Clinical features and in vivo confocal microscopy assessment in 12 patients with ocular cicatricial pemphigoid

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.

Investigation of confocal microscopy for differentiation of renal cell carcinoma versus benign tissue.Can an optical biopsy be performed?

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.

Endoscopic ultrasound-guided through-the-needle microforceps biopsy and needle-based confocal laser-endomicroscopy increase detection of potentially malignant pancreatic cystic lesions:A single-center study

BACKGROUND Currently,there is insufficient data about the accuracy in the diagnosing of pancreatic cystic lesions(PCLs),especially with novel endoscopic techniques such as with direct intracystic micro-forceps biopsy(mFB)and needle-based confocal laser-endomicroscopy(nCLE).AIM To compare the accuracy of endoscopic ultrasound(EUS)and associated techniques for the detection of potentially malignant PCLs:EUS-guided fine needle aspiration(EUS-FNA),contrast-enhanced EUS(CE-EUS),EUS-guided fiberoptic probe cystoscopy(cystoscopy),mFB,and nCLE.METHODS This was a single-center,retrospective study.We identified patients who had undergone EUS,with or without additional diagnostic techniques,and had been diagnosed with PCLs.We determined agreement among malignancy after 24-mo follow-up findings with detection of potentially malignant PCLs via the EUSguided techniques and/or EUS-guided biopsy when available(EUS malignancy detection).RESULTS A total of 129 patients were included, with EUS performed alone in 47/129. In 82/129 patients,EUS procedures were performed with additional EUS-FNA (21/82), CE-EUS (20/82), cystoscopy(27/82), mFB (36/82), nCLE (44/82). Agreement between EUS malignancy detection and the 24-mo follow-up findings was higher when associated with additional diagnostic techniques thanEUS alone [62/82 (75.6%) vs 8/47 (17%);OR 4.35, 95%CI: 2.70-7.37;P < 0.001]. The highestmalignancy detection accuracy was reached when nCLE and direct intracystic mFB were bothperformed, with a sensitivity, specificity, positive predictive value, negative predictive value andobserved agreement of 100%, 89.4%, 77.8%, 100% and 92.3%, respectively (P < 0.001 comparedwith EUS-alone).CONCLUSIONThe combined use of EUS-guided mFB and nCLE improves detection of potentially malignantPCLs compared with EUS-alone, EUS-FNA, CE-EUS or cystoscopy.

EFFECT OF THE TOTAL SAPONIN OF DIPSACUS ASPER ON INTRACELLULAR FREE CALCIUM CONCENTRATION IN THE CELLULAR MODEL OF ALZHEIMER'S DISEASE-SCANNING CONFOCAL MICROSCOPY

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.

Aplication of <i>in Vivo</i>Confocal Microscopy in Ophtalmology—Overview

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 in vivo 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.

Clinical associations of corneal neuromas with ocular surface diseases

Corneal neuromas,also termed microneuromas,refer to microscopic,irregula rly-shaped enlargements of terminal subbasal nerve endings at sites of nerve damage or injury.The formation of corneal neuromas results from damage to corneal nerves,such as following corneal pathology or corneal or intraocular surge ries.Initially,denervated areas of sensory nerve fibers become invaded by sprouts of intact sensory nerve fibers,and later injured axons regenerate and new sprouts called neuromas develop.In recent years,analysis of corneal nerve abnormalities including corneal neuromas which can be identified using in vivo confocal microscopy,a non-invasive imaging technique with microscopic resolution,has been used to evaluate corneal neuropathy and ocular surface dysfunction.Corneal neuromas have been shown to be associated with clinical symptoms of discomfort and dryness of eyes,and are a promising surrogate biomarker for ocular surface diseases,such as neuropathic corneal pain,dry eye disease,diabetic corneal neuropathy,neurotrophic keratopathy,Sjogren's syndrome,bullous keratopathy,post-refra ctive surgery,and others.In this review,we have summarized the current literature on the association between these ocular surface diseases and the presentation of corneal microneuromas,as well as elaborated on their pathogenesis,visualization via in vivo confocal microscopy,and utility in monitoring treatment efficacy.As current quantitative analysis on neuromas mainly relies on manual annotation and quantification,which is user-dependent and labor-intensive,future direction includes the development of artificial intelligence software to identify and quantify these potential imaging biomarkers in a more automated and sensitive manner,allowing it to be applied in clinical settings more efficiently.Combining imaging and molecular biomarkers may also help elucidate the associations between corneal neuromas and ocular surface diseases.

Comparison of Ex‑PRESS implantation versus trabeculectomy combined with phacoemulsification in primary open‑angle glaucoma:a retrospective in vivo confocal microscopy study

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.

Long-term effects of gestational diabetes mellitus on the pancreas of female mouse offspring

BACKGROUND Prolonged fetal exposure to hyperglycemia may increase the risk of developing abnormal glucose metabolism and type-2 diabetes during childhood,adolescence,and adulthood;however,the mechanisms by which gestational diabetes mellitus(GDM)predisposes offspring to metabolic disorders remain unknown.AIM To quantify the nerve axons,macrophages,and vasculature in the pancreas from adult offspring born from mouse dams with GDM.METHODS GDM was induced by i.p.administration of streptozotocin(STZ)in ICR mouse dams.At 12 wk old,fasting blood glucose levels were determined in offspring.At 15 wk old,female offspring born from dams with and without GDM were sacrificed and pancreata were processed for immunohistochemistry.We quantified the density of sensory[calcitonin gene-related peptide(CGRP)]and tyrosine hydroxylase(TH)axons,blood vessels(endomucin),and macro-phages(CD68)in the splenic pancreas using confocal microscopy.RESULTS Offspring mice born from STZ-treated dams had similar body weight and blood glucose values compared to offspring born from vehicle-treated dams.However,the density of CGRP+and TH+axons,endomucin+blood vessels,and CD68+macrophages in the exocrine pancreas was significantly greater in offspring from mothers with GDM vs control offspring.Likewise,the microvasculature in the islets was significantly greater,but not the number of macrophages within the islets of offspring born from dams with GDM compared to control mice.CONCLUSION GDM induces neuronal,vascular,and inflammatory changes in the pancreas of adult progeny,which may partially explain the higher propensity for offspring of mothers with GDM to develop metabolic diseases.

Formation mechanisms of sub-micron pharmaceutical composite particles derived from far-and near-field Raman microscopy

Surface enhanced Raman spectroscopy(SERS) and confocal Raman microscopy are applied to investigate the structure and the molecular arrangement of sub-micron furosemide and polyvinylpyrrolidone(furosemide/PVP) particles produced by spray flash evaporation(SFE). Morphology, size and crystallinity of furosemide/PVP particles are analyzed by scanning electron microscopy(SEM) and X-ray powder diffraction(XRPD). Far-field Raman spectra and confocal far-field Raman maps of furosemide/PVP particles are interpreted based on the far-field Raman spectra of pure furosemide and PVP precursors.Confocal far-field Raman microscopy shows that furosemide/PVP particles feature an intermixture of furosemide and PVP molecules at the sub-micron scale. SERS and surface-enhanced confocal Raman microscopy(SECo RM) are performed on furosemide, PVP and furosemide/PVP composite particles sputtered with silver(40 nm). SERS and SECo RM maps reveal that furosemide/PVP particle surfaces mainly consist of PVP molecules. The combination of surface and bulk sensitive analyses reveal that furosemide/PVP sub-micron particles are formed by the agglomeration of primary furosemide nanocrystals embedded in a thin PVP matrix. Interestingly, both far-field Raman microscopy and SECo RM provide molecular information on a statistically-relevant amount of sub-micron particles in a single microscopic map;this combination is thus an effective and time-saving tool for investigating organic sub-micron composites.

THE PARALLEL CONFOCAL DETECTING SYSTEM USING OPTICAL FIBER PLATE

Objective Focusing on the problem such as slow scanning speed, complex system design and low light efficiency, a new parallel confocal 3D profile detecting method based on optical fiber technology, which realizes whole-field confocal detecting, is proposed. Methods The optical fiber plate generates an 2D point light source array, which splits one light beam into N2 subbeams and act the role of pinholes as point source and point detecting to filter the stray light and reflect light. By introducing the construction and working principle of the multi-beam 3D detecting system, the feasibility is investigated. Results Experiment result indicates that the optical fiber technology is applicable in parallel confocal detecting. Conclusion The equipment needn’t mechanical rotation. The measuring parameters that influence the detecting can easily be adapted to satisfy different requirments of measurement. Compared with the conventional confocal method, the parallel confocal detecting system using optical fiber plate is simple in the mechanism, the measuring field is larger and the speed is faster.

Advanced optical microscopy methods for in vivo imaging of sub-cellular structures in thick biological tissuesl

Optical microscopy has become an indispensable tool for visualizing sub-cellular structures andbiological processes.However,scattering in biological tissues is a major obstacle that preventshigh-resolution images from being obtained from deep regions of tissue.We review commontechniques,such as multiphoton microscopy(MPM)and optical coherence microscopy(OCM),for diffraction limited imaging beyond an imaging depth of 0.5 mm.Novel implementations havebeen emerging in recent years giving higher imaging speed,deeper penetration,and better imagequality.Focal modulation microscopy(FMM)is a novel method that combines confocal spatialfltering with focal modulation to reject out-of-focus background.FMM has demonstrated animaging depth comparable to those of MPM and OCM,near-real-time image acquisition,and thecapability for multiple contrast mechanisms.

Corneal alteration and pathogenesis in diabetes mellitus

The incidence of diabetes mellitus(DM) and its complications have increased considerably worldwide. Diabetic keratopathy is the major complication of the cornea characterized by delayed corneal wound healing, decreasing corneal epithelial sensitivity, and recurrent corneal ulcers. There is accumulating evidence that diabetic keratopathy is correlated with the hyperglycemic state. Different corneal components may produce different alterations under hyperglycemia. In addition, diabetic nerve alteration may become a novel biomarker of earlystage DM. Abnormalities of the corneal nerve plexus have been associated with diabetic inflammatory states. There is rapidly growing evidence based on investigations of diabetic corneal nerves through in vivo confocal microscopy. Understanding the molecular pathogenesis caused by hyperglycemia may assist in the identification of novel biomarkers, as well as therapeutic targets for early treatment. This review mainly summarizes recent findings on corneal alteration and pathogenesis in DM.

IN SITU OBSERVATION OF GROWTH BEHAVIOR AND MORPHOLOGY OF DELTA-FERRITE AS FUNCTION OF SOLIDIFICATION RATE IN AN AISI304 STAINLESS STEEL

It was presented the in situ observation of growth behavior and morphology of delta-ferrite as a function of solidification rate in an AISI304 stainless steel. The specimens have been solidified and observed using confocal scanning laser microscopy (CSLM). The δ-phase always appears like cells on the sample surface when critical supercooling occurs, during which the L→δ transformation starts. The solid-liquid (S-L) interface is found to be finger shaped and has no faceted shape. γ phase appears among δ grains due to partitioning of Ni into the melt during solidification, when solidification rate is higher. The mergence of observed δ cells is possible for the steel sample cooled at 7.5℃/min. The formation of dendrites can be observed on the free surface of the steel sample cooled at 150℃/min. The size of solidified delta grains decreases from 120 to 20-80μm, and the volume fraction of solidified austenite increases with increase in solidification rate from 7.5 to 150℃/min. The relation between the tip radius of δ cell and its growth rate is deduced, and the results agree with the experimental values.

Diabetic corneal neuropathy as a surrogate marker for diabetic peripheral neuropathy

Diabetic neuropathy is a prevalent microvascular complication of diabetes mellitus,affecting nerves in all parts of the body including corneal nerves and peripheral nervous system,leading to diabetic corneal neuropathy and diabetic peripheral neuropathy,respectively.Diabetic peripheral neuropathy is diagnosed in clinical practice using electrophysiological nerve conduction studies,clinical scoring,and skin biopsies.However,these diagnostic methods have limited sensitivity in detecting small-fiber disease,hence they do not accurately reflect the status of diabetic neuropathy.More recently,analysis of alterations in the corneal nerves has emerged as a promising surrogate marker for diabetic peripheral neuropathy.In this review,we will discuss the relationship between diabetic corneal neuropathy and diabetic peripheral neuropathy,elaborating on the foundational aspects of each:pathogenesis,clinical presentation,evaluation,and management.We will further discuss the relevance of diabetic corneal neuropathy in detecting the presence of diabetic peripheral neuropathy,particularly early diabetic peripheral neuropathy;the correlation between the severity of diabetic corneal neuropathy and that of diabetic peripheral neuropathy;and the role of diabetic corneal neuropathy in the stratification of complications of diabetic peripheral neuropathy.

Potential use of corneal confocal microscopy in the diagnosis of Parkinson's disease associated neuropathy

Parkinson's disease(PD)is a chronic,progressive neurodegenerative disease affecting about 2%-3% of population above the age of 65.In recent years,Parkinson's research has mainly focused on motor and non-motor symptoms while there are limited studies on neurodegeneration which is associated with balance problems and increased incidence of falls.Corneal confocal microscopy(CCM)is a real-time,non-invasive,in vivo ophthalmic imaging technique for quantifying nerve damage in peripheral neuropathies and central neurodegenerative disorders.CCM has shown significantly lower corneal nerve fiber density(CNFD)in patients with PD compared to healthy controls.Reduced CNFD is associated with decreased intraepidermal nerve fiber density in PD.This review provides an overview of the ability of CCM to detect nerve damage associated with PD.

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).

EFFECT OF COMPOSITION AND PROCESSING CONDITION ON MICROSTRUCTURAL PROPERTIES AND DURABILITY OF FLUOROPOLYMER/ACRYLIC BLENDS

Fluoropolymer blends have been widely used as binders for exterior coatings because of their excellent resistance to ultra-violet (UV) radiation as well as to many corrosive chemical agents.It is known that the fluorinated component usually has a lower glass transition temperature and easily crystallizes in the final structure depending upon the blend composition and sample annealing condition.We investigated the effect of blend composition and annealing process (slow and fast cooling) on the surface mor...

In vivo confocal microscopy: qualitative investigation of the conjunctival and corneal surface in open angle glaucomatous patients undergoing the XEN-Gel implant, trabeculectomy or medical therapy

Purpose:Assessing the quality of the ocular surface by in vivo scanning laser confocal microscopy(IVCM)in primary open angle glaucoma(POAG)patients treated by Xen 45 Gel Stent,medical therapy and trabeculectomy.Methods:Retrospective,single-center,single-masked,comparative study including 60 eyes of 30 patients(mean age 61.16±10 years)affected by POAG.Eyes were divided into 3 groups:Group 1 eyes underwent the Xen 45 Gel Stent procedure,Group 2 eyes were under medical therapy,Group 3 eyes were surgically treated by trabeculectomy.All patients underwent HRT II IVCM analysis of cornea,limbus,conjunctiva,sub-tenionian space and sclera.Results:The Xen 45 Gel stent,if properly positioned in the sub-conjunctival space preserves goblet cells and limits ocular surface inflammation.Regular corneal epithelial cells with micro-cysts,and normo-reflective sub-epithelial nerve plexus are documented by IVCM.In sub Tenon’s implants an alternative lamellar intra-scleral filtration is detectable.Combined surgical procedures show a noticeable number of inflammatory cells with rare micro-cysts.Post-trabeculectomy inflammatory reaction is more evident than Xen 45 Gel Stent associated surgical procedures,but less than medical therapy where a conspicuous presence of Langerhans cells,peri-neural infiltrates,marked loss of goblet cells and fibrosis is visible.Conclusion:Ocular surface inflammation was more notable in topical therapy than after trabeculectomy,which itself causes more inflammation than XEN Gel stents.

THE ROLES OF REACTION INHOMOGENEITY IN PHASE SEPARATION KINETICS AND MORPHOLOGY OF REACTIVE POLYMER BLENDS

The roles of reaction inhomogeneity in phase separation of polymer mixtures were described and summarized via two examples:photocross-link of polymer mixtures in the bulk state and photopolymerization of monomer in the liquid state. The reaction kinetics,the reaction-induced elastic strain and the phase separation kinetics were monitored respectively by UV-Vis spectroscopy,Mach-Zehnder interferometry and laser-scanning confocal microscopy.It was found that phase separation in the bulk state was strongly ...

Effect of the glyceryl monooleate-based lyotropic phases on skin permeation using in vitro diffusion and skin imaging

Glyceryl monooleate(GMO)is a polar lipid that can exist in various liquid crystalline phases in the presence of different amounts of water.It is regarded as a permeation enhancer due to its amphiphilic property.Various phases of GMO/solvent system containing sodium fluorescein were prepared to compare permeability using confocal laser scanning microscopy(CLSM).GMO was melted in a vial in a water bath heated to 45℃.Propylene glycol and hexanediol were homogeneously dissolved in the melted GMO.Sodium fluorescein in aqueous solution was diluted to various ratios and thoroughly mixed by an ultrasonic homogenizer.Each GMO/Solvent system with fluorescein was applied onto the epidermal side of excised pig skin and incubated overnight.CLSM was performed to observe how the GMO/solvent system in its different phases affect skin permeability.Cubic and lamellar phase formulations enhanced the fluorescein permeation through the stratum corneum.A solution system had the weakest permeability compared to the other two phases.Due to the amphiphilic nature of GMO,cubic and lamellar phases might reduce the barrier function of stratum corneum which was observed by CLSM as fluorescein accumulated in the dermis.Based on the results,the glyceryl monooleate lyotropic mixtures could be applied to enhance skin permeation in various topical and transdermal formulations.