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Vibrational characterization of granulosa cells from patients affected by unilateral ovarian endometriosis: New insights from infrared and Raman microspectroscopy
摘要： Endometriosis is a chronic gynaecological disease characterised by the presence of endometrial cells in extra-uterine regions. One of the main factors impacting on the fertility of women affected by endometriosis is the poor oocyte quality. Granulosa Cells (GCs) regulate oocyte development and maintain the appropriate microenvironment for the acquisition of its competence; hence, the dysregulation of these functions in GCs can lead to severe cellular damages also in oocytes. In this study, luteinized GCs samples were separately collected from both ovaries of women affected by Unilateral Ovarian Endometriosis and analyzed by infrared and Raman microspectroscopy. The spectral data were compared with those of GCs from women with diagnosis of tubal, idiopathic or male infertility (taken as control group). The coupling of these two spectroscopic techniques sheds new light on the alteration induced by this pathology on GCs metabolism and biochemical composition. In fact, the study revealed similar biochemical modifications in GCs from both ovaries of women affected by unilateral ovarian endometriosis, such as the alteration of the protein pattern, the induction of oxidative stress mechanisms, and the deregulation of lipid and carbohydrate metabolisms. These evidences suggest that unilateral endometriosis impairs the overall ovarian functions, causing alterations not only in the ovary with endometriotic lesions but also in the contralateral “healthy” one.
关键词： Unilateral Ovarian Endometriosis,Multivariate analysis,FTIR microspectroscopy,Raman microspectroscopy
更新于2025-09-23 15:23:52
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Hyperspectral holographic microscopy of bio-objects based on a modified Linnik interferometer
摘要： We present a modified Linnik microinterferometer that can register image-plane hyperspectral holograms in reflection of various microobjects in incoherent light. A voice coil was integrated into the reference arm of the interferometer to enable mirror displacement. A set of interferograms is registered in each pixel of a high-speed CMOS camera. Post processing yields spectrally resolved image-plane holograms of the object. Quantitative phase images of red blood cells are obtained for each spectral component σ = 1/λ.
关键词： microscopy,Linnik interferometer,holography,white light interferometry,Fourier transform microspectroscopy
更新于2025-09-23 15:23:52
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Carotenoids co-localize with hydroxyapatite, cholesterol, and other lipids in calcified stenotic aortic valves. Ex vivo Raman maps compared to histological patterns
摘要： Unlike its application for atherosclerotic plaque analysis, Raman microspectroscopy was sporadically used to check the sole nature of bioapatite deposits in stenotic aortic valves, neglecting the involvement of accumulated lipids/lipoproteins in the calcific process. Here, Raman microspectroscopy was employed for examination of stenotic aortic valve leaflets to add information on nature and distribution of accumulated lipids and their correlation with mineralization in the light of its potential precocious diagnostic use. Cryosections from surgically explanted stenotic aortic valves (n=4) were studied matching Raman maps against specific histological patterns. Raman maps revealed the presence of phospholipids/triglycerides and cholesterol, which showed spatial overlapping with one another and Raman-identified hydroxyapatite. Moreover, the Raman patterns correlated with those displayed by both von-Kossa-calcium- and Nile-blue-stained serial cryosections. Raman analysis also provided the first identification of carotenoids, which co-localized with the identified lipid moieties. Additional fit concerned the distribution of collagen and elastin. The good correlation of Raman maps with high-affinity staining patterns proved that Raman microspectroscopy is a reliable tool in evaluating calcification degree, alteration/displacement of extracellular matrix components, and accumulation rate of different lipid forms in calcified heart valves. In addition, the novel identification of carotenoids supports the concept that valve stenosis is an atherosclerosis-like valve lesion, consistently with their previous Raman microspectroscopical identification inside atherosclerotic plaques.
关键词： lipids,Valve calcification,Raman microspectroscopy,stenosis,carotenoids
更新于2025-09-23 15:22:29
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Determination of the microplastics emission in the effluent of a municipal waste water treatment plant using Raman microspectroscopy
摘要： Samples from the secondary clarifier effluent of a waste water treatment plant (serving 98500 inhabitants) were analyzed to determine the microplastics (MP) emission. The samples were collected using a stainless steel centrifugal pump and filtered through a 10 mm stainless steel cartridge filter. Microplastics particles (MPPs) and microplastics fibers (MPFs) were recovered by chemical and physical sample purification. To remove natural organic matter, the samples were first subjected to oxidative treatment with H2O2 and NaClO. Inorganic materials were subsequently removed by density separation in ZnCl2 (r = 1.9 g/cm3) using a centrifuge. Special centrifuge tubes were developed for this purpose. Sample analysis was performed on a Si filter by Raman micro-spectroscopy. Particles with a diameter (dp) ≥ 10 mm were analyzed. The results were differentiated by dry and wet weather samples. On average, 5900 MPPs m?3 were identified in the effluent on wet weather days compared to 3000 MPPs m?3 on dry weather days. Most of the MPPs detected were in the 30 mm < dp < 100 mm size range. The MPFs ranged between 100 mm and 1000 mm in length. While most of the MPFs were of PET origin, the MPPs consisted mainly of PET, PP, PE and PS.
关键词： Raman microspectroscopy,Oxidative treatment,Si filter,Centrifugation,Microplastics emission,WWTP effluent
更新于2025-09-23 15:22:29
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Spectral features of nuclear DNA in human sperm assessed by Raman Microspectroscopy: Effects of UV-irradiation and hydration
摘要： Raman Microspectroscopy represents an innovative tool for the assessment of sperm biochemical features otherwise undetectable by routine semen analysis. Previously, it was shown that induced DNA damage can be detected in smeared sperm by this technique. This novel readout may be of value for clinical settings especially if it can be transferred to living cells. Yet, starting with living sperms this study was carried-out using a variety of conditions to disclose the Raman features of sperm nuclei under different hydration conditions and UV exposure. Human sperm were immobilized and Raman spectra were obtained from individual sperm as repeated measurements. To create conditions with controlled DNA damage, sperm samples were exposed to ultraviolet light. Several media were used to evaluate their effect on Raman spectra in aqueous conditions. To substantiate differences between the experimental conditions, the spectra were analyzed by Principal Component Analysis. We observed that spectra of sperm nuclei obtained in different solutions showed a qualitatively unchanged spectral pattern showing the principal signals related to DNA. Evaluating the effect of ultraviolet light generated the finding that spectra representing DNA damage were only observed in dry conditions but not in aqueous medium. Thus, Raman microspectroscopy was successfully applied for sperm analysis in different conditions, among them in live spermatozoa in aqueous solution during the initial measurement, revealing the principle use of this technique. However, implementation of Raman spectroscopy as a technique for clinical sperm analysis and selection may be especially relevant when DNA evaluation can be established using live sperm.
关键词： hydration,DNA damage,sperm,UV-irradiation,Raman Microspectroscopy
更新于2025-09-23 15:21:21
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Spectroscopic Studies of M??ssbauer, Infrared, and Laser-Induced Luminescence for Classifying Rare-Earth Minerals Enriched in Iron-Rich Deposits
摘要： Rare-earth (RE) phosphates often appear as an accessory phase in igneous or metamorphic rocks; however, these rocks are composed of myriad chemical elements and nuclides that interfere with the qualitative or quantitative analyses of the RE phosphates over a range of concentrations in the absence of a pretreatment. In addition, the limit of each analytical methodology constrains the approach as well as the usefulness of the results in geoscience applications. Here, we report the specific mineral characterization of RE-containing ores from Yen Phu mine, Vietnam, using a range of state-of-the-art spectroscopic techniques in conjunction with microscopy: M?ssbauer spectroscopy, infrared microspectroscopy, time-resolved laser-induced fluorescence spectroscopy (TRLFS), and scanning electron microscopy with energy-dispersive X-ray spectroscopy. Because the distribution of each element in the deposit differs, such combinatorial works are necessary and could lead to more plausible answers to questions surrounding the point of origin of RE elements. The results of our M?ssbauer spectroscopic analysis indicate that the three ores sampled at different locations all contain magnetite-like, hematite-like, and iron(III) salts other than hematite. In addition, we confirmed the presence of phosphate around the grain boundary in the magnetite-like mineral phase by infrared microspectroscopic analysis. The present analytical findings of trace amounts of europium(III) using TRLFS suggest that the europium ions generate identical luminescence spectra despite being embedded in three different matrices of iron minerals. This demonstration highlights the benefits of combinatorial spectroscopic analyses to gain insights into the effects of the environment of REs on their solid-state chemistry and shows the potential utility of TRLFS as a resource mining tool. Further applications of this approach in the analytical screening of rocks and minerals are feasible.
关键词： energy-dispersive X-ray spectroscopy,spectroscopic techniques,rare-earth phosphates,Yen Phu mine,infrared microspectroscopy,scanning electron microscopy,time-resolved laser-induced fluorescence spectroscopy,M?ssbauer spectroscopy,Vietnam
更新于2025-09-23 15:21:01
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Editorial: Advances in Label Free Tissue Imaging With Laser Scanning Microscopy Techniques
摘要： Significant efforts are being spent at the time being for transferring various laser scanning microscopy (LSM) techniques to the realm of tissue characterization, because of their potential to circumvent some of the most important disadvantages of traditional histopathology approaches based on excisional biopsy and tissue staining. Although conventional histopathology is currently regarded as a golden standard for the diagnosing pathologies that reflect in tissular modification (e.g., cancers), limitations such as long diagnosis time, invasiveness, artifacts, sampling error, time consumption, high costs, and interpretive variability make such approaches to be impractical in many scenarios, while also placing considerable pressure on the sustainability of healthcare systems around the world. The potential of LSM techniques to contribute to overcoming these aspects derives from their “non-invasive” character. They can exploit various endogenous optical signals generated by tissues upon interaction with a laser beam and are able to provide optical sections (virtual biopsies) that reflect the tissular architecture at controlled depths. Many studies reported to date showed that LSM techniques can provide label-free information of similar pathologic relevance to the information collected for characterization/confirmation purposes with traditional histopathology approaches. These techniques are thus capable of probing optical properties of tissues with deep implications for resolving important anatomical and physiological aspects which represent hallmarks for disease predisposition and progression. To date techniques such as Confocal Laser Scanning Microscopy (CLSM) [1], Fluorescence Lifetime Imaging (FLIM) [2], Two-Photon Excited Fluorescence Microscopy (TPEF) [2–6], Second Harmonic Generation Microscopy (SHG) [5, 6], Third Harmonic Generation Microscopy (THG) [4], Coherent Anti-Stokes Raman Scattering Microscopy (CARS) [3, 7], as well as other LSM variants such as the Brillouin Microscopy [8] have already been demonstrated to be powerful tools for investigating tissue morphology, functionality, and biochemical composition with high spatial and temporal resolution. In the opinion of many, these techniques, together with investigations approaches based on their combined use, will soon become the central element of the default tissue characterization frameworks for both ex vivo and in vivo assays. Furthermore, emerging LSM techniques exploiting various ingenious strategies to achieve superresolved images in a label-free manner [9–12] are also likely to be transferred soon toward applications addressing tissue imaging.
关键词： tissue imaging,two photon excitation fluorescence microscopy,Brillouin microspectroscopy,laser scanning microscopy,second harmonic generation microscopy
更新于2025-09-23 15:19:57
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Spatiotemporal dynamics of aggregation-induced emission enhancement controlled by optical manipulation
摘要： We present spatiotemporal control of aggregation-induced emission enhancement (AIEE) of a protonated tetraphenylethene derivative by optical manipulation. A single submicrometer-sized aggregate is initially confined by laser irradiation when its fluorescence is hardly detectable. The continuous irradiation of the formed aggregate leads to sudden and rapid growth, resulting in bright yellow fluorescence emission. The fluorescence intensity at the peak wavelength of 540 nm is tremendously enhanced with growth, meaning that AIEE is activated by optical manipulation. Amazingly, the switching on/off of the activation of AIEE is arbitrarily controlled by alternating the laser power. This result means that local concentration, which increases the optical manipulation tool overcomes the electrostatic repulsion between the protonated molecules, namely, optical manipulation changes the aggregate structure. The dynamics and mechanism in AIEE controlled by optical manipulation will be discussed from the viewpoint of molecular conformation and association depending on the laser power.
关键词： Aggregation-induced emission enhancement,Fluorescence microspectroscopy,tertiary ammonium-appended tetraphenylethene,Spatiotemporal control,Optical manipulation
更新于2025-09-23 15:19:57
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Postmortem Diagnosis of Fatal Hypothermia by Fourier Transform Infrared Spectroscopic Analysis of Edema Fluid in Formalin‐Fixed, Paraffin‐Embedded Lung Tissues
摘要： The goal of this study was to investigate whether pulmonary edema could become a specific diagnostic marker for fatal hypothermia using Fourier transform infrared (FTIR) spectroscopy in combination with chemometrics. The spectral profile analysis indicated that hypothermia fatalities associated with pulmonary edema fluid contained more b-sheet protein conformational structures than the control causes of death, which included sudden cardiac death, brain injury, cerebrovascular disease, mechanical asphyxiation, intoxication, and drowning. Subsequently, the results of principal component analysis (PCA) further revealed that the content of b-sheet protein conformational structures in the pulmonary edema fluid was the main discriminatory marker between fatal hypothermia and the other causes of death. Ultimately, a robust postmortem diagnostic model for fatal hypothermia using a partial least-squares discriminant analysis (PLS-DA) algorithm was constructed. Pulmonary edema fluid spectra collected from eight new forensic autopsy cases that did not participate in the construction of the diagnostic model were predicted using the model. The results showed the causes of death of all these eight cases were correctly classified. In conclusion, this preliminary study demonstrates that FTIR spectroscopy in combination with chemometrics could be a promising approach for the postmortem diagnosis of fatal hypothermia.
关键词： postmortem diagnosis,fatal hypothermia,chemometrics,pulmonary edema fluid,Fourier transform infrared microspectroscopy,partial least-squares discriminant analysis
更新于2025-09-16 10:30:52
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Design principles for sensitivity optimization in plasmonic hydrogen sensors
摘要： Palladium nanoparticles have proven to be an exceptionally suitable material for the optical detection of hydrogen gas, due to the dielectric function that changes with the hydrogen concentration. The development of a reliable, low-cost, and widely applicable hydrogen detector requires a simple optical read-out mechanism and an optimization of the lowest detectable hydrogen concentration. So-called ‘perfect absorber’-type structures, consisting of a layer of plasmonic palladium nanoantennas suspended above a metallic mirror layer, are a promising approach to realizing such sensors. The absorption of hydrogen by palladium leads to a shift of the plasmon resonance and thus to a change in the far-?eld re?ectance spectrum. The spectral change can be analyzed in detail using spectroscopic measurements, while the re?ectance change at a speci?c wavelength can be detected with a simple photometric system of a photodiode and a monochromatic light source. Here, we systematically investigate the geometry of cavity-coupled palladium nanostructures as well as the optical system concept, which enables us to formulate a set of design rules for optimizing the hydrogen sensitivity. Employing these principles, we demonstrate the robust detection of hydrogen at concentrations down to 100 ppm. Our results are not limited to hydrogen sensing, but can be applied to any type of plasmonic sensor.
关键词： Hydrogen detection,Palladium,Tailored disorder,Plasmonic sensing,Microspectroscopy,Metasurface,Perfect absorber,Fourier-plane spectroscopy
更新于2025-09-16 10:30:52