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In vivo slit scanning confocal microscopic observation in a patient with moderate and severe keratoconus: a case report
摘要： A 22-year-old Indian female was referred to Sg Buloh hospital with the diagnosis of bilateral keratoconus. On examination, slit lamp biomicroscopy and corneal topography revealed stage 3 keratoconus in the right eye and stage 2 keratoconus in the left eye. Corneal cell morphology in both eyes was evaluated using confocal microscope. In qualitative observation, almost all corneal layers in right eye except endothelium were partially or completely obscured by haze. Additionally, morphological alterations, such as elongation of keratocyte nuclei and cluster of cells, and dark bands in the anterior stroma were observed in right eye. In the left eye, the amount of haze was less, allowing better visibility of the corneal layers compared with the right eye. The dark bands were evident in the posterior stroma. Quantitative analysis showed that anterior and posterior stromal keratocyte density and endothelium cell density were relatively low in the right eye (834.0, 700.5, and 2,133 cells/mm2, respectively) compared with the left eye (934.1, 750.6, and 2,361 cells/mm2, respectively). In this case, the right eye, exhibiting stage 3 keratoconus, showed more morphological alteration, particularly in the anterior stroma compared with the left eye with stage 2 keratoconus. Increased severity of the disease can explain these differences in corneal cell morphology.
关键词： confocal microscopy,keratocyte,stromal haze,striae,cornea,keratoconus
更新于2025-09-19 17:15:36
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Reflectance confocal microscopy-guided carbon dioxide laser ablation of low-risk basal cell carcinomas: A prospective study
摘要： Background: Basal cell carcinoma (BCC) treatment modalities can be strati?ed by tumor subtype and recurrence risk. The main limitation of nonsurgical treatment modalities is the lack of histopathologic con?rmation. Re?ectance confocal microscopy (RCM) is a noninvasive imaging device that provides quasihistologic images. Objective: To evaluate the feasibility and ef?cacy of RCM-guided carbon dioxide (CO2) laser ablation of low-risk BCCs. Methods: Prospective study with biopsy specimeneproven low-risk BCCs imaged with RCM. RCM was performed on these sites before and after ablation. If residual tumor was found, a new series of laser passes were performed. The patients were then monitored for recurrence clinically and with RCM. Results: Twenty-two tumor sites in 9 patients (5 men, 4 women) were imaged and treated. Median age was 59 6 12.9 years (range, 30-74 years). Mean tumor size was 7.7 mm (range, 5-10 mm). Residual tumor was identi?ed in 5 of 22 cases (22.7%) under RCM on immediate ?rst-pass postablation sites, prompting additional laser passes. Median follow-up was 28.5 months (range, 22-32 months) with no recurrences found. Conclusions: Addition of RCM to laser ablation work?ow can detect subclinical persistent tumor after initial ablation and may serve as an aid to increase the ef?cacy of laser ablation.
关键词： laser,basal cell carcinoma,carbon dioxide laser,ablation,diagnosis,follow-up,re?ectance confocal microscopy,treatment
更新于2025-09-19 17:13:59
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Advanced multimodal laser imaging tool for urothelial carcinoma diagnosis (AMPLITUDE)
摘要： The paper presents an overview and concept of the European Union Horizon 2020 project “AMPLITUDE - Advanced Multimodal Photonics Laser Imaging Tool for Urothelial Diagnosis in Endoscopy”. The project aims at the development of a novel label-free, multi-modal imaging tool for urothelial cancer diagnosis and therapy monitoring based on ultrafast fiber laser operating in the 3rd biological window with frequency doubling and additional 785 nm CW laser for Raman spectroscopy. The tool corresponds to clinical needs for better diagnosis of the pathohistological staging of tumours and in-vivo endoscopic assessment of depth of lesion invasiveness.
关键词： 3rd biological window,multiphoton imaging,bladder cancer,confocal microscopy,short-pulsed laser
更新于2025-09-19 17:13:59
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Use of the inferior whorl to detecting age-related changes in human corneal subbasal nerve plexus with laser-scanning confocal microscopy
摘要： Purpose: To determine the effect of aging on the corneal subbasal nerve plexus (SNP) by employing a wide-field mapping technique of composite images, scanned at the location of a distinctive spiraled subbasal nerve pattern located 1 to 2 mm inferior to the corneal apex (the inferior whorl) for SNP structural quantification. Material and methods: The central corneal tactile sensitivity (CCTS) and inferior whorl length (IWL) were compared among individuals in three age groups (20–39 years, 40–59 years, and 60–79 years). Statistical analyses constituted the Kruskal-Wallis test, one-way analysis of variance (with post hoc least significant difference test), Spearman correlation coefficient, and linear regression. Results: CCTS remained stable until the age of 50 years, when it began to decrease; the mean CCTS was 58.15±2.46 mm in the group aged 20–39, 55.74±3.85 mm in the group aged 40–59, and 50.23±3.27 mm in the group aged 60–79. IWL decreased with increasing age, with a corresponding linear decline of 0.2088 mm/mm2 per year, and the mean IWL was 25.43±4.50 mm/mm2 in the group aged 20–39, 22.71±6.19 mm/mm2 in the group aged 40–59, and 18.60±4.21 mm/mm2 in the group aged 60–79. Conclusion: Our work provided a more accurate and repeatable method for corneal nerve analysis using Laser-Scanning Confocal Microscopy (LSCM). By using this technique, we confirmed that aging is associated with progressive reduction in subbasal nerve length.
关键词： aging,subbasal nerve plexus,laser-scanning confocal microscopy,inferior whorl
更新于2025-09-19 17:13:59
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Optical Fluorescence Diagnostic of Wheat Leaf Rust with Laser Scanning Confocal Microscopy
摘要： Wheat is the most important grain crop and food source worldwide. The management of diseases and early detection of pathogens is a crucial step in diagnosis programs in wheat. In the primary stage, the symptoms of rust fungus are difficult to identify with visual monitoring and other conventional techniques. In this study, we intended to investigate the early stage leaf rust in wheat crop produced through rust fungus using light fluorescence from laser scanning confocal microscopy (LSCM). The leaf rust and normal samples were analyzed with an excitation of 488 nm wavelength of Ar+ laser without any marker or photosensitizer. The small dark pores instead of stomata appears in leaf due to fungus infection and can be observed after two week of leaf tillering. These spots are orange or brown in the beginning and become black, when plants reach maturity. In recent study, the potential of non-invasive techniques for the detection of plant diseases are demonstrated for the development of a rapid and less complex early stage detection procedure that can be utilized to evaluate the infection structures during fungus infection of wheat. The newly developed rapid procedure will be helpful for early stage detection and management fungal infection before proper development during wheat interaction.
关键词： Wheat leaf rust,Optical sensors,Laser scanning confocal microscopy,Fungus infection
更新于2025-09-19 17:13:59
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The Power of Confocal Laser Scanning Microscopy in Supramolecular Chemistry: In situ Reala??time Imaging of Stimulia??Responsive Multicomponent Supramolecular Hydrogels
摘要： Multicomponent supramolecular hydrogels are promising scaffolds for applications in biosensors and controlled drug release due to their designer stimulus responsiveness. To achieve rational construction of multicomponent supramolecular hydrogel systems, their in-depth structural analysis is essential but still challenging. Confocal laser scanning microscopy (CLSM) has emerged as a powerful tool for structural analysis of multicomponent supramolecular hydrogels. CLSM imaging enables real-time observation of the hydrogels without the need of drying and/or freezing to elucidate their static and dynamic properties. Through multiple, selective fluorescent staining of materials in supramolecular hydrogels (e.g. inorganic materials and self-sorting nanofibers) can also be visualized. CLSM and the related microscopic techniques will be indispensable to investigate complex life-inspired supramolecular chemical systems.
关键词： confocal microscopy,gels,self-assembly,supramolecular chemistry,stimulus response
更新于2025-09-19 17:13:59
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The influence of MC1R dermal morphological features of photoexposed skin in women revealed by reflectance confocal microscopy and optical coherence tomography
摘要： Background: The melanocortin 1 receptor (MC1R) gene is one of the major determinants of skin pigmentation. It is a highly polymorphic gene and some of its polymorphisms, have been related to specific skin phenotypes, increased risk of skin cancers and skin photoaging. Currently, its contribution to changes in dermal features in photo-exposed skin is unknown. Objective: The main objective of this study is to evaluate the potential correlation between MC1R status and specific healthy photoexposed skin characteristics. Materials and methods: Skin facial features were estimated by evaluation with standard digital photography with automated features, reflectance confocal microscopy (RCM) and optical coherence tomography (OCT) in 100 healthy women. Skin of the forearms was used as a control. Results: The study found an association between RHC MC1R polymorphisms and dermal features in photoexposed areas being represented by increased vessel density and pixel density in OCT (p=0.025 and p=0.001, respectively) and increased coarse collagen in RCM (p=0.034), as compared to non-RHC subjects. To our knowledge this is previously unreported. Additionally, previously reported correlations between light hair and skin color and pigmented spots with MC1R RHC polymorphisms have been confirmed. Conclusions: Our results suggest the role of RHC MC1R variants in dermal variations of facial skin, as compared to non-RHC variants. To our knowledge this is previously unreported.
关键词： MC1R,melanocortin 1 receptor,reflectance confocal microscopy,skin photoaging,dynamic-optical coherence tomography
更新于2025-09-19 17:13:59
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Correlated Secondary Ion Mass Spectrometry-Laser Scanning Confocal Microscopy Imaging for Single Cell-Principles and Applications
摘要： Secondary ion mass spectrometry (SIMS) as a powerful surface analysis technique has been widely applied in semiconductor industry and geology research. Recently, with the development of instrumental technology, SIMS is attracting more and more attention in life sciences. SIMS can provide surface MS spectra, 2D/3D chemical images and depth profiling of substances simultaneously. The minimal lateral resolution of 2D SIMS imaging is 80?100 nm, and the longitudinal resolution of 3D SIMS imaging is about 1–5 nm. However, owing to lack of specific ions to render the structures of organelles, SIMS imaging for single cells still have great challenges. Optical microscopy, in particular laser scanning confocal microscopy (LSCM), has been emerged to be an indispensable technique for single cell imaging and can obtain high spatial 2D/3D imaging to visualize the structures of organelles. Thus, the combinational use of SIMS and LSCM, which takes advantages of SIMS for molecular imaging and LSCM for morphological imaging, has greatly extended the application of SIMS imaging and ensured its accuracy at single cells level, providing novel insights into better understanding of the biological events inside cells. In this review, we focus on the development and application of SIMS imaging and the correlated SIMS and LSCM imaging in the research of cell biology and drug discovery. We anticipate that the combinational use of SIMS and LSCM imaging has promising future in biomedicine and life sciences.
关键词： Cell biology,Single cell imaging,Laser scanning confocal microscopy,Correlated secondary ion mass spectrometry and laser scanning confocal microscopy imaging,Secondary ion mass spectrometry,Review
更新于2025-09-16 10:30:52
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Application of reflectance confocal microscopy to investigate the non-ablative, micro-ablative, and ablative effects of CO2 fractional laser irradiation on skin
摘要： CO2 fractional laser, as an ablative fractional laser, is commonly used in cosmetic treatment. We applied CO2 fractional laser irradiation to skin in vitro and used reflectance confocal microscopy (RCM) to image and detect the presence of any non-ablative, micro-ablative and ablative effects, in order to better understand the features of a CO2 fractional laser. In vitro irradiation of foreskin was performed using a CO2 fractional laser. Foreskin specimens were divided into 4 groups that received different amounts of irradiation energy, based on the number of irradiation passes they received: 5, 10, 15, and 20 passes, respectively. This corresponds to fluence energy of 16.3, 32.6, 48.9, 65.3 J/cm2. Immediately after irradiation, digital microscopy (DM), RCM, and histopathology were performed to observe whether the non-ablative, micro-ablative, and ablative phenomenon occurred, and the injury features of MTZs. Immediately after CO2 fractional laser irradiation, RCM and DM showed that when the numbers of passes were 5 and 10, a micro-ablative column (MAC) could not be observed or was very small. We mainly observed a thicker thermal coagulation zone (TCZ), representing non-ablative or micro-ablative effects. When the number of passes were increased to 15 and 20, the MAC was significantly enlarged and surrounded by a TCZ of medium thickness, representing ablative effects. For the first time, this study used RCM and DM to demonstrate that a CO2 fractional laser could achieve non-ablative, micro-ablative, and ablative effects on irradiated skin via different energy levels.
关键词： Microscopic treatment zone (MTZ),Non-ablative,Digital microscopy (DM),Micro-ablative,Reflectance confocal microscopy (RCM),CO2 fractional laser
更新于2025-09-12 10:27:22
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[IEEE 2019 Conference on Lasers and Electro-Optics Europe & European Quantum Electronics Conference (CLEO/Europe-EQEC) - Munich, Germany (2019.6.23-2019.6.27)] 2019 Conference on Lasers and Electro-Optics Europe & European Quantum Electronics Conference (CLEO/Europe-EQEC) - Annealing Influence on Structural and Luminescencent Properties of ZnSe:Fe
摘要： Zinc Selenide crystals doped with Iron are promising material to be used as an active media in tunable mid-IR lasers (4 ? 5 μm) [1]. The ZnSe:Fe lasing characteristics close to the theatrical maximum are obtained by optical excitation [2]. Nowadays other pumping methods are also being explored. Previously we have observed luminescence in the mid-IR spectral region with the maximum at the wavelength of 3.9 μm under electron-impact excitation. The present research is focused on the in?uence of annealing process in Zn atmosphere on defective-impurity compound (DIC) and intensity of mid-IR cathodoluminescence (CL). We studied DIC by two-photon confocal microscopy (TPCM) on Carl Zeiss LSM NLO 710 (commercially available equipment). The TPCM allows detecting edge luminescence (EL) (460-480 nm) and DIC (480 - 725 nm) as well as mapping luminescence distribution [3]. In the CL experiments, the crystals were excited by a pulsed electron gun with the accelerated electron energy of 36 kV [4]. A series of ZnSe single crystals doped with Fe by thermal diffusion from surface was subjected to the DIC and CL studies. The initial ZnSe plates were cut from boules grown from the melt by the Bridgman method with an addition of Tellurium. Iron concentrations varied from 0.01 to 14 wt.% as determined by XDR. The samples were observed by TPCM and CL before and after annealing in Zn atmosphere.
关键词： mid-IR lasers,ZnSe:Fe,two-photon confocal microscopy,cathodoluminescence,annealing
更新于2025-09-12 10:27:22