Objective:Verrucous epidermal nevus(VEN),seborrheic keratosis(SK),verruca plana(VP),verruca vulgaris(VV),and nevus sebaceous(NS)are common verrucous proliferative skin diseases with similar clinical appearances,often ...Objective:Verrucous epidermal nevus(VEN),seborrheic keratosis(SK),verruca plana(VP),verruca vulgaris(VV),and nevus sebaceous(NS)are common verrucous proliferative skin diseases with similar clinical appearances,often posing diagnostic challenges.Dermoscopy and reflectance confocal microscopy(RCM)can aid in their differentiation,yet their specific features under these tools have not been systematically described.This study aims to summarize and analyze the dermoscopic and RCM features of VEN,SK,VP,VV,and NS.Methods:A total of 121 patients with histopathologically confirmed verrucous proliferative skin diseases were enrolled.Dermoscopy and RCM imaging was used to observe and analyze the microscopic features of these conditions.Results:Under dermoscopy,the 5 diseases displayed distinct characteristics:VEN typically showed gyriform structures;SK was characterized by gyriform structures,comedo-like openings,and milia-like cysts;VP and VV featured dotted vessels and frogspawn-like structures;NS presented as brownish-yellow globules.RCM revealed shared features such as hyperkeratosis and acanthosis across all 5 diseases.Specific features included gyriform structures and elongated rete ridges in VEN;pseudocysts and gyriform structures in SK;evenly distributed ring-like structures in VP;vacuolated cells and papillomatous proliferation in VV;and frogspawn-like structures in NS.Conclusion:These 5 verrucous proliferative skin conditions exhibit distinguishable features under both dermoscopy and RCM.The combination of these 2 noninvasive imaging modalities holds significant clinical value for the differential diagnosis of verrucous proliferative skin diseases.展开更多
AIM:To evaluate the effects of antiglaucoma eye drops on corneal nerves by in vivo confocal microscopy(IVCM).METHODS:This study comprised 79 patients diagnosed with glaucoma and 16 healthy control individuals.Among th...AIM:To evaluate the effects of antiglaucoma eye drops on corneal nerves by in vivo confocal microscopy(IVCM).METHODS:This study comprised 79 patients diagnosed with glaucoma and 16 healthy control individuals.Among the glaucoma patients,54 were treated with medication,while 25 remained untreated.Central corneal images were evaluated by IVCM,and then ACCMetrics was used to calculate the following parameters:corneal nerve fiber density(CNFD),branch density(CNBD),fiber length(CNFL),total branch density(CTBD),fiber area(CNFA),fiber width(CNFW),and fractal dimension(CNFrD).The correlation between IVCM parameters and drugs was evaluated using non-parametric measurements of Spearman’s rank correlation coefficient.RESULTS:The CNFD was reduced in glaucoma groups compared to healthy subjects(P<0.01).Patients using anti-glaucoma medications exhibited poorer confocal parameters compared to untreated patients.As the number of medications and usage count increased,CNFD,CNBD,CNFL,CTBD,CNFA,and CNFrD experienced a decline,while CNFW increased(all P<0.01).For the brinzolamide-therapy group,there was a significant decrease in CNFD and CNFL compared to the other monotherapy groups(P<0.001).In the absence of medication,CNFD in males was lower than that in females(P<0.05).Among patients under medication therapy,CNFD remained consistent between males and females.CONCLUSION:Antiglaucoma eye drops affect the microstructure of corneal nerves.IVCM and ACCMetrics are useful tools that could be used to evaluate the corneal nerve changes.展开更多
AIM： To evaluate the corneal cell morphology of new keratoconus patients wearing two different types of rigid gas-permeable（RGP） contact lenses for 1y.METHODS： Thirty nine eyes of 39 new keratoconus patients were ...AIM： To evaluate the corneal cell morphology of new keratoconus patients wearing two different types of rigid gas-permeable（RGP） contact lenses for 1y.METHODS： Thirty nine eyes of 39 new keratoconus patients were selected and randomly fitted with two types of RGP contact lenses.Group 1 had 21 eyes with regular rigid gas-permeable（RRGP） contact lens and rest 18 eyes were in group 2 with specially designed rigid gas-permeable（SRGP） contact lens.Corneal cell morphology was evaluated using a slit scanning confocal microscope at no-lens wear and after 1y of contact lens wearing.RESULTS： After 1y of contact lens wearing in group 1,the mean anterior and posterior stromal keratocyte density were significantly less（P=0.006 and P=0.001,respectively） compared to no-lens wear.The mean cell area of anterior and posterior stromal keratocyte were also significantly different（P=0.005 and P=0.001） from no-lens wear.The anterior and posterior stromal haze increased by 18.74% and 23.81%,respectively after 1y of contact lens wearing.Whereas in group 2,statistically significant changes were observed only in cell density ＆ area of anterior stroma（P=0.001 and P=0.001,respectively） after 1y.While,level of anterior and posterior stromal haze increased by 16.67% and 11.11% after 1y of contact lens wearing.Polymegathism and pleomorphism also increased after 1y of contact lens wearing in both the contact lens groups.CONCLUSION： Confocal microscopy observation shows the significant alterations in corneal cell morphology of keratoconic corneas wearing contact lenses especially in group 1.The type of contact lens must be carefully selected to minimize changes in corneal cell morphology.展开更多
AIM： To describe the clinical features and microstructural characteristics assessed by in vivo confocal microscopy（IVCM） in patients with ocular cicatricial pemphigoid（OCP）.· METHODS： A descriptive, uncontr...AIM： To describe the clinical features and microstructural characteristics assessed by in vivo confocal microscopy（IVCM） in patients with ocular cicatricial pemphigoid（OCP）.· METHODS： A descriptive, uncontrolled case series study. Patients diagnosed with OCP were examined by clinical history, slit-lamp biomicroscopy features and IVCM images. The results of direct immunofluorescence（DIF） biopsies and indirect immunofluorescence（IIF） were also recorded. Local and systemic immunosuppressive therapy were administered and adjusted according to response.·RESULTS： A total of 12 consecutive OCP patients（7male, 5 female; mean age 60.42 ±10.39y） were recruited.All patients exhibited bilateral progressive conjunctival scarring and recurrent chronic conjunctivitis was the most frequent clinical pattern. The mean duration of symptoms prior to diagnosis of OCP was 2.95 ±2.85y（range： 5mo to 10y）. The Foster classification varied from stage I to IV and 20 eyes（83%） were within or greater than Foster stage Ⅲ on presentation. Two of the 12patients（17%） demonstrated positive DIF; 3 of the 12（25%） patients reported positive IIF. The mean duration of the follow-up period was 20.17 ±11.88mo（range： 6 to48mo）. IVCM showed variable degrees of abnormality in the conjuctiva-cornea and conjuctival scarring was detected in all the involved eyes. Corneal stromal cell activation and dendritic cell infiltration presented asocular surface inflammation, ocular surface keratinization along with the destroyed Vogt palisades was noted in eyes with potential limbal stem cell deficiency. After treatment, remission of ocular surface inflammation was achieved in all the patients, 18 eyes（75%） remained stable, 6 eyes（25%） had recurrent conjunctivitis and cicatrization in 2 eyes（8%） was progressing.· CONCLUSION： As an autoimmune disease, OCP manifests as variable degrees of clinical and laboratory abnormalities with both local and systemic immunosuppressive treatment playing important roles in disease therapy. IVCM can be as a valuable non-invasive technique to assess ocular surface changes in a cellular level with a potential value for providing diagnostic evidence and monitoring therapeutic effects during follow-up.展开更多
Objective:Novel optical imaging modalities are under development with the goal of obtaining an“optical biopsy”to efficiently provide pathologic details.One such modality is confocal microscopy which allows in situ v...Objective:Novel optical imaging modalities are under development with the goal of obtaining an“optical biopsy”to efficiently provide pathologic details.One such modality is confocal microscopy which allows in situ visualization of cells within a layer of tissue and imaging of cellular-level structures.The goal of this study is to validate the ability of confocal microscopy to quickly and accurately differentiate between normal renal tissue and cancer.Methods:Specimens were obtained from patients who underwent robotic partial nephrectomy for renal mass.Samples of suspected normal and tumor tissue were extracted from the excised portion of the kidney and stained with acridine orange.The stained samples were imaged on a Nikon E600 C1 Confocal Microscope.The samples were then submitted for hematoxylin and eosin processing and read by an expert pathologist to provide a gold-standard diagnosis that can later be compared to the confocal images.Results:This study included 11 patients,17 tissue samples,and 118 confocal images.Of the 17 tissue samples,10 had a gold-standard diagnosis of cancer and seven were benign.Of 118 confocal images,66 had a gold-standard diagnosis of cancer and 52 were benign.Six confocal images were used as a training set to train eight observers.The observers were asked to rate the test images on a six point scale and the results were analyzed using a web based receiver operating characteristic curve calculator.The average accuracy,sensitivity,specificity,and area under the empirical receiver operating characteristic curve for this study were 91%,98%,81%,and 0.94 respectively.Conclusion:This preliminary study suggest that confocal microscopy can be used to distinguish cancer from normal tissue with high sensitivity and specificity.The observers in this study were trained quickly and on only six images.We expect even higher performance as observers become more familiar with the confocal images.展开更多
Objective To study the effect of ginsenoside Rb1 and total saponin of dipsacus asper on intracellular free calcium concentration mediated by β amyloid protein.So as to lay a foundation for developing effective Chines...Objective To study the effect of ginsenoside Rb1 and total saponin of dipsacus asper on intracellular free calcium concentration mediated by β amyloid protein.So as to lay a foundation for developing effective Chinese traditional medicine to treat Alzheimer’s disease.Methods The technique of laser scanning confocal microscopy combining primary cultured neurons was adopted to quantitatively analyze the change of [Ca 2+ ] i.Results The [Ca 2+ ] i of primary cultured hippocampal neurons was nmol·L -1 on basal levels.Control group showed obvious change of calcium vibration,[Ca 2+ ] i was elevated to nmol·L -1 .The peak of [Ca 2+ ] i of Rb1 group reached nmol·L -1 and was lower than that of control group .The tSDA group displayed distinct change of calcium vibration too,and [Ca 2+ ] i reached nmol·L -1 .There was a significant difference in [Ca 2+ ] i between control and tSDA group .Conclusion The research indicated that one of mechanisms by which Rb1 and tSDA protected the neurons was to maintain the balance of [Ca 2+ ] i.展开更多
Confocal microscopy is a method which has been increasingly used over the last decade in the study of the anterior ocular surface. The method allows testing and <em>in vivo</em> high resolution imaging of ...Confocal microscopy is a method which has been increasingly used over the last decade in the study of the anterior ocular surface. The method allows testing and <em>in vivo</em> high resolution imaging of the structures of the anterior eye segment, at a cellular level, which is close to the histological examination of tissues. The data provided by this method allow for a better understanding of both the functional and pathological processes occurring in the anterior ocular surface not only for scientific purposes but also in clinical practice. The aim of the present work is to summarize the current knowledge and applications of confocal microscopy of the anterior ocular surface.展开更多
The aim of this study was to apply the existing techniques that enable examination ofmacadamia kernels to provide a better understanding of physico-chemical properties of kernels during postharvest processing. These t...The aim of this study was to apply the existing techniques that enable examination ofmacadamia kernels to provide a better understanding of physico-chemical properties of kernels during postharvest processing. These techniques, such as X-ray tomography, could be applied for quality monitoring in the macadamia industry. The objectives of this study were to investigate the browning centre symptoms that usually occur in macadamia nuts-in-shell. The applied techniques included confocal microscopy, X-ray tomography and magnetic resonance imaging （MRI）. Five different varieties of macadamia nuts （A38, 246, 816, 842 and Daddow） were selected to include distinct characteristics, such as drop pattern and growing location. Analysis of the microstructure of kernels by confocal microscopy showed the distribution of possible brown pigment compounds as well as the distribution of lipids, carbohydrates and proteins inside macadamia cells. Physical properties data, including shell density and seed to volume ratio, were obtained by X-ray tomography. Magnetic resonance diffusion tensor imaging used in this study showed marked differences in microstructure which indicate that different varieties exhibit different microstructures expressed as fraction ofanisotropy and apparent diffusion coefficient that appear to be related to the occurrence of the brown centre. Hence, the findings of this study have potential to improve the existing postharvest techniques used in the macadamia processing industry. They will be of benefit to the industry in terms of improved quality control and cost reduction.展开更多
Live cell imaging methods provide important insights into the dynamics of cellular processes that cannot be derived easily from population-averaged datasets.In the bioenergy field,much research is focused on fermentat...Live cell imaging methods provide important insights into the dynamics of cellular processes that cannot be derived easily from population-averaged datasets.In the bioenergy field,much research is focused on fermentation of cellulosic biomass by thermophilic microbes to produce biofuels;however,little effort is dedicated to the development of imaging tools to monitor this dynamic biological process.This is,in part,due to the experimental challenges of imaging cells under both anaerobic and thermophilic conditions.Here an imaging system is described that integrates confocal microscopy,a flow cell device,and a lipophilic dye to visualize cells.Solutions to technical obstacles regarding suitable fluorescent markers,photodamage during imaging,and maintenance of environmental conditions during imaging are presented.This system was utilized to observe cellulose colonization by Clostridium thermocellum under anaerobic conditions at 60℃.This method enables live cell imaging of bacterial growth under anaerobic and thermophilic conditions and should be widely applicable to visualizing different cell types or processes in real time.展开更多
The characteristics of nonmetallic inclusions formed during steel production have a significant influence on steel performance.In this paper,studies on inclusions using confocal scanning laser microscopy(CSLM)are revi...The characteristics of nonmetallic inclusions formed during steel production have a significant influence on steel performance.In this paper,studies on inclusions using confocal scanning laser microscopy(CSLM)are reviewed and summarized,particularly the col-lision of various inclusions,dissolution of inclusions in liquid slag,and reactions between inclusions and steel.Solid inclusions exhibited a high collision tendency,whereas pure liquid inclusions exhibited minimal collisions because of the small attraction force induced by their<90°contact angle with molten steel.The collision of complex inclusions in molten steel was not included in the scope of this study and should be evaluated in future studies.Higher CaO/Al_(2)O_(3)and CaO/SiO_(2)ratios in liquid slag promoted the dissolution of Al_(2)O_(3)-based in-clusions.The formation of solid phases in the slag should be prevented to improve dissolution of inclusions.To accurately simulate the dissolution of inclusions in liquid slag,in-situ observation of the dissolution of inclusions at the steel-slag interface is necessary.Using a combination of CSLM and scanning electron microscopy-energy dispersive spectroscopy,the composition and morphological evolution of the inclusions during their modification by the dissolved elements in steel were observed and analyzed.Although the in-situ observa-tion of MnS and TiN precipitations has been widely studied,the in-situ observation of the evolution of oxide inclusions in steel during so-lidification and heating processes has rarely been reported.The effects of temperature,heating and cooling rates,and inclusion character-istics on the formation of acicular ferrites(AFs)have been widely studied.At a cooling rate of 3-5 K/s,the order of AF growth rate in-duced by different inclusions,as reported in literature,is Ti-O<Ti-Ca-Zr-Al-O<Mg-O<Ti-Zr-Al-O<Mn-Ti-Al-O<Ti-Al-O<Zr-Ti-Al-O.Further comprehensive experiments are required to investigate the quantitative relationship between the formation of AFs and inclusions.展开更多
High-temperature confocal scanning laser microscopy(HT-CSLM)is a potent methodology for investigating various phenomena in the field of metallurgy.Initially applied to the observation of solid phase transformations an...High-temperature confocal scanning laser microscopy(HT-CSLM)is a potent methodology for investigating various phenomena in the field of metallurgy.Initially applied to the observation of solid phase transformations and solidification,this method has gained traction in the field of non-metallic inclusion in steels in recent years.An overview of the experimental capabilities of HT-CSLM and the most important results of recent investigations regarding the topics of clean steel production are provided.It includes the formation of intragranular acicular ferrite(IAF)from the surface of non-metallic inclusions during the continuous cooling and heat treatment,which can be especially beneficial in the toughness of heat-affected zones of welded pieces.Furthermore,the investigation of agglomeration mechanisms of non-metallic inclusions(NMIs)in liquid steel is discussed to improve the insight into attraction forces between particles and clogging phenomena during continuous casting.Also,the dissolution of NMIs in various steelmaking slags can be observed by HT-CSLM to compare dissolution rates and mechanisms of NMI,where significant influences of temperature and chemical composition of the slag were shown.Last but not least,the experimental work regarding the interface between steel and slag is discussed,where novel techniques are currently being developed.A comprehensive summary of experimental techniques using HT-CSLM equipment to investigate different interactions of NMIs with steel and slag phases is compiled.展开更多
In scientific and industrial research, three-dimensional (3D) imaging, or depth measurement, is a critical tool that provides detailed insight into surface properties. Confocal microscopy, known for its precision in s...In scientific and industrial research, three-dimensional (3D) imaging, or depth measurement, is a critical tool that provides detailed insight into surface properties. Confocal microscopy, known for its precision in surface measurements, plays a key role in this field. However, 3D imaging based on confocal microscopy is often challenged by significant data requirements and slow measurement speeds. In this paper, we present a novel self-supervised learning algorithm called SSL Depth that overcomes these challenges. Specifically, our method exploits the feature learning capabilities of neural networks while avoiding the need for labeled data sets typically associated with supervised learning approaches. Through practical demonstrations on a commercially available confocal microscope, we find that our method not only maintains higher quality, but also significantly reduces the frequency of the z-axis sampling required for 3D imaging. This reduction results in a remarkable 16×measurement speed, with the potential for further acceleration in the future. Our methodological advance enables highly efficient and accurate 3D surface reconstructions, thereby expanding the potential applications of confocal microscopy in various scientific and industrial fields.展开更多
We present a unified electromagnetic modeling of coherence scanning interferometry,confocal microscopy,and focus variation microscopy as the most common techniques for surface topography inspection with micro-and nano...We present a unified electromagnetic modeling of coherence scanning interferometry,confocal microscopy,and focus variation microscopy as the most common techniques for surface topography inspection with micro-and nanometer resolution.The model aims at analyzing the instrument response and predicting systematic deviations.Since the main focus lies on the modeling of the microscopes,the light–surface interaction is considered,based on the Kirchhoff approximation extended to vectorial imaging theory.However,it can be replaced by rigorous methods without changing the microscope model.We demonstrate that all of the measuring instruments mentioned above can be modeled using the same theory with some adaption to the respective instrument.For validation,simulated results are confirmed by comparison with measurement results.展开更多
Background:To compare the efficacy of Ex-PRESS implantation versus trabeculectomy combined with phacoemulsification.Methods:A retrospective 12-month study on patients with coincident primary open-angle glaucoma(POAG)a...Background:To compare the efficacy of Ex-PRESS implantation versus trabeculectomy combined with phacoemulsification.Methods:A retrospective 12-month study on patients with coincident primary open-angle glaucoma(POAG)and cataract.The patients underwent combined phacoemulsification and Ex-PRESS implant(Phaco-ExPRESS,n=35)or phacotrabeculectomy(Phaco-Trab,n=35).The morphological structures of the filtering bleb were examined by slitlamp,anterior segment optical coherence tomography(AS-OCT)and in vivo confocal microscopy(IVCM).Complete success was defined as postoperative intraocular pressure(IOP)<18 mmHg without the use of anti-glaucoma medication.Qualified success was defined as postoperative IOP<18 mmHg with or without anti-glaucoma medications.The data were collected preoperatively and postoperatively at 2 weeks,1 month,3 months,6 months,and 12 months.Results:No significant difference in the variables such as age,IOP and perimetry was found between the groups of Phaco-ExPRESS and Phaco-Trab.At the one-year postoperative visit for filtering blebs,Phaco-ExPRESS increased the mean area of epithelial microcysts significantly from 0.10±0.05 to 0.20±0.09μm^(2)perμm^(2),while Phaco-Trab decreased the mean area significantly from 0.08±0.04 to 0.04±0.06μm^(2)perμm^(2).Notably,the hyperreflective dots detected by IVCM decreased by 84.9%in Phaco-ExPRESS but increased by 36.3%in Phaco-Trab.The hyperreflective dots were further identified as neutrophil-and monocyte-like cells.The number of these cells were negatively correlated with the microcysts area(r=−0.7,P<0.01)but positively associated with the grade of connective tissue(r=0.5,P<0.01).By creating different microstructural changes in the filtering blebs,Phaco-ExPRESS produced a higher complete success rate(84.9%vs.41.2%,P<0.01)and significant decrease in the number of anti-glaucoma medications(P<0.01)when compared with those in Phaco-Trab.However,the qualified success showed no significant difference between the two groups(100.0%vs.91.2%,P=0.24).Conclusions:At the one-year follow-up,Phaco-ExPRESS generated better filtering bleb with larger area of microcysts,looser connective tissues,and less inflammation than that of Phaco-Trab,providing adequate IOP control and less IOP-lowering medications.These findings indicate that Phaco-ExPRESS could be more preferred than Phaco-Trab for the treatment of patients with coincident POAG and cataract.展开更多
Objective:Extramammary Paget’s disease(EMPD)is often misdiagnosed as chronic dermatitis or eczema of the scrotum because of its clinical manifestations,leading to delayed diagnosis and treatment.Reflectance confocal ...Objective:Extramammary Paget’s disease(EMPD)is often misdiagnosed as chronic dermatitis or eczema of the scrotum because of its clinical manifestations,leading to delayed diagnosis and treatment.Reflectance confocal microscopy(RCM)and dermoscopy are widely used in clinical practice,enabling early diagnosis and treatment assessment of skin diseases,particularly which are easily misdiagnosed.This study aimed to summarize the RCM and dermoscopic features of EMPD which may provide evidence for diagnosis and evaluation of treatment effects.Methods:A retrospective analysis was conducted on skin lesions from 34 patients diagnosed with EMPD,confirmed through histopathological examination at the dermatology departments of Chengdu Second People’s Hospital and the Japan-China Friendship Hospital between January 2022 and December 2023.The analysis focused on typical changes of lesions,such as blood vessel patterns and background color under dermoscopy,as well as cellular morphology under RCM.Count data were presented as constituent ratios.Results:Among the included 34 patients,28 were male and 6 were female,and 104 skin lesions were identified.The lesions were classified into 3 types:36 red patches,41 dark red erosive patches,and 27 brown pigmented patches.Dermoscopic examination of EMPD revealed characteristic features,including milky-white globules(90.4%,94/104),irregular shiny white streaks(91.3%,95/104),dotted blood vessels(91.3%,95/104),and glomerular-like vessels(88.5%,92/104).RCM demonstrated epidermal structural disorganization(100.0%,104/104),vacuolated cells in the epidermis(94.2%,98/104),and Paget cells,either scattered or clustered(88.5%,92/104).The red macular lesions predominantly contained isolated vacuolated cells in the epidermis,the dark red erosive lesions exhibited Paget cell nests in the basal layer,and the brown pigmented lesions contained numerous dendritic cells.All the patients underwent wide surgical excision followed by RCM and dermoscopic examinations postoperatively.Conclusion:This study identified characteristic dermoscopic and RCM features in patients with EMPD,providing a foundation for noninvasive auxiliary diagnosis and guiding postoperative follow-up.Additionally,RCM characteristics of different lesion morphologies can help physicians more accurately assess surgical margins,leading to more precise surgical treatment options.展开更多
Recent technological advances in miniaturization have allowed for a confocal scanning microscope to be integrated into a conventional flexible endoscope,or into trans-endoscopic probes,a technique now known as confoca...Recent technological advances in miniaturization have allowed for a confocal scanning microscope to be integrated into a conventional flexible endoscope,or into trans-endoscopic probes,a technique now known as confocal endomicroscopy or confocal laser endomicroscopy.This newly-developed technology has enabled endoscopists to collect real-time in vivo histological images or "virtual biopsies" of the gastrointestinal mucosa during endoscopy,and has stimulated significant interest in the application of this technique in clinical gastroenterology.This review aims to evaluate the current data on the technical aspects and the utility of this new technology in clinical gastroenterology and its potential impact in the future,particularly in the screening or surveillance of gastrointestinal neoplasia.展开更多
The interphase nuclei of parenchyma cells and epidermal cells of garlic ( Allium sativum L.) clove were labelled with rabbit anti_actin antibody and FITC_conjugated goat anti_rabbit IgG antibody. The authors observ...The interphase nuclei of parenchyma cells and epidermal cells of garlic ( Allium sativum L.) clove were labelled with rabbit anti_actin antibody and FITC_conjugated goat anti_rabbit IgG antibody. The authors observed results with fluorescence microscopy and confocal laser scanning microscopy. The nuclei showed prominent green_yellow fluorescence, indicating the presence of actin in the nuclei. Fluorescence examination with TRITC_phalloidin showed distinctive red fluorescence in the nuclei, indicating that F_actin is present in the nuclei. Confocal laser scanning microscopy indicated the presence of F_actin containing network structures in the nuclei, but the network structures were absent and the nuclei still showed red fluorescence when the cells were treated with cytochalasin D before fixation; the red fluorescence in the nuclei was hard to be observed when the cells were treated with unlabelled phalloidin before the cells were stained with TRITC_phalloidin. These results indicate that F_actin is in the nuclei and forms network structures in the nuclei of garlic cells.展开更多
Background：Dry eye patients suffer from all kinds of symptoms.Sometimes,the clinical signs evaluation does not disclose any obvious difference in routine examination;in vivo confocal microscopy （IVCM） is a powerful...Background：Dry eye patients suffer from all kinds of symptoms.Sometimes,the clinical signs evaluation does not disclose any obvious difference in routine examination;in vivo confocal microscopy （IVCM） is a powerful tool for ocular surface disease.This study aimed to clarify meibomian gland （MG） alterations in dry eye patients with different symptoms and to compare the findings using IVCM.Methods：A total of sixty patients were recruited,all subjected to Ocular Surface Disease Index （OSDI） and Salisbury Eye Evaluation Questionnaire （SEEQ）,and questionnaires for the assessment of dry eye symptoms before clinical sign examinations were given to the patients.Finally,IVCM was applied to observe MG＇s structure.Statistical analysis was performed using the t-test,Mann-Whitney U-test and Spearman correlation analysis.The differences were statistically significant when P 〈 0.05.Results：In the severe symptom group,OSDI and SEEQ scores were significantly higher （P 〈 0.05） compared with the mild symptoms group.All other clinical sign examinations had no statistical difference in the two groups （P 〉 0.05）.However,all the IVCM-observed data showed that patients with severe symptoms had more significant fibrosis in MG （acinar unit area 691.87 ± 182.01 μm^2 for the severe,992.17 ± 170.84 μm^2 for the mild;P 〈 0.05） and severer decrease in the size of MG acinar units than those observed in patients with mild symptoms （MG acinar unit density [MGAUD] 70.08 ± 18.78 glands/mm2,MG acinar unit longest diameter [MGALD] 51.50 ± 15.51 μm,MG acinar unit shortest diameter [MGASD] 20.30 ± 11.85 μm for the severe,MGAUD 89.53 ± 39.88 glands/mm2,MGALD 81.57 ± 21.14 μm,MGASD 42.37 ± 14.55 μm for the mild;P 〈 0.05）.Dry eye symptoms were negatively correlated with MG confocal microscopic parameters and positively correlated with conjunctival inflammatory cells and Langerhans cells （P 〈 0.05）.Conclusions：IVCM application provides a strong support to differentiate dry eye patients with different symptoms：meibomian gland dysfunction （MGD） plays a pivotal role in dry eye aggravation,and using IVCM to observe MG fibrosis,changes in size and density of MG as well as status of inflammation cells can help not only correctly diagnose the type and severity of dry eye,but also possibly prognosticate in routine eye examination in the occurrence of MGD.展开更多
Objective To examine the effects of melatonin on the dynamic changes in the concentration of intracellular free Ca 2+ ([Ca 2+ ]i) in single intact cultured cortical neurons isolated from fetal rats, in order...Objective To examine the effects of melatonin on the dynamic changes in the concentration of intracellular free Ca 2+ ([Ca 2+ ]i) in single intact cultured cortical neurons isolated from fetal rats, in order to explore the possible antiaging mechanisms of melatonin (MT) Methods Using the highly fluorescent Ca 2+ sensitive indicator Fluo 3/AM, cortical neurons cultured in a 35?mm Tissue Culture Dish were in incubated for 45?min at room temperature with 5?μmol/L Fluo 3/AM, resulting in proper intracellular dye concentration to provide adequate signal strength for detection and excellent Laser Scanning Confocal Microscopy (LSCM) imaging of [Ca 2+ ]i while not disturbing normal intracellular physiology The changes in fluorescent intensity were monitored by LSCM Results Bay K8644 (10 6 ?mol/L), KCl (20 ?mmol/L), sodium L glutamate (Glu, 50?μmol/L) caused a rapid increase of [Ca 2+ ]i in cortical neurons, and this increase could be significantly attenuated by 10 6 and 10 7 mol/L MT Conclusions MT could antagonize the extracellular Ca 2+ influx, reduce Ca 2+ overload, and have a protective effect on neurons This may be one of the important antiaging mechanisms of MT展开更多
AIM： To evaluate a newly developed hand-held confocal probe for in vivo microscopic imaging of the complete gastrointestinal tract in rodents. METHODS： A novel rigid confocal probe （diameter 7 mm） was designed wit...AIM： To evaluate a newly developed hand-held confocal probe for in vivo microscopic imaging of the complete gastrointestinal tract in rodents. METHODS： A novel rigid confocal probe （diameter 7 mm） was designed with optical features similar to the flexible endomicroscopy system for use in humans using a 488 nm single line laser for fluorophore excitation, Light emission was detected at 505 to 750 nm. The field of view was 475 μm × 475 μm. Optical slice thickness was 7 μm with a lateral resolution of 0.7 μm. Subsurface serial images at different depths （surface to 250 μm） were generated in real time at 1024 × 1024 pixels （0.8 frames/s） by placing the probe onto the tissue in gentle, stable contact. Tissue specimens were sampled for histopathological correlation.RESULTS： The esophagus, stomach, small and large intestine and meso, liver, pancreas and gall bladder were visualised in vivo at high resolution in n = 48 mice. Real time microscopic imaging with the confocal minimicroscopy probe was easy to achieve. The different staining protocols （fluorescein, acriflavine, FITC-labelled dextran and L. esculentum lectin） each highlighted specific aspects of the tissue, and in vivo imaging correlated excellently with conventional histology. In vivo blood flow monitoring added a functional quality to morphologic imaging.CONCLUSION： Confocal microscopy is feasible in vivo allowing the visualisation of the complete GI tract at high resolution even of subsurface tissue structures. The new confocal probe design evaluated in this study is compatible with laparoscopy and significantly expands the field of possible applications to intra-abdominal organs. It allows immediate testing of new in vivo staining and application options and therefore permits rapid transfer from animal studies to clinical use in patients.展开更多
基金supported by the Project of Health Committee of Hunan Province(D202304128868),China.
文摘Objective:Verrucous epidermal nevus(VEN),seborrheic keratosis(SK),verruca plana(VP),verruca vulgaris(VV),and nevus sebaceous(NS)are common verrucous proliferative skin diseases with similar clinical appearances,often posing diagnostic challenges.Dermoscopy and reflectance confocal microscopy(RCM)can aid in their differentiation,yet their specific features under these tools have not been systematically described.This study aims to summarize and analyze the dermoscopic and RCM features of VEN,SK,VP,VV,and NS.Methods:A total of 121 patients with histopathologically confirmed verrucous proliferative skin diseases were enrolled.Dermoscopy and RCM imaging was used to observe and analyze the microscopic features of these conditions.Results:Under dermoscopy,the 5 diseases displayed distinct characteristics:VEN typically showed gyriform structures;SK was characterized by gyriform structures,comedo-like openings,and milia-like cysts;VP and VV featured dotted vessels and frogspawn-like structures;NS presented as brownish-yellow globules.RCM revealed shared features such as hyperkeratosis and acanthosis across all 5 diseases.Specific features included gyriform structures and elongated rete ridges in VEN;pseudocysts and gyriform structures in SK;evenly distributed ring-like structures in VP;vacuolated cells and papillomatous proliferation in VV;and frogspawn-like structures in NS.Conclusion:These 5 verrucous proliferative skin conditions exhibit distinguishable features under both dermoscopy and RCM.The combination of these 2 noninvasive imaging modalities holds significant clinical value for the differential diagnosis of verrucous proliferative skin diseases.
基金Supported by the National Natural Science Foundation of China(No.82371058)the Natural Science Foundation of Shandong Province(No.ZR2020MH172)+2 种基金the Ophthalmology New Technology Incubation Fund Program(Ophthalmology Incubation Fund Phase II Project[2022]No.[005])Medicine Science and Technology Development Program of Shandong Province(No.202107020108)Qingdao Science and Technology Beneficiary Program(No.24-1-8-smjk-16-nsh).
文摘AIM:To evaluate the effects of antiglaucoma eye drops on corneal nerves by in vivo confocal microscopy(IVCM).METHODS:This study comprised 79 patients diagnosed with glaucoma and 16 healthy control individuals.Among the glaucoma patients,54 were treated with medication,while 25 remained untreated.Central corneal images were evaluated by IVCM,and then ACCMetrics was used to calculate the following parameters:corneal nerve fiber density(CNFD),branch density(CNBD),fiber length(CNFL),total branch density(CTBD),fiber area(CNFA),fiber width(CNFW),and fractal dimension(CNFrD).The correlation between IVCM parameters and drugs was evaluated using non-parametric measurements of Spearman’s rank correlation coefficient.RESULTS:The CNFD was reduced in glaucoma groups compared to healthy subjects(P<0.01).Patients using anti-glaucoma medications exhibited poorer confocal parameters compared to untreated patients.As the number of medications and usage count increased,CNFD,CNBD,CNFL,CTBD,CNFA,and CNFrD experienced a decline,while CNFW increased(all P<0.01).For the brinzolamide-therapy group,there was a significant decrease in CNFD and CNFL compared to the other monotherapy groups(P<0.001).In the absence of medication,CNFD in males was lower than that in females(P<0.05).Among patients under medication therapy,CNFD remained consistent between males and females.CONCLUSION:Antiglaucoma eye drops affect the microstructure of corneal nerves.IVCM and ACCMetrics are useful tools that could be used to evaluate the corneal nerve changes.
文摘AIM： To evaluate the corneal cell morphology of new keratoconus patients wearing two different types of rigid gas-permeable（RGP） contact lenses for 1y.METHODS： Thirty nine eyes of 39 new keratoconus patients were selected and randomly fitted with two types of RGP contact lenses.Group 1 had 21 eyes with regular rigid gas-permeable（RRGP） contact lens and rest 18 eyes were in group 2 with specially designed rigid gas-permeable（SRGP） contact lens.Corneal cell morphology was evaluated using a slit scanning confocal microscope at no-lens wear and after 1y of contact lens wearing.RESULTS： After 1y of contact lens wearing in group 1,the mean anterior and posterior stromal keratocyte density were significantly less（P=0.006 and P=0.001,respectively） compared to no-lens wear.The mean cell area of anterior and posterior stromal keratocyte were also significantly different（P=0.005 and P=0.001） from no-lens wear.The anterior and posterior stromal haze increased by 18.74% and 23.81%,respectively after 1y of contact lens wearing.Whereas in group 2,statistically significant changes were observed only in cell density ＆ area of anterior stroma（P=0.001 and P=0.001,respectively） after 1y.While,level of anterior and posterior stromal haze increased by 16.67% and 11.11% after 1y of contact lens wearing.Polymegathism and pleomorphism also increased after 1y of contact lens wearing in both the contact lens groups.CONCLUSION： Confocal microscopy observation shows the significant alterations in corneal cell morphology of keratoconic corneas wearing contact lenses especially in group 1.The type of contact lens must be carefully selected to minimize changes in corneal cell morphology.
基金Supported by the National Natural Science Foundation of China(No.81070755No.81071301)
文摘AIM： To describe the clinical features and microstructural characteristics assessed by in vivo confocal microscopy（IVCM） in patients with ocular cicatricial pemphigoid（OCP）.· METHODS： A descriptive, uncontrolled case series study. Patients diagnosed with OCP were examined by clinical history, slit-lamp biomicroscopy features and IVCM images. The results of direct immunofluorescence（DIF） biopsies and indirect immunofluorescence（IIF） were also recorded. Local and systemic immunosuppressive therapy were administered and adjusted according to response.·RESULTS： A total of 12 consecutive OCP patients（7male, 5 female; mean age 60.42 ±10.39y） were recruited.All patients exhibited bilateral progressive conjunctival scarring and recurrent chronic conjunctivitis was the most frequent clinical pattern. The mean duration of symptoms prior to diagnosis of OCP was 2.95 ±2.85y（range： 5mo to 10y）. The Foster classification varied from stage I to IV and 20 eyes（83%） were within or greater than Foster stage Ⅲ on presentation. Two of the 12patients（17%） demonstrated positive DIF; 3 of the 12（25%） patients reported positive IIF. The mean duration of the follow-up period was 20.17 ±11.88mo（range： 6 to48mo）. IVCM showed variable degrees of abnormality in the conjuctiva-cornea and conjuctival scarring was detected in all the involved eyes. Corneal stromal cell activation and dendritic cell infiltration presented asocular surface inflammation, ocular surface keratinization along with the destroyed Vogt palisades was noted in eyes with potential limbal stem cell deficiency. After treatment, remission of ocular surface inflammation was achieved in all the patients, 18 eyes（75%） remained stable, 6 eyes（25%） had recurrent conjunctivitis and cicatrization in 2 eyes（8%） was progressing.· CONCLUSION： As an autoimmune disease, OCP manifests as variable degrees of clinical and laboratory abnormalities with both local and systemic immunosuppressive treatment playing important roles in disease therapy. IVCM can be as a valuable non-invasive technique to assess ocular surface changes in a cellular level with a potential value for providing diagnostic evidence and monitoring therapeutic effects during follow-up.
基金Research reported in this publication was supported by the National Cancer Institute Cancer Center Support Grant(P30 CA023074)and used the Tissue Acquisition and Cellular/Molecular Analysis Shared Resource at the University of Arizona.
文摘Objective:Novel optical imaging modalities are under development with the goal of obtaining an“optical biopsy”to efficiently provide pathologic details.One such modality is confocal microscopy which allows in situ visualization of cells within a layer of tissue and imaging of cellular-level structures.The goal of this study is to validate the ability of confocal microscopy to quickly and accurately differentiate between normal renal tissue and cancer.Methods:Specimens were obtained from patients who underwent robotic partial nephrectomy for renal mass.Samples of suspected normal and tumor tissue were extracted from the excised portion of the kidney and stained with acridine orange.The stained samples were imaged on a Nikon E600 C1 Confocal Microscope.The samples were then submitted for hematoxylin and eosin processing and read by an expert pathologist to provide a gold-standard diagnosis that can later be compared to the confocal images.Results:This study included 11 patients,17 tissue samples,and 118 confocal images.Of the 17 tissue samples,10 had a gold-standard diagnosis of cancer and seven were benign.Of 118 confocal images,66 had a gold-standard diagnosis of cancer and 52 were benign.Six confocal images were used as a training set to train eight observers.The observers were asked to rate the test images on a six point scale and the results were analyzed using a web based receiver operating characteristic curve calculator.The average accuracy,sensitivity,specificity,and area under the empirical receiver operating characteristic curve for this study were 91%,98%,81%,and 0.94 respectively.Conclusion:This preliminary study suggest that confocal microscopy can be used to distinguish cancer from normal tissue with high sensitivity and specificity.The observers in this study were trained quickly and on only six images.We expect even higher performance as observers become more familiar with the confocal images.
文摘Objective To study the effect of ginsenoside Rb1 and total saponin of dipsacus asper on intracellular free calcium concentration mediated by β amyloid protein.So as to lay a foundation for developing effective Chinese traditional medicine to treat Alzheimer’s disease.Methods The technique of laser scanning confocal microscopy combining primary cultured neurons was adopted to quantitatively analyze the change of [Ca 2+ ] i.Results The [Ca 2+ ] i of primary cultured hippocampal neurons was nmol·L -1 on basal levels.Control group showed obvious change of calcium vibration,[Ca 2+ ] i was elevated to nmol·L -1 .The peak of [Ca 2+ ] i of Rb1 group reached nmol·L -1 and was lower than that of control group .The tSDA group displayed distinct change of calcium vibration too,and [Ca 2+ ] i reached nmol·L -1 .There was a significant difference in [Ca 2+ ] i between control and tSDA group .Conclusion The research indicated that one of mechanisms by which Rb1 and tSDA protected the neurons was to maintain the balance of [Ca 2+ ] i.
文摘Confocal microscopy is a method which has been increasingly used over the last decade in the study of the anterior ocular surface. The method allows testing and <em>in vivo</em> high resolution imaging of the structures of the anterior eye segment, at a cellular level, which is close to the histological examination of tissues. The data provided by this method allow for a better understanding of both the functional and pathological processes occurring in the anterior ocular surface not only for scientific purposes but also in clinical practice. The aim of the present work is to summarize the current knowledge and applications of confocal microscopy of the anterior ocular surface.
文摘The aim of this study was to apply the existing techniques that enable examination ofmacadamia kernels to provide a better understanding of physico-chemical properties of kernels during postharvest processing. These techniques, such as X-ray tomography, could be applied for quality monitoring in the macadamia industry. The objectives of this study were to investigate the browning centre symptoms that usually occur in macadamia nuts-in-shell. The applied techniques included confocal microscopy, X-ray tomography and magnetic resonance imaging （MRI）. Five different varieties of macadamia nuts （A38, 246, 816, 842 and Daddow） were selected to include distinct characteristics, such as drop pattern and growing location. Analysis of the microstructure of kernels by confocal microscopy showed the distribution of possible brown pigment compounds as well as the distribution of lipids, carbohydrates and proteins inside macadamia cells. Physical properties data, including shell density and seed to volume ratio, were obtained by X-ray tomography. Magnetic resonance diffusion tensor imaging used in this study showed marked differences in microstructure which indicate that different varieties exhibit different microstructures expressed as fraction ofanisotropy and apparent diffusion coefficient that appear to be related to the occurrence of the brown centre. Hence, the findings of this study have potential to improve the existing postharvest techniques used in the macadamia processing industry. They will be of benefit to the industry in terms of improved quality control and cost reduction.
基金supported by the BioEnergy Science Center (BESC),which is a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science
文摘Live cell imaging methods provide important insights into the dynamics of cellular processes that cannot be derived easily from population-averaged datasets.In the bioenergy field,much research is focused on fermentation of cellulosic biomass by thermophilic microbes to produce biofuels;however,little effort is dedicated to the development of imaging tools to monitor this dynamic biological process.This is,in part,due to the experimental challenges of imaging cells under both anaerobic and thermophilic conditions.Here an imaging system is described that integrates confocal microscopy,a flow cell device,and a lipophilic dye to visualize cells.Solutions to technical obstacles regarding suitable fluorescent markers,photodamage during imaging,and maintenance of environmental conditions during imaging are presented.This system was utilized to observe cellulose colonization by Clostridium thermocellum under anaerobic conditions at 60℃.This method enables live cell imaging of bacterial growth under anaerobic and thermophilic conditions and should be widely applicable to visualizing different cell types or processes in real time.
基金supported by the National Key R&D Program(No.2023YFB3709900)the National Nature Science Foundation of China(No.U22A20171)+2 种基金China Baowu Low Carbon Metallurgy Innovation Foundation(No.BWLCF202315)the High Steel Center(HSC)at North China University of TechnologyUniversity of Science and Technology Beijing,China.
文摘The characteristics of nonmetallic inclusions formed during steel production have a significant influence on steel performance.In this paper,studies on inclusions using confocal scanning laser microscopy(CSLM)are reviewed and summarized,particularly the col-lision of various inclusions,dissolution of inclusions in liquid slag,and reactions between inclusions and steel.Solid inclusions exhibited a high collision tendency,whereas pure liquid inclusions exhibited minimal collisions because of the small attraction force induced by their<90°contact angle with molten steel.The collision of complex inclusions in molten steel was not included in the scope of this study and should be evaluated in future studies.Higher CaO/Al_(2)O_(3)and CaO/SiO_(2)ratios in liquid slag promoted the dissolution of Al_(2)O_(3)-based in-clusions.The formation of solid phases in the slag should be prevented to improve dissolution of inclusions.To accurately simulate the dissolution of inclusions in liquid slag,in-situ observation of the dissolution of inclusions at the steel-slag interface is necessary.Using a combination of CSLM and scanning electron microscopy-energy dispersive spectroscopy,the composition and morphological evolution of the inclusions during their modification by the dissolved elements in steel were observed and analyzed.Although the in-situ observa-tion of MnS and TiN precipitations has been widely studied,the in-situ observation of the evolution of oxide inclusions in steel during so-lidification and heating processes has rarely been reported.The effects of temperature,heating and cooling rates,and inclusion character-istics on the formation of acicular ferrites(AFs)have been widely studied.At a cooling rate of 3-5 K/s,the order of AF growth rate in-duced by different inclusions,as reported in literature,is Ti-O<Ti-Ca-Zr-Al-O<Mg-O<Ti-Zr-Al-O<Mn-Ti-Al-O<Ti-Al-O<Zr-Ti-Al-O.Further comprehensive experiments are required to investigate the quantitative relationship between the formation of AFs and inclusions.
基金the Association SSF Strategic Mobility Grant(No.SM22-0039)theÅForsk Foundation(No.23-540)for supporting the research regarding inclusion engineering.
文摘High-temperature confocal scanning laser microscopy(HT-CSLM)is a potent methodology for investigating various phenomena in the field of metallurgy.Initially applied to the observation of solid phase transformations and solidification,this method has gained traction in the field of non-metallic inclusion in steels in recent years.An overview of the experimental capabilities of HT-CSLM and the most important results of recent investigations regarding the topics of clean steel production are provided.It includes the formation of intragranular acicular ferrite(IAF)from the surface of non-metallic inclusions during the continuous cooling and heat treatment,which can be especially beneficial in the toughness of heat-affected zones of welded pieces.Furthermore,the investigation of agglomeration mechanisms of non-metallic inclusions(NMIs)in liquid steel is discussed to improve the insight into attraction forces between particles and clogging phenomena during continuous casting.Also,the dissolution of NMIs in various steelmaking slags can be observed by HT-CSLM to compare dissolution rates and mechanisms of NMI,where significant influences of temperature and chemical composition of the slag were shown.Last but not least,the experimental work regarding the interface between steel and slag is discussed,where novel techniques are currently being developed.A comprehensive summary of experimental techniques using HT-CSLM equipment to investigate different interactions of NMIs with steel and slag phases is compiled.
基金supported by the Innovation Program for Quantum Science and Technology (No.2021ZD0303200)the CAS Project for Young Scientists in Basic Research (No.YSBR-049)+1 种基金the National Natural Science Foundation of China (No.62225506)the Anhui Provincial Key Research and Development Plan (No.2022b13020006)。
文摘In scientific and industrial research, three-dimensional (3D) imaging, or depth measurement, is a critical tool that provides detailed insight into surface properties. Confocal microscopy, known for its precision in surface measurements, plays a key role in this field. However, 3D imaging based on confocal microscopy is often challenged by significant data requirements and slow measurement speeds. In this paper, we present a novel self-supervised learning algorithm called SSL Depth that overcomes these challenges. Specifically, our method exploits the feature learning capabilities of neural networks while avoiding the need for labeled data sets typically associated with supervised learning approaches. Through practical demonstrations on a commercially available confocal microscope, we find that our method not only maintains higher quality, but also significantly reduces the frequency of the z-axis sampling required for 3D imaging. This reduction results in a remarkable 16×measurement speed, with the potential for further acceleration in the future. Our methodological advance enables highly efficient and accurate 3D surface reconstructions, thereby expanding the potential applications of confocal microscopy in various scientific and industrial fields.
基金support of the following research Projects (Nos.GZ:LE 992/14-3 and LE 992/18-1)by the Deutsche Forschungsgemeinschaft and the EMPIR program (project TracOptic,20IND07)co-financed by the European Union’s Horizon 2020 Research and Innovation Program.
文摘We present a unified electromagnetic modeling of coherence scanning interferometry,confocal microscopy,and focus variation microscopy as the most common techniques for surface topography inspection with micro-and nanometer resolution.The model aims at analyzing the instrument response and predicting systematic deviations.Since the main focus lies on the modeling of the microscopes,the light–surface interaction is considered,based on the Kirchhoff approximation extended to vectorial imaging theory.However,it can be replaced by rigorous methods without changing the microscope model.We demonstrate that all of the measuring instruments mentioned above can be modeled using the same theory with some adaption to the respective instrument.For validation,simulated results are confirmed by comparison with measurement results.
基金supported by the National Natural Science Foundation of Guangdong Province(Grant No.2018A030313833)supported by the National Natural Science Foundation of Guangdong Province(Grant No.2020A1515010103)supported by the National Natural Science Foundation Incubation Program of Guangdong Provincial People’s Hospital(Grant No.KY01201147 and KY01201146).
文摘Background:To compare the efficacy of Ex-PRESS implantation versus trabeculectomy combined with phacoemulsification.Methods:A retrospective 12-month study on patients with coincident primary open-angle glaucoma(POAG)and cataract.The patients underwent combined phacoemulsification and Ex-PRESS implant(Phaco-ExPRESS,n=35)or phacotrabeculectomy(Phaco-Trab,n=35).The morphological structures of the filtering bleb were examined by slitlamp,anterior segment optical coherence tomography(AS-OCT)and in vivo confocal microscopy(IVCM).Complete success was defined as postoperative intraocular pressure(IOP)<18 mmHg without the use of anti-glaucoma medication.Qualified success was defined as postoperative IOP<18 mmHg with or without anti-glaucoma medications.The data were collected preoperatively and postoperatively at 2 weeks,1 month,3 months,6 months,and 12 months.Results:No significant difference in the variables such as age,IOP and perimetry was found between the groups of Phaco-ExPRESS and Phaco-Trab.At the one-year postoperative visit for filtering blebs,Phaco-ExPRESS increased the mean area of epithelial microcysts significantly from 0.10±0.05 to 0.20±0.09μm^(2)perμm^(2),while Phaco-Trab decreased the mean area significantly from 0.08±0.04 to 0.04±0.06μm^(2)perμm^(2).Notably,the hyperreflective dots detected by IVCM decreased by 84.9%in Phaco-ExPRESS but increased by 36.3%in Phaco-Trab.The hyperreflective dots were further identified as neutrophil-and monocyte-like cells.The number of these cells were negatively correlated with the microcysts area(r=−0.7,P<0.01)but positively associated with the grade of connective tissue(r=0.5,P<0.01).By creating different microstructural changes in the filtering blebs,Phaco-ExPRESS produced a higher complete success rate(84.9%vs.41.2%,P<0.01)and significant decrease in the number of anti-glaucoma medications(P<0.01)when compared with those in Phaco-Trab.However,the qualified success showed no significant difference between the two groups(100.0%vs.91.2%,P=0.24).Conclusions:At the one-year follow-up,Phaco-ExPRESS generated better filtering bleb with larger area of microcysts,looser connective tissues,and less inflammation than that of Phaco-Trab,providing adequate IOP control and less IOP-lowering medications.These findings indicate that Phaco-ExPRESS could be more preferred than Phaco-Trab for the treatment of patients with coincident POAG and cataract.
基金supported by the National Natural Science Foundation of China Youth Project(No.82103100)the Chengdu Municipal Health Commission Research Project(No.2021201).
文摘Objective:Extramammary Paget’s disease(EMPD)is often misdiagnosed as chronic dermatitis or eczema of the scrotum because of its clinical manifestations,leading to delayed diagnosis and treatment.Reflectance confocal microscopy(RCM)and dermoscopy are widely used in clinical practice,enabling early diagnosis and treatment assessment of skin diseases,particularly which are easily misdiagnosed.This study aimed to summarize the RCM and dermoscopic features of EMPD which may provide evidence for diagnosis and evaluation of treatment effects.Methods:A retrospective analysis was conducted on skin lesions from 34 patients diagnosed with EMPD,confirmed through histopathological examination at the dermatology departments of Chengdu Second People’s Hospital and the Japan-China Friendship Hospital between January 2022 and December 2023.The analysis focused on typical changes of lesions,such as blood vessel patterns and background color under dermoscopy,as well as cellular morphology under RCM.Count data were presented as constituent ratios.Results:Among the included 34 patients,28 were male and 6 were female,and 104 skin lesions were identified.The lesions were classified into 3 types:36 red patches,41 dark red erosive patches,and 27 brown pigmented patches.Dermoscopic examination of EMPD revealed characteristic features,including milky-white globules(90.4%,94/104),irregular shiny white streaks(91.3%,95/104),dotted blood vessels(91.3%,95/104),and glomerular-like vessels(88.5%,92/104).RCM demonstrated epidermal structural disorganization(100.0%,104/104),vacuolated cells in the epidermis(94.2%,98/104),and Paget cells,either scattered or clustered(88.5%,92/104).The red macular lesions predominantly contained isolated vacuolated cells in the epidermis,the dark red erosive lesions exhibited Paget cell nests in the basal layer,and the brown pigmented lesions contained numerous dendritic cells.All the patients underwent wide surgical excision followed by RCM and dermoscopic examinations postoperatively.Conclusion:This study identified characteristic dermoscopic and RCM features in patients with EMPD,providing a foundation for noninvasive auxiliary diagnosis and guiding postoperative follow-up.Additionally,RCM characteristics of different lesion morphologies can help physicians more accurately assess surgical margins,leading to more precise surgical treatment options.
文摘Recent technological advances in miniaturization have allowed for a confocal scanning microscope to be integrated into a conventional flexible endoscope,or into trans-endoscopic probes,a technique now known as confocal endomicroscopy or confocal laser endomicroscopy.This newly-developed technology has enabled endoscopists to collect real-time in vivo histological images or "virtual biopsies" of the gastrointestinal mucosa during endoscopy,and has stimulated significant interest in the application of this technique in clinical gastroenterology.This review aims to evaluate the current data on the technical aspects and the utility of this new technology in clinical gastroenterology and its potential impact in the future,particularly in the screening or surveillance of gastrointestinal neoplasia.
文摘The interphase nuclei of parenchyma cells and epidermal cells of garlic ( Allium sativum L.) clove were labelled with rabbit anti_actin antibody and FITC_conjugated goat anti_rabbit IgG antibody. The authors observed results with fluorescence microscopy and confocal laser scanning microscopy. The nuclei showed prominent green_yellow fluorescence, indicating the presence of actin in the nuclei. Fluorescence examination with TRITC_phalloidin showed distinctive red fluorescence in the nuclei, indicating that F_actin is present in the nuclei. Confocal laser scanning microscopy indicated the presence of F_actin containing network structures in the nuclei, but the network structures were absent and the nuclei still showed red fluorescence when the cells were treated with cytochalasin D before fixation; the red fluorescence in the nuclei was hard to be observed when the cells were treated with unlabelled phalloidin before the cells were stained with TRITC_phalloidin. These results indicate that F_actin is in the nuclei and forms network structures in the nuclei of garlic cells.
文摘Background：Dry eye patients suffer from all kinds of symptoms.Sometimes,the clinical signs evaluation does not disclose any obvious difference in routine examination;in vivo confocal microscopy （IVCM） is a powerful tool for ocular surface disease.This study aimed to clarify meibomian gland （MG） alterations in dry eye patients with different symptoms and to compare the findings using IVCM.Methods：A total of sixty patients were recruited,all subjected to Ocular Surface Disease Index （OSDI） and Salisbury Eye Evaluation Questionnaire （SEEQ）,and questionnaires for the assessment of dry eye symptoms before clinical sign examinations were given to the patients.Finally,IVCM was applied to observe MG＇s structure.Statistical analysis was performed using the t-test,Mann-Whitney U-test and Spearman correlation analysis.The differences were statistically significant when P 〈 0.05.Results：In the severe symptom group,OSDI and SEEQ scores were significantly higher （P 〈 0.05） compared with the mild symptoms group.All other clinical sign examinations had no statistical difference in the two groups （P 〉 0.05）.However,all the IVCM-observed data showed that patients with severe symptoms had more significant fibrosis in MG （acinar unit area 691.87 ± 182.01 μm^2 for the severe,992.17 ± 170.84 μm^2 for the mild;P 〈 0.05） and severer decrease in the size of MG acinar units than those observed in patients with mild symptoms （MG acinar unit density [MGAUD] 70.08 ± 18.78 glands/mm2,MG acinar unit longest diameter [MGALD] 51.50 ± 15.51 μm,MG acinar unit shortest diameter [MGASD] 20.30 ± 11.85 μm for the severe,MGAUD 89.53 ± 39.88 glands/mm2,MGALD 81.57 ± 21.14 μm,MGASD 42.37 ± 14.55 μm for the mild;P 〈 0.05）.Dry eye symptoms were negatively correlated with MG confocal microscopic parameters and positively correlated with conjunctival inflammatory cells and Langerhans cells （P 〈 0.05）.Conclusions：IVCM application provides a strong support to differentiate dry eye patients with different symptoms：meibomian gland dysfunction （MGD） plays a pivotal role in dry eye aggravation,and using IVCM to observe MG fibrosis,changes in size and density of MG as well as status of inflammation cells can help not only correctly diagnose the type and severity of dry eye,but also possibly prognosticate in routine eye examination in the occurrence of MGD.
文摘Objective To examine the effects of melatonin on the dynamic changes in the concentration of intracellular free Ca 2+ ([Ca 2+ ]i) in single intact cultured cortical neurons isolated from fetal rats, in order to explore the possible antiaging mechanisms of melatonin (MT) Methods Using the highly fluorescent Ca 2+ sensitive indicator Fluo 3/AM, cortical neurons cultured in a 35?mm Tissue Culture Dish were in incubated for 45?min at room temperature with 5?μmol/L Fluo 3/AM, resulting in proper intracellular dye concentration to provide adequate signal strength for detection and excellent Laser Scanning Confocal Microscopy (LSCM) imaging of [Ca 2+ ]i while not disturbing normal intracellular physiology The changes in fluorescent intensity were monitored by LSCM Results Bay K8644 (10 6 ?mol/L), KCl (20 ?mmol/L), sodium L glutamate (Glu, 50?μmol/L) caused a rapid increase of [Ca 2+ ]i in cortical neurons, and this increase could be significantly attenuated by 10 6 and 10 7 mol/L MT Conclusions MT could antagonize the extracellular Ca 2+ influx, reduce Ca 2+ overload, and have a protective effect on neurons This may be one of the important antiaging mechanisms of MT
基金Supported by Group for Minimal-invasive Chirurgie, Johannes Gutenberg-Universitat, Mainz, Germany
文摘AIM： To evaluate a newly developed hand-held confocal probe for in vivo microscopic imaging of the complete gastrointestinal tract in rodents. METHODS： A novel rigid confocal probe （diameter 7 mm） was designed with optical features similar to the flexible endomicroscopy system for use in humans using a 488 nm single line laser for fluorophore excitation, Light emission was detected at 505 to 750 nm. The field of view was 475 μm × 475 μm. Optical slice thickness was 7 μm with a lateral resolution of 0.7 μm. Subsurface serial images at different depths （surface to 250 μm） were generated in real time at 1024 × 1024 pixels （0.8 frames/s） by placing the probe onto the tissue in gentle, stable contact. Tissue specimens were sampled for histopathological correlation.RESULTS： The esophagus, stomach, small and large intestine and meso, liver, pancreas and gall bladder were visualised in vivo at high resolution in n = 48 mice. Real time microscopic imaging with the confocal minimicroscopy probe was easy to achieve. The different staining protocols （fluorescein, acriflavine, FITC-labelled dextran and L. esculentum lectin） each highlighted specific aspects of the tissue, and in vivo imaging correlated excellently with conventional histology. In vivo blood flow monitoring added a functional quality to morphologic imaging.CONCLUSION： Confocal microscopy is feasible in vivo allowing the visualisation of the complete GI tract at high resolution even of subsurface tissue structures. The new confocal probe design evaluated in this study is compatible with laparoscopy and significantly expands the field of possible applications to intra-abdominal organs. It allows immediate testing of new in vivo staining and application options and therefore permits rapid transfer from animal studies to clinical use in patients.
