Interface quality of endothelial keratoplasty buttons obtained with optimised femtosecond laser settings.

AIM: To optimise interfaces of endothelial buttons created with femtosecond (FS) lasers. SETTING: Department of Ophthalmology, Hôtel-Dieu Hospital, Paris, France. METHODS: Forty-two corneas were divided into five groups of various cutting patterns and a control group of 100 µm laser in situ keratomileusis flap creation. A single path full lamellar cut (500 µm) was applied to groups 1 and 2. The same full lamellar cut was applied twice to groups 3 and 4. Two successive lamellar cuts were performed in group 5 (350 and 150 µm). 60 kHz and 150 kHz were used respectively in groups 1, 3, 5, 6 and 2, 4. In each group, different laser settings were tested to obtain the best interface quality while delivering minimal energy to the stroma. The quality of stromal interfaces from created endothelial lenticules was observed using a scanning electron microscope. RESULTS: Stromal adherences persisted after both the single- and double-path procedure, creating central irregularities on the endothelial lenticule. Among all groups and settings tested, the double-layer pattern (group 5) with FS full lamellar cut parameters set for diameter (mm), depth (µm), energy (µJ) and spot size/step (µm) respectively on 9.0 mm, 350 µm, 2.1 µJ, 4:4 µm and 8.3 mm, 150 µm, 0.9 µJ, 4:4 µm created the smoothest interfaces with the best reproducibility. CONCLUSIONS: Buttons for endothelial keratoplasty can be created with FS laser with a stromal interface quality comparable with that of refractive surgery.

Genotype-phenotype correlations of TGFBI p.Leu509Pro, p.Leu509Arg, p.Val613Gly, and the allelic association of p.Met502Val-p.Arg555Gln mutations.

PURPOSE: Investigate the genotype-phenotype correlations for five TGFBI (transforming growth factor, beta-induced) mutations including one novel pathogenic variant and one complex allele affecting the fourth FAS1 domain of keratoepithelin, and their potential effects on the protein's structure. METHODS: Three unrelated families were clinically diagnosed with lattice corneal dystrophy (CD) and one with an unclassified CD of Bowman's layer. Mutations in the TGFBI gene were detected by direct sequencing, and the functional impact of each variant was predicted using in silico algorithms. Corneal phenotypes, including histological examinations, were compared with the literature data. Furthermore, molecular modeling studies of these mutations were performed. RESULTS: Two distinct missense mutations affecting the same residue at position 509 of keratoepithelin: p.Leu509Pro (c.1526T>C) and p.Leu509Arg (c.1526T>G) were found to be associated with a lattice-type CD. The novel p.Val613Gly (c.1828T>G) TGFBI mutation was found in a sporadic case of an Algerian individual affected by lattice CD. Finally, the Bowman's layer CD was linked to the association in cis of the p.Met502Val and p.Arg555Gln variants, leading to the reclassification of this CD as atypical Thiel-Behnke CD. Structural modeling of these TGFBI mutations argues in favor of these mutations being responsible for instability and/or incorrect folding of keratoepithelin, predictions that are compatible with the clinical diagnoses. CONCLUSIONS: Description of a novel TGFBI mutation and a complex TGFBI allele further extends the mutational spectrum of TGFBI. Moreover, we show convincing evidence that TGFBI mutations affecting Leu509 are linked to the lattice phenotype in two unrelated French families, contrasting with findings previously reported. The p.Leu509Pro was reported to be associated with both amyloid and non-amyloid aggregates, whereas p.Leu509Arg has been described as being responsible for Epithelial Basement Membrane Dystrophy (EBMD).

Carotid artery dissection revealed by an oculosympathetic spasm.

Internal carotid dissection can be responsible for stroke and lead to severe neurological and functional complications. Thus, it must be diagnosed and treated with heparin as soon as possible. Horner syndrome is one of the most usual manifestations of internal carotid dissection. We report the case of a patient who presented with a unilateral non-reactive enlargement of the right pupil that did not last longer than 30 s. As a carotid dissection was not recognized from this atypical symptomatology, magnetic resonance angiography was performed only a few days later when Horner syndrome occurred. It disclosed a dissection of the internal carotid artery ipsilateral from its origin. The evolution and the duration of the pupil involvement suggest that the initial episode of mydriasis was caused by an oculosympathetic spasm, a rare form of sympathetic dysfunction that can be observed when the sympathetic nerve or the pericarotid plexus is irritated. It is important to recognize this oculosympathetic spasm because it has equal value as Horner syndrome for the diagnosis of internal carotid dissection.

Average 3-dimensional models for the comparison of Orbscan II and Pentacam pachymetry maps in normal corneas.

PURPOSE: To assess the reliability of Orbscan (Bausch & Lomb, Salt Lake City, UT) and Pentacam (Oculus, Wetzlar, Germany) central corneal thickness (CCT) and peripheral corneal thickness (PCT) measurements based on 2 methodologies. DESIGN: Evaluation of a diagnostic technology. PARTICIPANTS: Thirty healthy volunteers were recruited prospectively at the Department of Ophthalmology of the Hôtel-Dieu Hospital, Paris, France. METHODS: Central corneal thickness and PCT were assessed, using ultrasound pachymetry (USP) as the gold standard. Two methodologies were used: (1) the traditional analysis of pachymetry data from 1 central and 8 peripheral reference positions on the cornea, and (2) a 3-dimensional (3-D) analysis based on average corneal pachymetry maps constructed for each system (Orbscan, Pentacam, and USP), each operator (operators 1 and 2), and each visit (visits A and B). MAIN OUTCOME MEASURES: Repeatability, intersystem reproducibility, interoperator reproducibility, reproducibility over time, and accuracy of Orbscan and Pentacam CCT and PCT measurements. Distribution and statistical significance of the differences between 3-D average maps. RESULTS: Repeatability (Orbscan intraclass correlation coefficients [ICCs], 0.967-0.992; Pentacam ICCs, 0.986-0.997), interoperator reproducibility, and reproducibility over time (ICCs, 0.976-0.997) were excellent to almost perfect for both systems. Intersystem agreement was almost perfect for CCT (ICC, 0.980), but less strong for PCT (ICCs, 0.928-0.979). Despite a good to excellent agreement between the optical systems and USP (ICCs, 0.608-0.958), USP CCT readings were thicker (mean difference, up to 15.2 microm; P<0.05), and USP PCT readings were thinner (P<0.05). Orbscan and Pentacam average maps allowed comprehensive interpretation of differences between populations according to the magnitude, distribution, and statistical significance, minimizing the risk of giving excessive weight to few data measured at specific locations on the cornea. CONCLUSIONS: Both methodologies showed that Orbscan and Pentacam CCT readings are interchangeable, whereas caution should be used for PCT readings. Interchangeability with USP measurements also was shown to be limited. The high repeatability, interoperator reproducibility, reproducibility over time, the extent of the information generated by a single capture, and the noncontact nature of the Orbscan and Pentacam all suggest that optical systems eventually may replace USP as the gold standard for corneal pachymetry.

Severe Keratitis Caused by Pseudomonas aeruginosa Successfully Treated with Ceftazidime Associated with Acetazolamide.

Purpose. To report a case of microbial keratitis caused by Pseudomonas aeruginosa treated with a combination of acetazolamide and ceftazidime. Methods. Case report. Results. We report the case of a 17-year-old contact lens-wearing female who developed severe keratitis due to Pseudomonas aeruginosa temporarily healed with topical fortified antibiotic eye drops. After few days, the patient relapsed, and topical and intravenous ceftazidime were added. Concomitantly, oral administration of acetazolamide was prescribed. This carbonic anhydrase inhibitor was added to the antibiotic regimen in order to decrease the anterior chamber pH, and then, the ceftazidime ionization. By lowering the state of ionization of the antibiotic in the aqueous humor, its concentration was increased. This was confirmed by an improvement of the patient within few days and a rapid eradication of the infection. Conclusion. This is the first reported case of keratitis caused by P. aeruginosa successfully treated using acetazolamide as an enhancer of ceftazidime effectiveness.

Displacement of retained subfoveal perfluorocarbon liquid after vitreoretinal surgery.

OBJECTIVE: To report a novel surgical procedure to displace retained subfoveal perfluorocarbon liquid (PFCL). METHODS: Retrospective cases series. Three patients had retained subfoveal droplets after PFCL was used in vitrectomy repair of retinal detachment. In each case, submacular PFCL was displaced to the subretinal space in the inferior periphery. A retinal detachment at the posterior pole and the inferior periphery was created by injecting balanced salt solution through a retinal puncture near the inferotemporal vessels. Fluid-air exchange was performed, followed by short-term postoperative upright head positioning. RESULTS: In all cases, PFCL droplets were successfully displaced toward the inferior periphery, with good visual recovery. CONCLUSION: This procedure seems to be safe and is an alternative to direct aspiration of PFCL through a juxtafoveal retinotomy, which risks damage to the foveal region.

Nitrosative stress and corneal transplant endothelial cell death during acute graft rejection.

BACKGROUND: Nitrosative stress takes place in endothelial cells (EC) during corneal acute graft rejection. The purpose of this study was to evaluate the potential role of peroxynitrite on corneal EC death. METHODS: The effect of peroxynitrite was evaluated in vivo. Fifty, 250, and 500 microM in 1.5 microL of the natural or denatured peroxynitrite in 50 microM NaOH, 50 microM NaOH alone, or balanced salt solution were injected into the anterior chamber of rat eyes (n=3/group). Corneal toxic signs after injection were assessed by slit-lamp, in vivo confocal imaging, pachymetry, and EC count. The effect of peroxynitrite was also evaluated on nitrotyrosine and leucocyte elastase inhibitor/LDNase II immunohistochemistry. Human corneas were incubated with peroxynitrite and the effect on EC viability was evaluated. A specific inducible nitric oxide synthase inhibitor (iNOS) was administered systemically in rats undergoing allogeneic corneal graft rejection and the effect on EC was evaluated by EC count. RESULTS: Rat eyes receiving as little as 50 microM peroxynitrite showed a specific dose-dependent toxicity on EC. We observed an intense nitrotyrosine staining of human and rat EC exposed to peroxynitrite associated with leucocyte elastase inhibitor nuclear translocation, a noncaspase dependent apoptosis reaction. Specific inhibition of iNOS generation prevented EC death and enhanced EC survival of the grafted corneas. However, inhibition of iNOS did not have a significant influence on the incidence of graft rejection. CONCLUSION: Nitrosative stress during acute corneal graft rejection in rat eyes induces a noncaspase dependent apoptotic death in EC. Inhibition of nitric oxide production during the corneal graft rejection has protective effects on the corneal EC survival.

H244R VSX1 is associated with selective cone ON bipolar cell dysfunction and macular degeneration in a PPCD family.

PURPOSE: To elucidate the retinal dysfunction and the molecular basis of posterior polymorphous corneal dystrophy (PPCD) associated with macular dystrophy, both inherited in a dominant manner through a three-generation family. METHODS: Ophthalmologic examinations including slit lamp examination, visual acuity tests, fundus visualization by scanning laser ophthalmoscopy, fluorescein angiography, color vision tests, electro-oculography, photopic and scotopic electroretinography (ERG) according to the International Society for Clinical Electrophysiology of Vision (ISCEV) protocols, and oscillatory potential (OP) recordings were conducted on affected family members. Corneal button from one affected patient was examined by transmission electron microscopy. All exons and intron-exon boundaries of the VSX1 and the COL8A2 genes were amplified by polymerase chain reaction and sequenced. RESULTS: The presence of endothelial cells that have epithelial-like features with multiple layers, desmosomal junctions, and microvillous projections supports the diagnosis of PPCD. Sequence analysis indicated that the H244R variant in the VSX1 segregated with corneal and macular disease phenotypes in this family. Electrophysiologic studies indicated normal scotopic ERG findings, decreased amplitude of the photopic b-wave, photopic OP2 and OP3 barely recordable with a preserved OP4 amplitude, and variably decreased 30-Hz flicker amplitude. CONCLUSIONS: The human VSX1 is required for cone ON bipolar cell function but not for rod and cone OFF bipolar cells, giving a unique example of such a selective heritable retinal defect in humans. Furthermore, the authors provide the first clinical support for a new alternative role of VSX1 in cone biology, probably similar to that proposed for its goldfish ortholog during retinal differentiation.

Traumatic globe rupture following penetrating keratoplasty.

PURPOSE: To evaluate the mechanism, clinical characteristics, management and visual outcome of ocular trauma following penetrating keratoplasty (PK). METHODS: Twenty-six patients (13 men, 13 women) who suffered surgical wound dehiscence after PK because of ocular blunt trauma between 1994 and 2001 were included in this retrospective study. Graft dehiscence was managed with primary wound closure in all patients. Visual acuity, intraocular pressure, and funduscopy were evaluated in the follow-up. RESULTS: The mean age at trauma was 50+/-24 years (range 9-88 years). Mean interval between transplantation and trauma was 45.5+/-64 months (range 1 month to 20 years). Globe rupture occurred at the graft-host junction in all patients. Nine of 13 phakic eyes (69.2%) presented lens expulsion. Eight of nine pseudophakic eyes (88.9%) had lost their implants. Retinal detachment occurred in seven eyes (27%) within 3 months following the trauma. Five patients (19.2%) underwent vitreous surgery for posterior segment damage. Two eyes (7.7%) were regrafted. At the last examination, only seven eyes (27%) had visual acuity of 20/200 or better. CONCLUSIONS: Traumatic wound dehiscence may occur, and the prognosis is poor after the injury. Globe rupture at the graft-host junction showed persistence of wound weakness even a long time after PK. Prevention of ocular trauma should be performed following PK.

Cornea graft endothelial cells undergo apoptosis by way of an alternate (caspase-independent) pathway.

PURPOSE: To look for apoptosis pathways involved in corneal endothelial cell death during acute graft rejection and to evaluate the potential role of nitric oxide in this process. MATERIALS AND METHODS: Corneal buttons from Brown-Norway rats were transplanted into Lewis rat corneas. At different time intervals after transplantation, apoptosis was assessed by diamino-2-phenylindol staining and annexin-V binding on flat-mount corneas, and by terminal transferase dUTP nick end labeling (TUNEL), caspase-3 dependent and leukocyte elastase inhibitor (LEI)/LDNase II caspase-independent pathways on sections. Inducible nitric oxide synthase (NOS-II) expression and the presence of nitrotyrosine were assayed by immunohistochemistry. RESULTS: Graft endothelial cells demonstrated nuclear fragmentation and LEI nuclear translocation, annexin-V binding, and membranes bleb formation. Apoptosis associated with caspase-3 activity or TUNEL-positive reaction was not observed at any time either in the graft or in the recipient corneal endothelial cells. During 14 days posttransplantation, the recipient corneal endothelial cells remained unaltered and their number unchanged in all studied corneas. NOS-II was expressed in infiltrating cells present within the graft. This expression was closely associated with the presence of nitrotyrosine in endothelial and infiltrating cells. CONCLUSION: During the time course of corneal graft rejection, graft endothelial cells undergo apoptosis. Apoptosis is caspase 3 independent and TUNEL negative and is, probably, carried out by an alternative pathway driven by an LEI/L-Dnase II. Peroxynitrite formation may be an additional mechanism for cell toxicity and programmed cell death of the graft endothelial cells during the rejection process in this model.

Recurrence of keratoconus characteristics: a clinical and histologic follow-up analysis of donor grafts.

PURPOSE: To report on clinical corneal topography, histopathologic analysis, and fine structure findings in failed grafts after penetrating keratoplasty (PK) for keratoconus (KC). DESIGN: Retrospective, consecutive, interventional case series with histologic and clinical correlation. PARTICIPANTS: Twelve corneal buttons were obtained from consecutive patients undergoing repeated PK 10 to 28 years after the initial PK for KC. The indication for regrafting was endothelial deficiency in seven cases, irreversible immune graft rejection in two cases, and corneal ectasia in three cases. METHODS: Removed corneal buttons were examined by light and transmission electron microscopy. A potential correlation between the clinical and videokeratoscopic findings and the microscopic structural observations was analyzed. RESULTS: Preoperative simulated keratometry measured by TMS-1 (Tomey, New York, NY) or EyeSys CAS (EyeSys Technology, Houston, TX) ranged from 49.8 to 66.1 diopters. A pattern compatible with KC characteristics was observed in all cases. Fine structure analysis revealed Bowman's layer disruption or folds and stromal deposits in all corneal buttons. However, central corneal thinning was not present in any of the removed buttons. CONCLUSIONS: Structure changes compatible with the diagnosis of KC were observed in all donor buttons many years after PK on KC recipients. Recurrence of the KC characteristics may result from graft repopulation by recipients' keratocytes, aging of the grafted tissue, or both.

Late swelling and removal of Miragel buckles: a comparison with silicone indentations.

BACKGROUND: To describe the indications for removal of Miragel episcleral buckles and to compare them with removed silicone sponges and solid silicone indentations. METHODS: Retrospective analysis of the charts of patients successfully treated for retinal detachment in whom the episcleral buckle was removed after surgery, with a minimal follow-up of 6 months. RESULTS: In a series of 90 eyes of 90 patients, including 38 cases of Miragel elements, 25 cases of solid silicone, and 27 cases of silicone sponge removal, the duration of indentation before removal was significantly longer with Miragel (91.9 months) than with solid silicone (10.6 months) and silicone sponges (18.6 months). In eyes treated with Miragel elements, swelling of the material with progressive limitation of ocular motility and protrusion of the buckle beneath the eyelids indicated its removal in 34 (89.5%) eyes. Infection of the buckle and erosion of the conjunctiva with an exposed indentation were significantly more common with silicone buckles. Overall, scleral perforation occurred in four (4.4%) eyes, and retinal redetachment occurred in eight (8.8%) eyes. CONCLUSION: Late swelling of Miragel buckling elements represents a common indication for buckle removal, significantly different from silicone indentations.

Intrastromal corneal ring segments and corneal anterior stromal necrosis.

Poly(methyl methacrylate) refractive intracorneal ring segments (ICRS) can be removed for a refractive miscorrection or for early complications after implantation. We report the first case of a woman who experienced anterior stromal necrosis 5 years after an ICRS surgical procedure.

Truncating mutations in the carbohydrate sulfotransferase 6 gene (CHST6) result in macular corneal dystrophy.

PURPOSE: Identification of mutations in the CHST6 gene in 15 patients from 11 unrelated families affected with recessive macular corneal dystrophy (MCD). METHODS: Genomic DNA was extracted from peripheral blood leukocytes of the affected patients and their healthy family members, and the mutational status of the CHST6 gene was determined for each patient by a PCR-sequencing approach. Serum concentrations of antigenic keratan sulfate for each proband were determined by ELISA. RESULTS: ELISA indicated that all affected patients, except one, were of MCD type I or IA. Fourteen distinct mutations were identified within the CHST6 coding region: 2 nonsense, 2 frameshift, and 10 missense. Of these, 12 were novel, and a nonsense mutation in the homozygous state is reported for the first time. CONCLUSIONS: These molecular results in French patients with MCD combined with those reported in previous studies indicated CHST6 mutational heterogeneity. The characterization herein of nonsense mutations is in keeping with the fact that MCD results from loss of function of the CHST6 protein product.

Phototherapeutic keratectomy for BIGH3-linked corneal dystrophy recurring after penetrating keratoplasty.

PURPOSE: To determine visual results and report side effects and complications after phototherapeutic keratectomy (PTK) for BIGH3-linked corneal dystrophy recurring after penetrating keratoplasty. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Forty-two excimer laser PTK procedures were performed in 42 eyes of 29 patients with BIGH3-linked corneal dystrophies. Genetic status of all patients was determined and allowed us to assess an unambiguous diagnosis. Preoperative diagnoses included LCDIIIA/A546T (1 eye), R124 l+DT125-DE126 (4 eyes), GICD/R555W (14 eyes), LCDI/R124C (6 eyes), SGD/R124 l (16 eyes), and CDBII/R555Q (1 eye). INTERVENTION: Two excimer lasers (Summit Excimed UV 200, Summit Technology, Waltham, MA and Nidek EC 5000, Nidek, Inc., Gamagori, Japan) were used to perform all PTKs. Indications for performing PTK after a graft were severe decrease of the best-corrected visual acuity (BCVA) related to recurrent corneal deposits and/or painful recurrent epithelial erosions. MAIN OUTCOME MEASURES: Preoperative and postoperative BCVA were analyzed, significant recurrences after treatment were noted, and postoperative complications were recorded. RESULTS: Mean preoperative BCVA was 0.2 +/- 0.12 in the decimal chart, mean postoperative BCVA was 0.52 +/- 0.16 with a mean follow-up of 3.13 +/- 1.77 years (range, 0.3-6.65 years). Visual acuity was significantly improved after surgery (P < 0.05). The magnitude of the change in visual acuity was dependent on the mutation (P < 0.001). Seven symptomatic recurrences were observed. One regressive graft rejection and 4 cases of severe postoperative haze were observed. No other complications were noted. CONCLUSIONS: PTK is a simple, safe, and efficient technique for the treatment of recurrent corneal dystrophies; in many cases it prevents or delays the major incumbent problems of repeated grafting.

Cryoextraction of episcleral Miragel buckle elements: a new technique to reduce fragmentation.

A step-by-step description of a surgical technique for episcleral Miragel buckle removal is reported. After dissection of the conjunctiva and the capsule surrounding the Miragel element, the extremity of the buckle is pulled out with a cryoprobe. The removal of Miragel episcleral buckle with the cryoprobe is a safe and effective technique with a low fragmentation rate and a reduction of the surgical risk related to the use of sharp forceps near a possibly weakened sclera. Procedure time seems also to be reduced.

Clinical outcome of eight BIGH3-linked corneal dystrophies.

OBJECTIVE: To determine whether the mutational pattern of BIGH3-linked corneal dystrophies (CDs) can accurately predict the clinical course of the disease and be helpful in planning adequate surgical treatment. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Chart review of 73 patients (110 eyes) with recently confirmed BIGH3 mutations who underwent a penetrating keratoplasty (PK) from 1978 through 1999. Diagnoses included Thiel-Benhke CD (TBCD/R555Q) (13 eyes), classic granular CD (CGCD/R555W) (28 eyes), superficial variant of granular dystrophy (SVGD/R124 l) (27 eyes), lattice CD type I (LCDI/R124C) (20 eyes), Avellino CD (ACD/R124H) (2 eyes), H626R-lattice dystrophy (LCD/H626R) (6 eyes), and two novel dystrophies: a French variant of granular dystrophy (FVGD/R124 l+DT125-DE126) (9 eyes) and a French lattice CD type IIIA (LCDIIIA/A546T) (5 eyes). METHODS: The mutation of the BIGH3 gene was characterized for all patients. Clinical data were reviewed for each patient, and included age at first PK and elapsed time before significant recurrence (as defined by a severe decrease in best-corrected visual acuity related to recurrent deposits in the graft). MAIN OUTCOME MEASURES: Mean age at first PK and delay before a significant recurrence. RESULTS: Mutational pattern was highly correlated with the clinical course of each dystrophy. According to the genetic mutation, two groups with different prognosis were identified. Group 1 was defined by the presence of the FVGD/R124 l+DT125-DE126 and SVGD/R124 l mutations and was characterized by the early need for treatment and early recurrence of deposits. Group 2 was molecularly defined by the presence of any of the following mutations: LCDI/R124C, CGCD/R555W, LCDIIIA/A546T, TBCD/R555Q, and LCD/H626R. In group 2, mean age at first treatment was older, and delay before a significant recurrence was longer as compared with group 1 (P = 0.0001). CONCLUSIONS: These results demonstrate that there is a direct correlation between the molecular defect and the clinical course of BIGH3-linked CDs. They also indicate that molecular characterization of the genetic defect will help predict and design adequate surgical treatment for patients with ambiguous clinical diagnosis.

Estudo em microscopia confocal do epitélio corneano do coelho e do homem utilizando objetivas com diferentes aberturas numéricas / Confocal microscopic study of rabbit and human corneal epithelia using two objetives with different numerical aperture

Objetivo: Este estudo prospectivo compara as imagens de microscopia confocal do epitélio corneano do coelho e do homem, obtidas através de 2 objetivas com aberturas numéricas (AN) diferentes. Métodos: Dez olhos de coelhos foram enucleados e fixados através de um suporte pneumático para garantir o melhor desempenho de cada objetiva. Cinco pacientes normais foram selecionados após consentimento. Os olhos de coelhos e dos pacientes foram previamente examinados na lâmpada de fenda. O exame de microscopia confocal (Tomey, Erlangen-Tennenlohe, Alemanha) foi realizado com as objetivas achroplan 40x/AN = 63x/AN = 0,9 (Zeiss Oberkochen, Alemanha). Imagens selecionadas do epitélio corneano foram avaliadas qualitativamente com relaçäo ao tamanho, forma e refl tividade das células. Resultados: As células no epitélio superficial dos coelhos e dos pacientes, previamente à descamaçäo, tiveram uma refletividade maior que as células adjacentes. Este aspecto foi claramente observado somente com a objetiva 63x/AN = 0,9. As camadas basal e intermediária do epitélio em coelhos foram visualizadas somente através desta objetiva. Estas camadas nos pacientes tornaram-se mais nítidas com a objetiva de abertura numérica maior (63x/AN = 0,9). Conclusäo: Uma objetiva de abertura numérica elevada p oduz melhor resoluçäo dos cortes ópticos, facilitando a análise as camadas do epitélio no coelho e no homem