Observations of human corneal epithelium by tandem scanning confocal microscope

We applied the tandem scanning confocal microscope (TSCM) to 30 healthy human corneas of 3 normal volunteers and 27 patients with cataract and retinal detachment to observe normal corneal epithelial cells in vivo. All corneas were normal under slit lamp microscopic examination. The superficial and basal epithelial cells close to the basal lamina in the central cornea were recorded on videotape and analyzed by a computer-assisted digitizer. The mean cell areas of superficial cells exposed at the surface and basal cells at the horizontal section were 624 ± 109 μm² and 66 ± 5 μm², respectively. The ratio of superficial to basal mean cell area was 11.0 ± 4.5. TSCM was thus useful in evaluating the relationship between superficial and basal cells in human corneal epithelium in vivo.     

Observations of human corneal epithelium by tandem scanning confocal microscope

We applied the tandem scanning confocal microscope (TSCM) to 30 healthy human corneas of 3 normal volunteers and 27 patients with cataract and retinal detachment to observe normal corneal epithelial cells in vivo. All corneas were normal under slit lamp microscopic examination. The superficial and basal epithelial cells close to the basal lamina in the central cornea were recorded on videotape and analyzed by a computer-assisted digitizer. The mean cell areas of superficial cells exposed at the surface and basal cells at the horizontal section were 624 ± 109 μm² and 66 ± 5 μm², respectively. The ratio of superficial to basal mean cell area was 11.0 ± 4.5. TSCM was thus useful in evaluating the relationship between superficial and basal cells in human corneal epithelium in vivo.     

Entropy of corneal nerve fibers distribution observed by laser scanning confocal microscopy: A noninvasive quantitative method to characterize the corneal innervation in Sjogren's syndrome patients

Sjogren's syndrome (SS) is a progressive autoimmune condition mainly affecting the salivary and lacrimal glands with an incidence of primary SS between 1/100 and 1/1,000. SS implies an alteration in the epithelium and subepithelium innervation, with consequent reduction of corneal sensitivity. It is necessary to have noninvasive quantitative methods to characterize the status of the corneal nerve fibers of the patients in order to choose and follow the best therapy. Entropy (information dimension) of the nerve corneal fibers distribution observed by confocal microscopy was evaluated in patients with primary SS (n = 30, 6 males, 24 females, 21–81 years), diagnosed by biopsy of salivary gland and blood tests and in sex‐ age‐matched healthy subjects (n = 12). Corneal nerve fiber density, Langerhans cell count, and cell density in the nerve plexus images were also evaluated. In selected patients salivary gland atrophy degree was also evaluated. Nerve corneal distribution observed by confocal microscopy is fractal. Entropy of the corneal nerve distribution statistically distinguishes between SS patients and healthy subjects: patients present a lower value of information dimension of the corneal nerve fibers distribution than healthy individuals (P < 0.001). Percentage of grouped cases classified by entropy according to the subjects (selected patients vs. healthy) showed a 100% sensitivity and 96% specificity, P < 0.0001 with a low value of coefficient of variation among the individuals (6–7 times lower than the other morphometric indexes). Entropy correlated with the severity of the disease (salivary gland atrophy degree, P < 0.01). Evaluation of entropy of the corneal nerve distribution observed by a laser confocal microscopy appears to quantitatively and noninvasively characterize an aspect of the SS patients in relation to the recognition of an impairment of their ocular surface, giving us for the first time a method to objectively and precisely characterize the corneal innervation status in the SS patients. Microsc. Res. Tech. 78:1069–1074, 2015. © 2015 Wiley Periodicals, Inc.     

Accessing osteocyte lacunar geometrical properties in human jaw bone on the submicron length scale using synchrotron radiation μCT

The architectural properties of the osteocyte cell network provide a valuable basis for understanding the mechanisms of bone remodelling, mineral homeostasis, ageing and pathologies. Recent advances in synchrotron microtomography enable unprecedented three‐dimensional imaging of both the bone lacunar network and the extracellular matrix. Here, we investigate the three‐dimensional morphological properties of osteocyte lacunae in human healthy and bisphosphonate‐related osteonecrotic jaw bone based on synchrotron X‐ray computed tomography images, with a spatial isotropic voxel size of 300 nm. Bisphosphonate‐related osteonecrosis of the jaw is a relatively new disease with increasing incidence, which remains poorly understood. A step forward in elucidating this malady is to assess whether, and how, the morphology of the osteocyte lacunar network is modified in the affected jaw tissue. We evaluate thousands of cell lacunae from five specimens of which three originate from patients diagnosed with bisphosphonate‐associated osteonecrosis. In this exploratory study, we report three‐dimensional quantitative results on lacunar volumes (296–502 μm³), shape (approximated by an ellipsoidal shape with principal axes a > b > c, such that a = 2.2b and a = 4c) and spatial distribution (i.e., 50% of the mineralized matrix volume is located within 12 μm to the closest lacunar boundary) at submicron resolution on such specimens. We observe that the average lacunar volumes of the bisphosphonate‐related osteonecrotic jaw specimens were within the range of volumes found in the two specimens originating from healthy donors and conclude that lacunar volumes are not the key element in the course of bisphosphonate‐related osteonecrotic jaw. In three out of five specimens we observe lacunar volume sizes in segmented osteons to be significantly different compared to lacunar volumes in the adjacent tissue regions. Furthermore, we quantify the number of lacunae containing small dense objects (on average 9%). In contrast to lacunar morphology we report the lacunar density (16 000–50 000 per mm³) to be different in jaw bone tissue compared to what has been reported in femoral sites.     

Counting cells with stereology: Random versus serial sectioning

Counts of cells and nuclei from sections provide information central to studying structural changes in cells, tissues, and organs. This study considers some of the practical problems associated with counting cells with the newer random and serial sectioning methods of stereology and tests the hypothesis that similar cell counts can be obtained with both random and serial sectioning methods. Using irregularly shaped nuclei from alveolar cells of the goat lung, we compared cell counts derived from random (electron microscopic) and serial sectioning (light microscopic) methods. The results showed that both sectioning methods gave similar cell counts (10⁷/cm³ of parenchyma) for type 1 epithelial cells (5.0 vs. 5.0; P=1.0), type 2 epithelial cells (8.6 vs. 9.8; P=0.42) and interstitial cells (34.6 vs. 33.4; P=0.64), provided that corrections were introduced for sectionrelated biases and that the nuclei of the random sectioning method were corrected for shape. We found counting biases of 5%–7% for nuclear shape and 16% for section compression. These observations support the hypothesis that similar cell counts can be obtained with random and serial sectioning, even when nuclei have irregular shapes.     

PIV as a method for quantifying root cell growth and particle displacement in confocal images

Particle image velocimetry (PIV) quantifies displacement of patches of pixels between successive images. We evaluated PIV as a tool for microscopists by measuring displacements of cells and of a surrounding granular medium in confocal laser scanning microscopy images of Arabidopsis thaliana roots labeled with cell‐membrane targeted green fluorescent protein. Excellent accuracy (e.g., displacement standard deviation <0.006 pixels) was obtained for root images that had undergone rigid digital translations of up to 40 pixels. Analysis of zoomed images showed that magnifications of up to 5% maintained good linear relations between PIV‐predicted and actual displacements (r² > 0.83). Root mean squared error for these distorted images was 0.4–1.1 pixels, increasing at higher magnification factors. Cell growth and rhizosphere deformation were tracked with good temporal (e.g., 1‐min interval) and spatial resolution, with PIV patches located on recognizable cell features being tracked more successfully. Appropriate choice of GFP‐label was important to decrease small‐scale biological noise due to intracellular motion. PIV of roots grown in stiff 2% versus 0.7% agar showed patterns of cell expansion consistent with physically impeded roots of other species. Roots in glass ballotini underwent rapid changes in growth direction on a timescale of minutes, associated with localized arching of ballotini. By tracking cell vertices, we monitored automatically cell length, width, and area every minute for 0.5 h for cells in different stages of development. In conclusion, PIV measured displacements successfully in images of living root cells and the external granular medium, revealing much potential for use by microscopists. Microsc. Res. Tech., 2009. © 2009 Wiley‐Liss, Inc.     

Electrolyte Jet Machining (EJM) of antibacterial surfaces

Electrolyte Jet Machining (EJM) has been performed on stainless steel surfaces with the aim of reducing bacterial retention through the generation of nanoscale surface morphology. Following initial EJM experiments aimed at investigating the influence of machining depth, machining speed and current density on the resulting surface roughness, three characteristic surfaces were produced with a current density of 10–18 A/cm² and a machining speed of 0.8–8 mm/s to obtain an arithmetic mean height (S_a) of 0.5–0.74 μm and a density of peaks (S_pd) of 0.25–1.26 μm⁻¹. Relatively large differences between the three surfaces in terms of S_pd allowed thorough investigation into the effects of surface feature size on bacterial retention to be performed. Reductions in the order of 90% compared to control samples were achieved for gram-positive Bacillus cereus and Staphylococcus aureus across the entire tested parameter range (S_pd = 0.25–1.26 μm⁻¹), while reductions in the order of 99% were achieved for gram-negative Escherichia coli and Pseudomonas aeruginosa for surfaces characterized by S_pd > 1 μm⁻¹. Not only do the results call attention to EJM as an innovative technology for producing antibacterial surfaces, they also highlight important differences in the behavior of gram-positive and gram-negative bacteria in relation to EJM-textured surfaces with nanoscale surface morphology.     

Functionalized magnetic nanoparticles attenuate cancer cells proliferation: Transmission electron microscopy analysis

The penetration and transportation of nanoparticles (NPs) inside the cancer cells is critical to study. In this article, cancer cells (HCT‐116) were treated with functionalized magnetic NPs for the period of 48 hr and studied their ultrastructure by transmission electron microscopy (TEM). The NPs‐treated cells were prepared by chemical fixation and sliced into electron‐transparent arbitrary sections (200 × 200 μm²) by ultramicrotome. Major events of NPs–cell interaction, such as penetration of NPs, encapsulation of NPs into the intracellular compartments, transportation of NPs, and NPs exit, were examined by TEM to understand the mechanism of cell death. The NPs showed the uniform spherical shape with broad size distribution (100–400 nm), while cells displayed irregular morphology with average diameter ~5 μm. Our results showed the successful penetration of NPs deep into the cell, encapsulation, transportation, and exocytosis. Furthermore, we tested the different concentrations (0, 1.5, 12.5, and 50 μg/ml) of NPs on cancer cells and evaluated the cell viability. Laser confocal microscopy and colorimetric analysis together demonstrated that the cell viability is a dose‐dependent phenomenon, where 50 μg/ml specimen showed the highest killing of cancer cells compared to other dosages.     

Analysis of microstructures and macrotextures for different apple cultivars based on parenchyma morphology

Fuji, Golden Delicious, and Jonagold parenchyma were imaged by confocal laser scanning microscopy to be extracted morphology characteristics, which were used to analyze the relationship with macrotexture of apples tested by penetration and compression. Before analyzing the relationship, the significantly different morphology parameters were reduced in dimensions by principal component analysis and were proved to be availably used for distinguishing the different apple cultivars. For compression results, cell did not absolutely determine the hardness in different apple cultivars, and the pore should also be taken into consideration. With the same size in cell feret diameter, the bigger the pore feret diameter was, the softer the apple became. If no difference existed in pore feret diameter size, the cultivar became harder with a narrower distribution in cell feret diameter. The texture parameters were compared with the roundness parameters in the same or inverse changing trends to explore the relationship. High correlations were found between the texture parameters (energy required in whole penetration (Wt), fracture force (Fp), crispness) and pore solidity (R² > 0.924, P < 0.001). Compactness of parenchyma played an important role in fruit texture. This research could provide the comprehension about relationship between microstructure and macrotexture of apple cultivars and morphological values for modeling apple parenchyma, contributing to numerical simulation for constitutive relation of fruit. Microsc. Res. Tech. 79:304–312, 2016. © 2016 Wiley Periodicals, Inc.     

Acute effects of splint immobilization of the forearm on in vivo microcirculation and histomorphology of the human skin

BACKGROUND: Splint immobilization of the forearm is often performed in clinical practice. Previous studies investigated the effect of immobilization on bone, cartilage, muscle, and tendon, however, the acute effects on human skin microcirculation and histomorphology remains elusive. METHODS: In 12 healthy, nonsmoking individuals (aged 29.7 ± 9.1 years) a randomly selected forearm was immobilized by splinting for 72 h, whereas the other forearm served as control. In vivo Reflectance‐Mode Confocal‐Microscopy (RMCM) was performed prior (baseline value) and postimmobilization to evaluate: quantitative blood cell flow; density of functional dermal capillaries; epidermal thickness; and granular cell size. RESULTS: At 72h forearm immobilization, quantitative blood cell flow was significantly reduced (42.86 ± 3.68 cells/min) compared to the control blood flow (53.11 ± 3.68 cells/min, P < 0.05) and dermal capillaries indicates less functional density (5.73 ± 0.63 capillaries/mm²) compared to the controls (7.04 ± 0.81 capillaries/mm², P < 0.05). Histometric assessment reveals significantly thinner epidermis following immobilization compared to the control site (40.02 ± 2.91 vs. 46.64±3.09 µm, P < 0.05). Granular cell size was significantly altered at 72 h splinting (730.1 ± 42.53 µm²) compared to the control cell size at 770.2 ± 38.21 µm². Comparison of baseline values of both forearms indicate statistically insignificance (P > 0.05) for each parameter. CONCLUSION: At 72 h splint immobilization, for the first time, significant adaptive mechanisms were evaluated on human skin microcirculation and histomorphology using in vivo RMCM. These adaptations may be considered as an incipient atrophy of the human skin. Long‐term effects of immobilization including the regenerative potential should be evaluated in further RMCM studies. Microsc. Res. Tech. 77:99–103, 2014. © 2013 Wiley Periodicals, Inc.     

Brief Note: Isolation,culture, characterization and optimization of human corneal stem cells

The effects of human versus mouse EGF on cell growth and culture duration were studied to optimize a human limbal stem cells culture method for therapeutical autologous transplantation. Limbal cells were obtained by trypsin digestion and transferred to a culture medium. The time needed to reach full confluence in culture was determined. Specific antibodies to corneal stem cell marker (P63) versus corneal epithelial differentiation marker (K3) were used for histochemical determinations. A high proportion of P63 positive cells (85± 4.6%), and a correspondingly low proportion K3 positive cells (15 ± 3.8%) indicated that most cultured cells remained undifferentiated and were considered as stem cells (mean ± SE, n=10). Cultures reached full confluency after 17.3 ± 1.2 days when the medium was supplemented with human EGF, while 21.7 ± 1.5 days were needed when the medium was supplemented with mouse EGF. The results showed that limbal stem cells proliferate more easily and reach to full confluency in a shorter time if the medium is supplemented with hEGF rather than with mEGF.     

角膜老年环检测方法研究

当人体脂类物质代谢发生异常时,会在角膜处形成角膜老年环。角膜老年环主要呈云雾环状覆盖在虹膜外环中,角膜老年环的检测可以作为一种辅助手段,对及时方便了解体内脂类代谢异常状况具有重要意义。针对目前方法中由于没有对角膜老年环边界精确检测而造成的鲁棒性不高,以及当存在K-F环时会发生检测错误的问题,提出了一种角膜老年环检测方法。首先,对其彩色图像进行分割与归一化;其次,利用基于梯度响应最优算法提取环状特征内外边界,通过定义的光照影响检测模型提高算法对光照的鲁棒性;最后,通过定义的宽度特征量化算子以及颜色特征量化算子完成角膜老年环的检测。实验结果表明,在采集的1 968幅图像中该方法能够达到96.3%的识别率,算法不仅不受光照影响而且当患者具有K-F环时,本文方法仍能够取得很好的检测效果。     

The use of confocal microscopy in evaluating corneal wound healing after excimer laser keratectomy

Corneal wound healing following excimer laser keratectomy is the major cause of regression of treatment results. The amount of anterior strorhal haze that develops may be influenced by topical medications. Over a period of 6 months, we followed 15 New Zealand white rabbit eyes that underwent excimer laser keratectomy with the VISX 193-nm ArF laser at a fluence of 150 mJ/cm² for a depth of 130 μm. Eyes were randomized to treatment with prednisolone acetate, diclofenac sodium (Voltaren), a combination of both, and a control group. Drops were administered four times a day for 1 week, two times a day for 3 weeks, and the drops were then tapered. All eyes were reepithelialized by 5 to 7 days. The tandem scanning confocal microscope (TSCM) was used to evaluate the corneal wound in vivo weekly for a month and monthly for 6 months. During the early postoperative period, the TSCM revealed significant anterior stromal keratocyte activation with cell elongation and the spindle-shaped appearance of fibroblasts in all groups. Collagenous stromal scarring was evident initially, then slowly decreased in all treatment groups. This study shows that TSCM is clinically useful for successive in vivo examinations of corneal wounds after excimer laser keratectomy and for comparing the effects of various topical medications.     

Effect of ethanol on the antimicrobial properties of chlorhexidine over oral biofilm

The aim of this study was to evaluate the effect of 95% ethanol irrigation, with 5 or 10 min of action, on the antibacterial properties of 2% chlorhexidine (CHX), on oral biofilm, evaluated with confocal laser scanning microscopy (CLSM). Oral biofilm development was induced in 80 sterilized bovine dentin blocks, distributed in two groups (5 or 10 min) and 4 subgroups, according to time and the solution used: Saline (SALINE5, SALINE10); Saline followed by CHX (SALINE/CHX5, SALINE/CHX10); Ethanol (ETHANOL5, ETHANOL10), Ethanol followed by CHX (ETHANOL/CHX5, ETHANOL/CHX10). The surface of the block was dyed with Live/Dead^® BacLight. Images from different areas were analyzed by BioImage L program. The total biovolum (µm³), biovolum of live cells (green), percentage of live cells of the thickness of the biofilm visualized in CLSM and on surface biofilm were evaluated. Total biovolum and biovolum of living cells showed similar results among the different groups (p > .05). The percentage of living cells in total thickness of the biofilm also was similar among the groups (p > .05), except ETHANOL5, SALINE/CHX10, ETHANOL10, and ETHANOL/CHX10 that showed lower percentage than SALINE5 (p < .05). The ETHANOL10 and ETHANOL/CHX10 also showed lower percentage of living cells than ETHANOL/CHX5 and SALINE10 (p < .05). In relation to biofilm surface, SALINE/CHX5, SALINE/CHX10, ETHANOL5, ETHANOL10, ETHANOL/CHX5, and ETHANOL/CHX10 showed a lower percentage of living cells percentage than SALINE5 and SALINE10 groups (p < .05). Therefore, ethanol has no effect on antimicrobial properties of 2% chlorhexidine, prior when used as endodontic irrigating solution.     

Near-field photothermal microspectroscopy for adult stem-cell identification and characterization

The identification of stem cells in adult tissue is a challenging problem in biomedicine. Currently, stem cells are identified by individual epitopes, which are generally tissue specific. The discovery of a stem-cell marker common to other adult tissue types could open avenues in the development of therapeutic stem-cell strategies. We report the use of the novel technique of Fourier transform infrared near-field photothermal microspectroscopy (FTIR–PTMS) for the characterization of stem cells, transit amplifying (TA) cells and terminally differentiated (TD) cells in the corneal epithelium. Principal component analysis (PCA) data demonstrate excellent discrimination of cell type by spectra. PCA in combination with linear discriminant analysis (PCA–LDA) shows that FTIR–PTMS very effectively discriminates between the three cell populations. Statistically significant differences above the 99% confidence level between IR spectra from stem cells and TA cells suggest that nucleic acid conformational changes are an important component of the differences between spectral data from the two cell types. FTIR–PTMS is a new addition to existing spectroscopy methods based on the concept of interfacing a conventional FTIR spectrometer with an atomic force microscope equipped with a near-field thermal sensing probe. FTIR-PTMS spectroscopy currently has spatial resolution that is similar to that of diffraction-limited optical detection FTIR spectroscopy techniques, but as a near-field probing technique has considerable potential for further improvement. Our work also suggests that FTIR–PTMS is potentially more sensitive than synchrotron radiation FTIR spectroscopy for some applications. Microspectroscopy techniques like FTIR–PTMS provide information about the entire molecular composition of cells, in contrast to epitope recognition that only considers the presence or absence of individual molecules. Our results with FTIR–PTMS on corneal stem cells are promising for the potential development of an IR spectral fingerprint for stem cells.     

In vivo observation of the corneal epithelium

Specular microscopy is an important addition to the ophthalmologist's diagnostic armamentarium. Using this technique, it has been found that normal corneal epithelial cells are polygonal, with no abnormal specular reflex. There are no spindle-shaped, large or small cells, as may be observed in a variety of pathologic conditions. Spindle-shaped cells are characteristic of the wound-healing process with cell migration. Large cells were observed not only in wound healing, but also in aphakic diabetic patients, extended-wear soft contact lens wearers, and individuals with keratoconus. These changes may result from depressed mitosis or inhibited sloughing of superficial cells. Morphometric analysis adds a quantitative dimension to epithelial cell analysis.     

In vivo observation of the corneal epithelium

Specular microscopy is an important addition to the ophthalmologist's diagnostic armamentarium. Using this technique, it has been found that normal corneal epithelial cells are polygonal, with no abnormal specular reflex. There are no spindle-shaped, large or small cells, as may be observed in a variety of pathologic conditions. Spindle-shaped cells are characteristic of the wound-healing process with cell migration. Large cells were observed not only in wound healing, but also in aphakic diabetic patients, extended-wear soft contact lens wearers, and individuals with keratoconus. These changes may result from depressed mitosis or inhibited sloughing of superficial cells. Morphometric analysis adds a quantitative dimension to epithelial cell analysis.     

Morphological changes in vero cells postinfection with dengue virus type‐2

Although no specific antiviral tablets or injections that can kill the dengue virus are currently available, adequate care and treatment could control its morbidity. Interaction of dengue virus to target cells could be an important feature for virus propagation. Ultrastructural analysis of this interaction was studied with vero cells. Vero cells were treated with Dengue virus type‐2 at different time intervals at multiplicity of infection (m.o.i) < 10, m.o.i > 10, and m.o.i = 100. It was found that m.o.i < 10 is best to study morphological changes. At an m.o.i > 10 apoptosis occurs and at m.o.i. = 100, cell necrosis occurs. While studying morphological changes, it was found that at 30 min postinfection cells have morphology very similar to that of the control cells although some have irregular outline and show cytoplasmic projections and intense cytoplasmic vacuolization. After 1–12 hours postinfection (h.p.i), the nuclei ran from normal looking to diffuse. Nuclear membrane begins to disintegrate. Some nucleoli are difficult to be seen. The cytoplasm appears as a mottled, lumps diffuse mass distributed throughout the cytosol, with dense lysosomes and myelin figures, also in the mitochondria. In later hours (24 h.p.i), the intranuclear euchromatin is dispersed and heterochromatin forms peripheral clumps. The cytoplasmic processes are short and few in numbers. A proportion of damaged mitochondria with disrupted cristae appear, suggesting that dengue virus may induce mitochondrial dysfunction and nucleus and mitochondria may be the primary organelles helping in dissemination of virus. Microsc. Res. Tech. 2011. © 2010 Wiley‐Liss, Inc.     

Biopolymer wear screening rig validated to ASTM F732-00 and against clinical data

Abstract

The ASTM standard F732-00, 'standard test method for wear testing of polymeric materials used in total joint prostheses' offers several pieces of data against which a test rig for such biopolymers can be validated. These conditions include a linear wear in the range from 0˙07 to 0˙2 mm/million cycles for ultrahigh molecular weight polyethylene (UHMWPE), and a wear rate ranking of poly tetra fluoro ethylene (PTFE)>>polyacetal≥UHMWPE. Using a modified pin on plate wear test rig, these three biopolymers were tested under a 40 N load against polished stainless steel plates while using dilute bovine serum heated to 37°C as the lubricant. When subject to multidirectional motion, mean wear factors of 37×10^–6 mm³ Nm^–1 for PTFE, 3˙8×10^–6 mm³ Nm^–1 for polyacetal and 1˙1×10^–6 mm³ Nm^–1 for UHMWPE were obtained. Therefore the wear ranking recommended by the ASTM standard was achieved. When the corrected mean weight loss of the UHMWPE test pins was converted to a linear wear rate, a value of 0˙12 mm/million cycles was obtained. Therefore the ASTM recommended target for linear wear rate was met. When the wear factors from the test materials were compared with clinical wear factors of 37×10^–6 mm³ Nm^–1 for PTFE, 3˙5×10^–6 mm³ Nm^–1 for polyacetal and 0˙95– 1˙45×10^{– 6} mm³ Nm^–1 for UHMWPE, good correspondence between in vivo and in vitro wear factors was obtained, further validating the modified pin on plate wear test rig.     

基于多尺度颜色替换的角膜老年环分割

角膜老年环是一种在角膜边缘形成的白色环状改变,主要由于人体脂类代谢异常而产生。通过图像分析的方法对角膜老年环进行检测可以方便、及时帮助人们发现身体脂类代谢异常状况。自然睁开状态下获取的彩色图像,角膜老年环经常被眼睑随机遮挡,而且被光斑、血管等因素的干扰。为了提高算法鲁棒性,减少由于眼睑的随机遮挡造成的定位失误,提出了一种基于多尺度颜色替换的角膜老年环分割方法。首先对图像进行量化;其次,在不同尺度模板下对图像按照本文定义的颜色替换策略进行处理,并最终实现目标分割。实验结果表明,在采集的1 968幅图像中该方法能够达到97.0%的分割正确率,所用算法具有较高的鲁棒性。