waveguides in KTiOPO4 crystals using femtosecond laser-direct writing is described. The second-harmonic generation (SHG) of green light through the cladding waveguides was realized using a pulsed-wave pump at 1,064 nm. The guiding properties of straight-cladding waveguides fabricated by varying the laser-writing conditions were investigated. The minimum insertion loss was approximately 2.0 dB at 1,064 nm. Confocal micro-Raman spectroscopy was used to investigate the lattice micro-modifications. The maximum SHG conversion efficiency of the straight-cladding waveguide reached 31.4% with a maximum output power of 79.29 mW from an input power of 252.6 mW. The performances of the Y-branch splitters with splitting angles ranging from 0.5° to 2.6° were characterized at 1,064 nm, showing excellent properties, including symmetrical output ends and approximately equal splitting ratios. An SHG conversion efficiency of 13.4% and maximum output power of 33.63 mW were achieved in the Y-branch splitter with a splitting angle of 1.0°. These findings support potential applications in constructing compact-frequency converters by using femtosecond laser-written KTiOPO4 depressed-cladding waveguides.

Optical and fluorescent properties in Er, Yb:YAG double-line waveguide by femtosecond laser direct writing

This paper reports the double-line waveguides in Er,Yb:YAG prepared by femtosecond laser (fs-laser) writing at 1030 nm, 450 fs pulse and 100 kHz repetition rate. When light propagates at 632.8 nm along transverse electric (TE) and transverse magnetic (TM) polarization, efficient single-mode guidance is demonstrated in the double-line structures. The minimum transmission loss is as low as 0.98 dB/cm in waveguide with space of 18 μm. The structural modifications in damaged tracks and strain-affected regions were characterized by confocal micro-Raman spectroscopy, and the refractive index variations were analyzed. The corresponding fluorescence emissions of Er3+ and Yb3+ are investigated in the Er,Yb:YAG double-line structure.

Dissolvable microneedles in the skin: Determination the impact of barrier disruption and dry skin on dissolution

Dissolvable microneedles (DMNs), fabricated from biocompatible materials that dissolve in both water and skin have gained popularity in dermatology. However, limited research exists on their application in compromised skin conditions. This study compares the hyaluronic acid-based DMNs penetration, formation of microchannels, dissolution, and diffusion kinetics in intact, barrier-disrupted (tape stripped), and dry (acetone-treated) porcine ear skin ex vivo. After DMNs application, comprehensive investigations including dermoscopy, stereomicroscope, skin hydration, transepidermal water loss (TEWL), optical coherence tomography (OCT), reflectance confocal laser scanning microscopy (RCLSM), confocal Raman micro-spectroscopy (CRM), two-photon tomography combined with fluorescence lifetime imaging (TPT-FLIM), histology, and scanning electron microscopy (SEM) were conducted. The 400 µm long DMNs successfully penetrated the skin to depths of ≈200 µm for dry skin and ≈200–290 µm for barrier-disrupted skin. Although DMNs fully inserted into all skin conditions, their dissolution rates were high in barrier-disrupted and low in dry skin, as observed through stereomicroscopy and TPT-FLIM. The dissolved polymer exhibited a more significant expansion in barrier-disrupted skin compared to intact skin, with the smallest increase observed in dry skin. Elevated TEWL and reduced skin hydration levels were evident in barrier-disrupted and dry skins compared to intact skin. OCT and RCLSM revealed noticeable skin indentation and pronounced microchannel areas, particularly in barrier-disrupted and dry skin. Additional confirmation of DMN effects on the skin and substance dissolution was obtained through histology, SEM, and CRM techniques. This study highlights the impact of skin condition on DMN effectiveness, emphasizing the importance of considering dissolvability and dissolution rates of needle materials, primarily composed of hyaluronic acid, for optimizing DMN-based drug delivery.

Spectroscopic Characterization of the Biochemical Profile of Mung Seedlings Following Treatment by Copper Oxide Nanoparticles

The present study aims to assess the physiological and biochemical changes in the mung plants (Vigna radiata L) due to the impact of copper oxide nanoparticles using laser-induced fluorescence, ultraviolet-visible and confocal micro-Raman spectroscopy. The mung seedlings were treated with copper oxide nanoparticles with concentrations of 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6 and 1.8 mM. The growth parameters of the mung seedlings show considerable decreases at higher concentrations of nanoparticles. The laser-induced fluorescence measurements showed a decrease in photosynthetic activity as indicated by the increase in the intensity ratio (I685/I732) with the nanoparticle concentration. Further, the ultraviolet-visible measurements show a decrease in the plant pigments chlorophyll a, chlorophyll b and carotenoids with the concentration of nanoparticles. Except for 0.2 mM, the Raman spectral signatures obtained from the leaves of mung-treated copper oxide nanoparticles reveal that the cell wall component proteins, lignin, pectin, carbohydrates, aliphatics and carotenoids declined with the treatment of nanoparticles. The decreasing intensity of carotenoids from the Raman measurements and the decreasing chlorophyll and carotenoid content in laser-induced fluorescence and ultraviolet-visible measurements suggest that copper oxide nanoparticles are toxic to mung plants. The spectroscopic techniques along with discriminatory and data processing approaches such as principal component analysis (PCA) are demonstrated to provide noninvasive, nondestructive, sensitive, rapid and effective screening of plants under various stresses.

Multimodal imaging analysis in silver fir reveals coordination in cellulose and lignin deposition.

Despite lignin being a key component of wood, the dynamics of tracheid lignification are generally overlooked in xylogenesis studies, which hampers our understanding of environmental drivers and blurs the interpretation of isotopic and anatomical signals stored in tree rings. Here, we analyzed cell wall formation in silver fir (Abies alba Mill.) tracheids to determine if cell wall lignification lags behind secondary wall deposition. For this purpose, we applied a multimodal imaging approach combining transmitted light microscopy (TLM), confocal laser scanning microscopy (CLSM), and confocal Raman microspectroscopy (RMS) on anatomical sections of wood microcores collected in northeast France on 11 dates during the 2010 growing season. Wood autofluorescence after laser excitation at 405 nm and 488 nm associated with the RMS scattering of lignin and cellulose, respectively, which allowed identification of lignifying cells (cells showing lignified and non-lignified wall fractions at the same time) in CLSM images. The number of lignifying cells in CLSM images mirrored the number of wall-thickening birefringent cells in polarized TLM images, revealing highly synchronized kinetics for wall thickening and lignification (similar timings and durations at the cell level). CLSM images and RMS chemical maps revealed a substantial incorporation of lignin into the wall at early stages of secondary wall deposition. Our results show that most of the cellulose and lignin contained in the cell wall undergo concurrent periods of deposition. This suggests a strong synchronization between cellulose and lignin-related features in conifer tree-ring records, as they originated over highly overlapped time frames.

Analysis of Skin Neoplasms’ Raman Spectra Using the Lorentz Approximation Method: Pilot Studies

Confocal Raman microspectroscopy provides the ability to diagnose cancer by quantitatively analyzing spectral features and identifying underlying biochemical changes. The differentiation of malignant skin neoplasms (basal cell carcinoma, squamous cell carcinoma), benign skin neoplasms (papilloma) and healthy skin was carried out by obtaining Raman spectra in vitro with excitation wavelengths of 532 and 785 nm. We present a new method for analyzing the parameters of spectral bands, based on the calculation of the second derivative and Lorentz approximation of the spectra. Using this method on a small selection of skin tumors, we have demonstrated that processes in skin tumors can cause deformation of the proteins’ secondary structure, leading to degradation and shift of the corresponding bands (972, 1655 cm–1) to the lower frequency. Bands corresponding to lipids in skin neoplasms either broaden and increase or split into two peaks (bands 1061, 1127, 1297, 1439, 1745 cm–1). The disruption of lipid structure, also indicated in several bands as a shift to lower wavenumbers, is possibly due to increased cell membrane fluidity in tumors. The results of the study may be useful for the development of optical biopsy methods for early diagnosis of tumors.

Energy-dependent correlation between micro/nano-morphology and stress state of femtosecond laser-induced modification in fused silica

Due to the importance of stress control in the laser processing of brittle transparent dielectrics such as glass and crystal, investigating the evolution of laser-induced stress state and its manipulation mechanism is highly desirable for ensuring high-performance crack-free laser micro/nanostructuring of dielectrics. Herein, the internal stress state of a femtosecond laser-modified fused silica glass sample at a broad range of pulse energy from 0.34 to 3.5 μJ was evaluated using a cantilever displacement method. Two obvious transition zones in the variation of microscale displacement were identified and analyzed by polarized optical microscopy, scanning electronic microscopy, and confocal micro-Raman spectroscopy. A strong energy-dependent correlation between the evolution of the micro/nano-scale morphology and stress state in laser-induced structures has been investigated and discussed. The analyzed result is beneficial for developing high-efficiency and high-quality manufacturing of 3D large-scale and high-precision glass microstructures.

Topical Peroxisome Proliferator-Activated Receptor Agonist Induces Molecular Alterations Enhancing Barrier Function and Water-Holding Capacity of the Human Stratum Corneum In Vivo

The peroxisome proliferator-activated receptor (PPAR) ligands modulate a variety of skin functions but are rarely used in cosmetics. The aim of this double-blind, placebo-controlled, in vivo study was to determine the effect of a topically applied 0.1% PPAR ligand on the composition and physiological parameters of the stratum corneum (SC). By comparing verum and placebo groups post-treatment, we demonstrate (via lipidomic analysis of tape strips) an unstatistically significant trend toward an increase in long-chain triacylglycerols (C50–C56) and medium- and long-chain ceramides (C42–C50) at the superficial SC. By comparing treated and untreated skin using confocal Raman microspectroscopy, we found that the changes in lipid composition in the verum group led to a significant increase in the number of trans conformers and orthorhombic organisation of lipids at the exemplary SC depth. An increase in unfolded states in the secondary and tertiary keratin structures results in an increased ability to bind water. The concentrations of tightly and strongly bound water increase, while weakly bound and unbound water decrease in the entire SC, indicating a transformation of water mobility to a state of increased hydrogen bonding. Thus, the topical PPAR ligands improve the water-holding capacity and the barrier function of the SC.

Second harmonic generation of visible vortex laser based on a waveguide-grating emitter in LBO.

In this work, we propose a practical solution to visible vortex laser emission at 532 nm based on second harmonic generation (SHG) in a well-designed waveguide-grating structure. Such an integrated structure is fabricated by femtosecond laser direct writing (FsLDW) in an LBO crystal. Confocal micro-Raman spectroscopy is employed for detailed analysis of FsLDW-induced localized crystalline damage. By optical excitation at 1064 nm, the guiding properties, SHG performance, as well as vortex laser generation of the waveguide-grating hybrid structure are systematically studied. Our results indicate that FsLDW waveguide-grating emitter is a reliable design holding great promise for nonlinear vortex beam generation in integrated optics.

Quantitative Confocal Raman Microspectroscopy to Evaluate Skin Penetration of Topically Applied Substances

Quantitative Confocal Raman Microspectroscopy to Evaluate Skin Penetration of Topically Applied Substances

Differential Effects of Soy Isoflavones on the Biophysical Properties of Model Membranes.

The effects that the main soy isoflavones, genistein and daidzein, have upon the biophysical properties of a model lipid bilayer composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) or DOPC with cholesterol (4 to 1 mol ratio) have been investigated by transbilayer water permeability, differential scanning calorimetry, and confocal Raman microspectroscopy. Genistein is found to increase water permeability, decrease phase transition temperature, reduce enthalpy of transition, and induce packing disorder in the DOPC membrane with an increasing concentration. On the contrary, daidzein decreases water permeability and shows negligible impact on thermodynamic parameters and packing disorder at comparable concentrations. For a cholesterol-containing DOPC bilayer, both genistein and daidzein exhibit an overall less pronounced effect on transbilayer water permeability. Their respective differential abilities to modify the physical and structural properties of biomembranes with varying lipid compositions signify a complex and sensitive nature to isoflavone interactions, which depends on the initial state of bilayer packing and the differences in the molecular structures of these soy isoflavones, and provide insights in understanding the interactions of these molecules with cellular membranes.

Distinguishing brain tumors by Label-free confocal micro-Raman spectroscopy

BackgroundBrain tumors have serious adverse effects on public health and social economy. Accurate detection of brain tumor types is critical for effective and proactive treatment, and thus improve the survival of patients. MethodsFour types of brain tumor tissue sections were detected by Raman spectroscopy. Principal component analysis (PCA) has been used to reduce the dimensionality of the Raman spectra data. Linear discriminant analysis (LDA) and quadratic discriminant analysis (QDA) methods were utilized to discriminate different types of brain tumors. ResultsRaman spectra were collected from 40 brain tumors. Variations in intensity and shift were observed in the Raman spectra positioned at 721, 854, 1004, 1032, 1128, 1248, 1449 cm−1 for different brain tumor tissues. The PCA results indicated that glioma, pituitary adenoma, and meningioma are difficult to differentiate from each other, whereas acoustic neuroma is clearly distinguished from the other three tumors. Multivariate analysis including QDA and LDA methods showed the classification accuracy rate of the QDA model was 99.47 %, better than the rate of LDA model was 95.07 %. ConclusionsRaman spectroscopy could be used to extract valuable fingerprint-type molecular and chemical information of biological samples. The demonstrated technique has the potential to be developed to a rapid, label-free, and intelligent approach to distinguish brain tumor types with high accuracy.

Migration of plasticizers from flexible PVC: Monitoring the concentration profiles by confocal Raman microspectroscopy

This study introduces and validates an effective approach for monitoring the plasticizer amount and its concentration profile in polyvinyl chloride (PVC) formulations. The method combines gravimetric measurements with confocal Raman depth profiling analysis. Several flexible PVC (pPVC) formulations were compared in this study, including: pPVC@DEHP, using the monomeric plasticizer di(2-ethylhexyl) phthalate (DEHP); pPVC@PBA, using the polymeric plasticizer poly(butylene adipate) (PBA); and crosslinked pPVC (cPVC@DEHP), where isophorone diamine (IPDA) served as the crosslinker. Migration tests were performed according to UNI EN ISO 177 standard. Spectroscopic findings were correlated with gravimetric data, highlighting strong differences in the migration behavior among the formulations. The results demonstrate the effectiveness of the proposed method in quantifying plasticizer leaching and obtaining detailed concentration profiles at fine scale.

Functionality differences between esterified and pregelatinized esterified starches simultaneously prepared by octenyl succinic anhydride modification and its application in dough

Octenyl succinic anhydride (OSA)-modified starches have gained widespread interest, but the modification can produce two starches with different states ignored. Herein, the two types of starches, esterified starch (ES) and pregelatinized esterified starch (PES), prepared by OSA modification were separated, and their structural and functional characteristics were comprehensively explored. Results showed that compared with native starch (NS), ES and PES exhibited high water-holding capacity, solubility, and swelling power and significantly decreased pasting temperature and thermal stability. Dynamic rheological tests illustrated that OSA modification changed the rheological behavior of starches. Fourier transform infrared spectroscopy confirmed that PES with higher degree of substitution showed more obvious ester carbonyl and carboxylate groups than ES. Laser confocal micro-Raman spectroscopy revealed that the short-range molecular order of ES, especially PES, decreased after modification. X-ray diffraction indicated that OSA modification disrupted the crystalline structure of starch, and that more amylose–lipid complex was formed in PES. Scanning electron microscopy showed that OSA modification eroded starchs surface and reduced its smoothness, and significantly disrupted PES integrity. ES and PES could be developed as food additives for retrogradation inhibition of dough. These results provide new insights into OSA modification and expand its functional application in foods.

Characterization of Unplasticized Polyvinyl Chloride Windows by Confocal Raman Microspectroscopy and Chemometrics

In order to nondestructively and accurately identify unplasticized polyvinyl chloride windows, a classification method based on multi-feature algorithm selection modeling was developed. Confocal Raman microspectroscopy was used to obtain the spectral profiles of 150 samples from five brands. The differences in model recognition accuracy were compared among three preprocessing methods: Savitzky-Golay filtering, Hilbert transform, and wavelet transform. The best preprocessing method was selected, and the competitive adaptive reweighted sampling was used to extract feature wavelengths. Furthermore, a Bayesian classification model was constructed to classify and differentiate each sample. The results showed that confocal Raman microspectroscopy can reflect the differences in physicochemical information among different samples. Preprocessing can improve the model’s recognition accuracy, with wavelet transform (96%) > Hilbert transform (90%) > Savitzky-Golay filtering (76%) > untreated (72%). Wavelet transform can remove noise from spectral data without changing the peak region and its absorbance. Based on wavelet transform processing and competitive adaptive reweighted sampling (CARS) combined with Bayesian discriminant analysis, a 100% accurate classification of 150 samples was successfully achieved.

Impact of a conditioning step during the treatment of wood with melamine-formaldehyde resin on dimensional stabilisation

Abstract The dimensional stabilisation of wood using thermosetting resins relies on the resin uptake into the cell walls. This study tested if a conditioning step after the impregnation and before the final heat-curing enhances the cell wall uptake to improve dimensional stabilisation without increasing the chemical consumption. Small blocks of Scots pine sapwood were vacuum-impregnated with an aqueous melamine formaldehyde solution and conditioned at 33, 70, or 95 % RH for up to 1 week before drying and curing the blocks at 103 °C. However, the conditioning step decreased the cell wall bulking and the moisture exclusion effect compared to the immediate heat curing of the impregnated samples. Analyses of the resin-treated samples by scanning electron microscopy, IR spectroscopy and confocal Raman microspectroscopy provided evidence of wood hydrolysis and polycondensation of the resin within the cell lumen during the conditioning step. Hydrolysis and removal of wood constituents may have counterbalanced the cell wall bulking of the resin. Polycondensation of the resin in the lumen increased its molecule size, which could have hindered the cell wall diffusion of the resin.

Beadlet anemone: A novel bio-indicator of microplastic pollution in the marine environment

Extensive usage of plastic in different industries and household usage has degraded to microplastic due to environmental conditions over the last year. While several researchers conducted the determination of microplastic (MP) bioaccumulation from rivers to stormwater, except for some filter-feeding species used as a bioindicator, to achieve a holistic approach to the fate of MPs in the marine system, sea anemone was used as an indicator. Microplastics were extracted from surface seawater, sediment, and sea anemones from the same sampling area and characterized. The extracted MPs were confirmed by confocal micro-Raman spectroscopy, and the morphology of the MPs was investigated by scanning electron microscopy (SEM). The results showed that the accumulation of microplastics and the type of polymer, shape, colour, and size of these persistent pollutants varied in sediment, surface water, and sea anemone. The abundance of MPs was different, and grey-coloured MPs were detected at a relatively higher level in sea anemones, from 8.7 to 13.3%, and more minor MPs (less than 0.5 mm) at 43.4–56.8% were detected in sea anemones among the sampling stations. In terms of polymer type, there are relatively more types of polymers classified in sea anemones among the sampling stations (six different types of polymers). These results indicated that the utilization of sea anemones as a bioindicator for MPs could be a critical factor in a better understanding of their pathway in the marine ecosystem. This study proved that analysing sea anemones as an indicator could offer a reliable, fast, and time-saving approach for detecting microplastic accumulation in marine systems.

Improvement of in vitro digestibility and thermostability of debranched waxy maize starch by sequential ethanol fractionation

This study aimed to improve the in vitro digestibility and thermostability of debranched waxy maize starch (DWMS) by sequential fractionation. Waxy maize starch was debranched by pullulanase, followed by sequential precipitation through controlling the ratio of starch supernatants to ethanol at 1:0.5, 1:1, and 1:1.5 (v/v). Subsequently the structural, thermal, in vitro digestive properties of DWMS were investigated. In vitro digestion results showed that the secondary ethanol fractionation of 1:1 on the basis of the initial fractionation (1:0.5) induced a significant higher amount of slowly digestive starch (SDS, 30.0 %) and resistant starch (RS, 58.6 %) amongst all three fractions, along with the highest peak temperature (Tp, 106.4 °C) and the highest decomposition value (Td, 310.0 °C) in calorimetric (DSC) and thermogravimetry (TGA) measurements. Chain length distribution, surface morphology, and laser confocal micro-Raman spectroscopy (LCM-Raman) analyses revealed that medium (degree of polymerization, DP 13– 36) and long chains (DP ≥37) respectively constituting 72.0 % and 10.2 % of DWMS resulted in the formation of spheroidal crystallites with higher homogeneity and more ordered short-range structures. Overall, this work confirmed that ethanol fractionation is an efficient method for improving the in vitro digestibility and heat stability of waxy maize starch.

Impact of e-cigarette liquid on porcine lung tissue - ex vivo confocal Raman micro-spectroscopy study.

Ex vivo porcine lung immersed in e-liquid was investigated in-depth using confocal Raman micro-spectroscopy to assess the e-liquid influence on the lung. It was found that lung-related Raman band intensities at 1002, 1548, 1618 and 1655 cm-1 increased after first and second treatments except the surface, which was attributed to the well-known optical clearing effect due to alveoli filling with e-liquid resulting in light scattering reduction. The autofluorescence enhancement was explained by oxidative stress induced in lung during exposure to e-liquid. Moreover, e-liquid induced collagen dehydration was revealed by the I937 /I926 Raman band intensity ratio change. The effect was enhanced after the second treatment of the same lung tissue that indicates the possibility of multi-step OC treatment. We hypothesize that the nicotine-flavour free e-liquids containing glycerol and propylene glycol could potentially be used in clinical protocols as optical clearing agent for enhanced in-depth Raman-guided bronchoscopy. This article is protected by copyright. All rights reserved.

Multi-analytical investigation into the materials and techniques of paintings on Northern Wei Dynasty (398–494 CE) coffin planks excavated from Shanxi, China

This study combined various non-invasive and micro-invasive analytical techniques to investigate the production process of coffin plank paintings excavated from a Northern Wei tomb in Zhijiabao Village, Datong City, Shanxi Province. Fiber-optic reflectance spectroscopy, portable Raman spectroscopy, and portable X-ray fluorescence spectroscopy were employed for in-situ non-invasive detection of coffin panel paintings. On that basis, micro-samples in smaller amounts than that of previous studies of the same type were collected and analyzed through optical microscopy, confocal micro-Raman spectroscopy, and scanning electron microscopy-energy dispersive spectroscopy. The results showed that the studied coffin plank decoration was painted after applying a layer of yellow primer to the plank surface. Specifically, cinnabar was used as the red material, pararealgar was used as the yellow color, an indigo and orpiment mixture was used as the green substance, the blue pigment was most likely an indigo and lead white mixture, gypsum was used the white material, and carbon black was used as the black pigment. Notably, the studied coffin plank was painted with a mixture of inorganic and organic materials to present the green and blue colors. This study provided not only a scientific basis for the conservation and restoration of this artifact but also new information for the research on Northern Wei art history.