Clinical impact of confocal laser endomicroscopy in the management of gastrointestinal lesions with an uncertain diagnosis

AIM To evaluate the clinical impact of confocal laser endomicroscopy(CLE) in the diagnosis and management of patients with an uncertain diagnosis.METHODS A retrospective chart review was performed.Patients who underwent CLE between November 2013 and October 2015 and exhibited a poor correlation between endoscopic and histological findings were included.Baseline characteristics,indications,previous diagnostic studies,findings at the time of CLE,clinical management and histological results were analyzed.Interventions based on CLE findings were also analyzed.We compared the diagnostic accuracy of CLE and target biopsies of surgical specimens.RESULTS A total of 144 patients were included.Of these,51%(74/144) were female.The mean age was 51 years old.In all,41/144(28.4%) lesions were neoplastic(13 bile duct,10 gastric,8 esophageal,6 colonic,1 duodenal,1 rectal,1 ampulloma and 1 pancreatic).The sensitivity,specificity,positive predictive value,negative predictive value,and observed agreement when CLE was used to detect N-lesions were 85.37%,87.38%,72.92%,93.75% and 86.81%,respectively.Cohen's Kappa was 69.20%,thus indicating good agreement.Changes in management were observed in 54% of the cases.CONCLUSION CLE is a new diagnostic tool that has a significant clinical impact on the diagnosis and treatment of patients with uncertain diagnosis.

Neuroprotective effects of low-concentration alpha-tocopherol Confocal laser microscopy observations

BACKGROUND： Alpha-tocopherol （ α-tocopherol） can effectively relieve neuronal damage induced by oxygen-centered free radicals. However, the effective dose remains controversial. OBJECTIVE： To evaluate the protective effects of low-concentration α-tocopherol on neuronal membranes. DESIGN, TIME AND SETTING： Contrast observation and in vitro study, performed at Laboratory of Neurosurgery, Tianjin Huanhu Hospital between April and September 2006. MATERIALS： Fetal cortical neurons were derived from two 14-day pregnant SD rats, and α-tocopherol was provided by Sigma, USA. METHODS： The neurons were randomly assigned to six groups： （1） normal： neurons were cultured under normal conditions; （2） oxidative damage： oxidative free radicals was damaged using the Fenton reaction; （3） α-tocopherol： neurons were cultured in different concentrations of -tocopherol 10, 20, 40, and 80 mg/L for 2 hours, respectively. MAIN OUTCOME MEASURES： Neuronal membrane damage was observed using a confocal laser microscope, and malonaldehyde production was detected using the thiobarbituric acid method. RESULTS： At normal, biological concentrations （10 mg/L）, α-tocopherol induced no change in the damaged neurons （P 〉 0.05）. However, at a concentration of 80 mg/L, the number of damaged neurons was significantly reduced, compared with the damage group （P 〈 0.05）. Malonaldehyde levels following 80 mg/L α-tocopherol treatment were less than the oxygen free radical damage group （P 〈 0.05）, but greater than the control group （P 〈 0.01）. CONCLUSION： A concentration of 80 mg/L α-tocopherol can effectively protect the neuronal cell membrane from oxidative damage

Three-dimensional image of hepatocellular carcinoma under confocal laser scanning microscope

AIM To investigate the application of confocallaser scanning microscopy(CLSM)in tumorpathology and three-dimensional( 3-D )reconstruction by CLSM in pathologic specimensof hepatocellular carcinoma(HCC).METHODS The 30μm thick sections were cutfrom the paraffin-embedded tissues of HCC,hyperplasia and normal liver,stained with DNAfluorescent probe YOYO-1 iodide and examinedby CLSM to collect optical sections of nuclei and3-D images reconstructed.RESULTS HCC displayed chaotic arrangementof carcinoma cell nuclei,marked pleomorphism,indented and irregular nuclear surface,andirregular and coarse chromatin texture.CONCLUSION The serial optical tomograms ofCLSM can be used to create 3-D reconstruction ofcancer cell nuclei.Such 3-D impressions mightbe helpful or even essential in making anaccurate diagnosis.

Visualization of Golgia apparatus as an intracellular calcium store by laser scanning confocal microscope

Using laser scanning confocal microscopy, we have found that the in cells loaded with fluo-3/AM, highest intracellular Ca(2+) in the perinuclear region is associated with the Golgi apparatus. The spatiotemporal subcellu lar distribution of Ca(2+) in living human fibroblasts exposing to calcium-free medium in response to agonists has been investigated. PDGF, which releases Ca(2+) from intracellular stores by inositol(1, 4, 5)-trisphosphate pathway,produced a biphasic transient rise in intracellular calcium.The initial rise was resulted from a direct release of calcium from the Golgi apparatus. Calcium could be also released from and reaccumulated into the Golgi apparatus by the stimulation of thapsigargin, an inhibitor of the Ca(2+) transport ATPase of intracellular calcium store. Permeablizing the plasma membrane by 10 μM digitonin resulted in the calcium release from the Golgi apparatus and depletion of the internal calcium store. These results suggest that the Golgi apparatus plays a role in Ca(2+) regulation in signal transduction.

Confocal endomicroscopy:Is it time to move on?

Confocal laser endomicroscopy permits in-vivo microscopy evaluation during endoscopy procedures. It can be used in all the parts of the gastrointestinal tract and includes: Esophagus,stomach,small bowel,colon,biliary tract through and endoscopic retrograde cholangiopancreatography and pancreas through needles during endoscopic ultrasound procedures. Many researches demonstrated a high correlation of results between confocal laser endomicroscopy and histopathology in the diagnosis of gastrointestinal lesions; with accuracy in about 86% to 96%. Moreover,in spite that histopathology remains the gold-standard technique for final diagnosis of any diseases; a considerable number of misdiagnosis rate could be present due to many factors such as interpretation mistakes,biopsy site inaccuracy,or number of biopsies. Theoretically; with the diagnostic accuracy rates of confocal laser endomicroscopy could help in a daily practice to improve diagnosis and treatment management of the patients. However,it is still not routinely used in the clinical practice due to many factors such as cost of the procedure,lack of codification and reimbursement in some countries,absence of standard of care indications,availability,physician imageinterpretation training,medico-legal problems,and the role of the pathologist. These limitations are relative,and solutions could be found based on new researches focused to solve these barriers.

Joint Operation of Atomic Force Microscope and Advanced Laser Confocal Microscope for Observing Surface Processes in a Protein Crystal

We demonstrated the insitu observation of a moving atomic force microscope (AFM) cantilever using a laser confocal microscope combined with a differential interference microscope (LCM-DIM). The AFM cantilever scanned or indented the {110} surface of a hen egg-white lysozyme crystal in a supersaturated solution. Using a soft cantilever, we could observe the step growth with high time resolution by LCM-DIM and perform quantitative measurements of the step height by AFM simultaneously. In addition, a hard cantilever was used with LCM-DIM to observe the dynamics of crystal surface scratching and indentation. In the supersaturated solution, the small steps generated from the scratched line aggregated to macro steps, and subsequently flattened the surface.

Confocal photoluminescence characterization of silicon-vacancy color centers in 4H-SiC fabricated by a femtosecond laser

Silicon-vacancy(VSi)centers in silicon carbide(SiC)are expected to serve as solid qubits,which can be used in quantum computing and sensing.As a new controllable color center fabrication method,femtosecond(fs)laserwriting has been gradually applied in the preparation of VSi in SiC.In this study,4H-SiCwas directlywritten by an fs laser and characterized at 293 K by atomic force microscopy,confocal photoluminescence(PL),and Raman spectroscopy.PL signals of VSi were found and analyzed using 785 nm laser excitation by means of depth profiling and two-dimensional mapping.The influence of machining parameters on the VSi formation was analyzed,and the three-dimensional distribution of VSi defects in the fs laser writing of 4H-SiC was established.

Confocal laser endomicroscopy as a new diagnostic tool for poorly differentiated gastric adenocarcinoma

Gastric cancer(GC)is a multifactorial disease,where both environmental and genetic features can have an impact on its occurrence and development.GC represents one of the leading causes of cancer-related deaths worldwide.GC is most frequent in males and is believed to arise from a series of premalignant lesions.The detection of GC at an early stage is crucial because early GC,which is an invasive stomach cancer confined to the mucosal or submucosal lining,may be curable with a reported 5-year survival rate of more than 90%.Advanced GC usually has a poor prognosis despite current treatment standards.The diagnostic efficacy of conventional endoscopy(with light endoscopy)is currently limited.Confocal laser endomicroscopy is a novel imaging technique that allows real-time in vivo histological examination of mucosal surfaces during endoscopy.Confocal laser endomicroscopy may be of great importance in the surveillance of precancerous gastric lesions and in the diagnosis of GC.In this editorial we commented on the article about this topic published by Lou et al in the recent issue of the World Journal of Clinical Cases.

Dissolution behavior of Al_(2)O_(3)inclusions into CaO-MgO-SiO_(2)-Al_(2)O_(3)-TiO_(2)system ladle slags

To investigate the dissolution behaviors of Al_(2)O_(3)inclusions in CaO-5wt%MgO-SiO_(2)-30wt%Al_(2)O_(3)-TiO_(2)system ladle slags,confocal scanning laser microscopy was conducted on the slags with different TiO_(2)contents(0-10wt%),and scanning electron microscopy was performed to study the interfacial reaction between Al_(2)O_(3)and this slag system.The results disclose that the dissolution of Al_(2)O_(3)inclusions does not result in the formation of new phases at the boundary between the slag and the inclusions.In TiO_(2)-bearing and TiO_(2)-free ladle slags,there is no difference in the dissolution mechanism of Al_(2)O_(3)inclusions at steelmaking temperatures.Boundary layer diffusion is found as the controlling step of the dissolution of Al_(2)O_(3),and the diffusion coefficient is in the range of 4.18×10^(-10)to 2.18×10^(-9)m^(2)/s at 1450-1500℃.Compared with the solubility of Al_(2)O_(3)in the slags,slag viscosity and temperature play a more profound role in the dissolution of Al_(2)O_(3)inclusions.A lower viscosity and a lower melting point of the slags are beneficial for the dissolution.Suitable addition of TiO_(2)(e.g.,5wt%)in ladle slags can enhance the dissolution of Al_(2)O_(3)inclusions because of the low viscosity and melting point of the slags,while excessive addition of TiO_(2)(e.g.,10wt%)shows the opposite trend.

活体激光共聚焦显微镜观察蠕形螨感染与麦粒肿关系的临床研究

目的 采用活体激光共聚焦显微镜(IVCM)观察麦粒肿和霰粒肿患者眼部蠕形螨感染情况,为分析蠕形螨感染对麦粒肿发生的影响提供参考依据。方法 选取2022年6—12月在上海爱尔眼科医院就诊为霰粒肿的患者45例(45眼)称霰粒肿组及无霰粒肿史的麦粒肿患者45例(45眼)称麦粒肿组。每位患者采取活体激光共聚焦显微镜观察患眼睑缘蠕形螨感染情况,上下睑缘选取累及所在位置8个毛囊、8个睑板腺。以蠕形螨检出≥3只为阳性、1≤蠕形螨<3只为可疑阳性、未检出为阴性。结果 活体激光共聚焦显微镜下,麦粒肿组和霰粒肿组蠕形螨感染阳性率分别为73.30%(33眼)、44.44%(20眼),可疑阳性率为31.11%(14眼)、11.10%(5眼),阴性率为15.60%(7眼)、24.44%(11眼);麦粒肿组阳性感染率明显高于霰粒肿组,差异具有统计学意义(P=0.013)。结论 与霰粒肿患者相比,麦粒肿患者的蠕形螨感染率更高,蠕形螨感染可能亦是麦粒肿发生的致病因素之一。

Simultaneous multi-parameter observation of Harring-tonine-treating HL-60 cells with both two-photon and confocal laser scanning microscopy

Harringtonine (HT), a kind of anticancer drug isolated from Chinese herb-Cephalotaxus hainanensis Li, can induce apoptosis in promyelocytic leukemia HL-60 cells. With both two-photon laser scanning microscopy and confocal laser scanning microscopy in combination with the fluores-cent probe Hoechst 33342, tetramethyrhodamine ethyl ester (TMRE) and Fluo 3-AM, we simulta-neously observed HT-induced changes in nuclear morphology, mitochondrial membrane potential and intracellular calcium concentration ([Ca2+]i) in HL-60 cells, and developed a real-time, sensitive and invasive method for simultaneous multi-parameter observation of drug- treating living cells at the level of single cell.

Effect of melatonin on the spatial and temporal changes of [Ca^(2+) ]i in single living cells of cortical neurons by laser scanning confocal microscopy

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

Single Particle-Based Confocal Laser Scanning Microscopy for Visual Detection of Copper Ions in Confined Space

Main observation and conclusion A single particle-based confocal laser scanning microscopy was developed for the visual detection of copper ions in confined space.A fluorescence microparticle,named AuNCs/ZIF-8,was synthesized by coating gold nanoclusters(AuNCs)onto the outer surface of zeolitic imidazolate framework-8(ZIF-8).

Three-dimensional observation of the phase structure of high density polyethylene (HDPE)/poly(ethylene-co-butene) (PEB) blend by laser scanning confocal microscopy

In this paper,high density polyethylene (HDPE)/poly(ethylene-co-butene) (PEB) blend (50/50 wt%) was prepared through solution blending and then compression molding,and subsequently examined by laser scanning confocal microscopy (LSCM). The PEB used in this experiment was labeled with a small quantity of a fluorescein derivative to render fluorescence. The initial films showed uniform dye dis-tribution and no indication of phase separation within the resolution of optical microscopy. Sample films annealing at 140℃ followed by rapid cooling to room temperature showed obvious phase sepa-ration and bicontinuous structure. The present work indicates that by labeling one component with fluorescein derivative,LSCM can efficiently perform in situ depth profiling of polymer blends.

Study of bulk morphology of polymer latex films using laser confocal fluorescence microscopy

Unlabeled poly(methyl methacrylate) (PMMA) and polystyrene (PSt) latex particles as well as covalently fluorescent dye labeled PMMA latex particles (FPMMA) were prepared by emulsifier free emulsion polymerization and dispersion polymerization, respectively. The surface and bulk morphology of the polymer latex mixture (FPMMA/PMMA or FPMMA/PSt, weight ratio: 1/99) filmed by evaporation at room temperature was studied by laser confocal fluorescence microscopy (LCFM). It was concluded from the LCFM images that labeled PMMA domains obviously coagulated in PSt matrix not only on the surface but also in the inner layers of the latex film. Furthermore, PMMA was richer in the middle layers but poorer on the surface layer or the layers near to the substrate for the polymer latex films.

Laser confocal microscopy findings of Thygeson superficial punctate keratitis

Thygeson superficial punctate keratitis （TSPK） is a rare,chronic,inflammatory disorder,with adverse impact on the visual function and quality of life in patients.It was first described by Thygeson in 1950,1 as typically bilateral,corneal epithelial opacities without associated stromal involvement or corneal edema.Recurrent episodes of tearing,foreign body sensation,photophobia,and reduced vision are observed.To date,virus infection and immune factors have been considered as important risk factors for visual deterioration in the patients with TSPK,but the origin of the comeal opacities remains poorly understood.Keywords：Thygeson superficial punctate keratitis ; laser confocal microscopy

Advances in Probing Wood-Coating Interface by Microscopy: A Review

Surface coatings provide protection to wood products against weathering and other deteriorating factors, such as moisture uptake and microbial invasion. The effectiveness of coatings depends on many factors, including how well the applied coatings adhere to the wood surface. Coating adhesion to wood involves both chemical and physical interactions between the coating and wood tissues in contact, and the particular focus of this mini-review will be on the advances being made in understanding the physical aspects of the interaction by probing wood-coating interface using novel and high resolution imaging techniques, including confocal laser scanning microscopy (CLSM), SEM-backscattered electron imaging and correlative microscopy employing light, confocal and scanning electron microscopy.

苦豆子总碱二元醇脂质体制备及体外透皮特性研究

目的在制备苦豆子总碱脂质体的过程中,加入两种水相混溶渗透促进剂:乙醇和丙二醇,以获得新的、稳定的透皮给药系统,增强苦豆子总碱(SATA)的皮肤传递。方法注入法制备二元醇脂质体,并对其形态,大小,Zeta电位进行表征,包封率、囊泡稳定性和皮肤渗透特性为优化乙醇和丙二醇配比的重要考察因素,采用Franz扩散池对小鼠皮肤进行体外皮肤渗透实验,比较小鼠皮肤的累积渗透量。用激光共聚焦扫描电镜观察含有荧光素罗丹明B的苦豆子总碱二元醇脂质体在离体小鼠皮肤中的穿透深度及荧光强度。结果当乙醇∶丙二醇=7∶3(w/w)时,二元醇脂质体的稳定性最好、包封率最高(89.13±0.42)%,粒径最小(103.00±16.15)nm,透皮深度(170μm)最大和最强的荧光强度(Max FI=160 AU)。结论制备的苦豆子总碱二元醇脂质体(乙醇∶丙二醇=7∶3,w/w)是一种高效且稳定的经皮传递载体。

激光扫描共聚焦显微分析技术表征页岩亚微米级孔隙中的含油性——以准噶尔盆地芦草沟组页岩为例

亚微米级孔隙及其含油性是页岩油勘探开发所需的重要信息。为应用激光扫描共聚焦显微镜有效观测亚微米级孔隙的含油性,修改了激光扫描共聚焦显微镜主分光器和检测器的前置滤光片配置,使之能较好地进行反射光和荧光联合扫描,消除矿物所发荧光的干扰,实现了亚微米级孔隙及其含油性的精确观测。该方法在准噶尔盆地吉木萨尔凹陷芦草沟组页岩油研究中进行了应用,发现了大量的亚微米级含油孔隙;页岩油多呈不规则形状、斑块状和星点状。这套页岩的有机质成熟度Ro为0.6%~1.1%,平均约为0.9%,但轻、重质组分的分布具有显著的非均一性,不完全受成熟度控制。在有些部位,轻质组分含量及其与重质组分的比值均较高,但在成熟度相近甚至稍高的邻近部位,却存在以重质组分为主的情况。页岩层系内部的油气运移也是一个重要的控制因素。页岩中的轻质和重质组分的观测与分布规律研究,对于页岩油勘探开发十分重要,激光扫描共聚焦显微新技术可成为重要的观测手段。

激光共聚焦显微法测定面团面筋网络结构的影响因素

【目的】面团样品切片、混合染色剂浓度以及染色时间是激光共聚焦显微镜测定面团面筋网络结构的关键影响因素。本研究旨在分析这些因素对面团面筋网络结构测定结果的影响规律,进而优化出最佳的切片厚度和染色条件,为面团面筋网络结构及面制品品质高质量研究提供技术支撑。【方法】选取面筋筋力不同的两个小麦品种郑麦366(Z366)和小偃22(X22)为试验材料,磨粉制作成面团,并分别冷冻0 d和1 d;用冷冻切片机将面团分别切成10、14和20μm厚度的薄片,并配制不同浓度罗丹明B(RDB)和异硫氰酸荧光素(FITC)混合染液,分别染色1、10和20 min,分析这些因素对激光共聚焦显微法(CLSM)测定面团面筋网络结构的影响。【结果】切片厚度为10μm时,郑麦366(Z366)和小偃22(X22)面团的面筋网络结构均最清晰,且在此切片厚度下,两个品种冷冻0 d和冷冻1 d的面团面筋网络结构之间的差异均最显著(P<0.01)。染色条件研究结果表明,低浓度染色剂(0.001%+0.01%)染色时间越长,面筋蛋白网络结构呈现越清晰;高浓度染色剂(0.025%+0.25%)在染色1 min时,面筋网络结构呈现最佳,染色10 min时,面筋网络结构最差。【结论】面团切片厚度10μm,不同筋力和不同冷冻处理的面团网络结构差异最大,即面筋网络结构区分效果最明显;在此切片厚度条件下,当不考虑时间成本时,运用低浓度混合染色剂染色20 min面筋网络结构呈现效果最佳;当需要节省时间并观察淀粉颗粒细微结构时,使用高浓度染色剂染色1 min效果最好。