Optical coherence tomography and confocal laser scanning microscopy as non-invasive tools in the diagnosis of sinonasal inverted papilloma: a pilot study.

Optical coherence tomography (OCT) and confocal laser scanning microscopy (CLSM) are light-based imaging techniques that allow for a visualization of microscopic tissue properties in vivo. Our study was to examine whether they allow for differentiation of inverted papilloma (IP) from nasal polyps (NP). Five cases of IP and NP, respectively, were investigated intraoperatively with OCT and CLSM. Biopsies were taken of the investigated area and were analyzed ex vivo with OCT and CLSM and then underwent HE-staining for standard light microscopy. On OCT images, IP showed the characteristic inverted character of the epithelium, that was thicker with a high degree of variability of thickness compared to the thin and homogenous epithelium of NP. In addition, the characteristic stromal edema of NP could be visualized. On CLSM images, the typical epithelial invaginations of IP appeared as crypts, while in NP the highly organized cylindric epithelium could be visualized. In vivo, OCT acquired images of sufficient quality to visualize these characteristics, while CLSM did not. Our study demonstrates that OCT and CLSM can distinguish IP from NP. Further technical development is required to apply the techniques clinically to guide intranasal biopsies or even to make them dispensable.

[Biomechanical investigations on accommodation of the eye]. / Biomechanische Untersuchungen zur Akkommodation des Auges.

BACKGROUND: The biomechanics of accommodation are of particular interest in terms of the causes of presbyopia and the function of intraocular lenses. OBJECTIVE: The aim of the present article is to model the mechanism of accommodation in detail. MATERIALS AND METHODS: The state of the art of applying biomechanical models to accommodation is presented, which enables the accommodation process to be understood. RESULTS AND CONCLUSION: The established models, which are based on the Helmholtz theory, can explain the accommodation process in a plausible manner. These models thereby also enable further investigations on the genesis of presbyopia as well as on the development of accommodative intraocular lenses and implants. However, measurements are always necessary to compare the simulation results with reality, and to provide input and material data as well as geometric dimensions of components of the eye.

[In vivo imaging of the corneal nerve plexus : From single image to large scale map]. / In-vivo-Bildgebung des kornealen Nervenplexus : Vom Einzelbild zur großflächigen Darstellung.

The sub-basal nerve plexus (SNP) of the cornea provides the possibility of in vivo and non-invasive examination of peripheral nerve structures by corneal confocal microscopy (CCM). Thus morphological alterations of the SNP can be directly detected and quantified. A single CCM image is insufficient for a well-founded diagnosis because of the inhomogeneous distribution of the nerve fibers; therefore, there is a demand for techniques for large area imaging of the SNP. This article provides an overview of published approaches to the problem. Current developmental work at the Karlsruhe Institute of Technology and the University of Rostock Eye Clinic is expected to lead to a simplified handling of the technology and a further improvement in the image quality.

[Morphometric characterization of the subbasal nerve plexus : Detection and analysis of networks of nerve fibers]. / Morphometrische Charakterisierung des subbasalen Nervenplexus : Detektion und Analyse von Nervenfasernetzwerken.

Confocal laser scanning microscopy is a versatile tool in medical research and enables noninvasive in vivo imaging of the corneal subbasal nerve plexus. The aim of this work is to provide a structured overview about the detection and quantification of nerve fibers of the subbasal nerve plexus from images acquired by confocal laser scanning microscopy. Relevant steps are explained and potential factors influencing the quality of the results are pointed out. Information obtained from the quantification of subbasal nerve fiber structure can be potentially used as clinical parameters in the context of diagnostics and therapy control of diabetic neuropathy.

Femtosecond laser cutting of human corneas for the subbasal nerve plexus evaluation.

Assessment of various morphological parameters of the corneal subbasal nerve plexus is a valuable method of documenting the structural and presumably functional integrity of the corneal innervation in health and disease. The aim of this work is to establish a rapid, reliable and reproducible method for visualization of the human corneal SBP using femtosecond laser cut corneal tissue sections. Trephined healthy corneal buttons were fixed and processed using TissueSurgeon-a femtosecond laser based microtome, to obtain thick tissue sections of the corneal epithelium and anterior stroma cut parallel to the ocular surface within approximately 15 min. A near infrared femtosecond laser was focused on to the cornea approximately 70-90 µm from the anterior surface to induce material separation using TissueSurgeon. The obtained corneal sections were stained following standard immunohistochemical procedures with anti-neuronal ß-III tubulin antibody for visualization of the corneal nerves. Sections that contained the epithelium and approximately 20-30 µm of anterior stroma yielded excellent visualisation of the SBP with minimal optical interference from underlying stromal nerves. In conclusion, the results of this study have demonstrated that femtosecond laser cutting of the human cornea offers greater speed, ease and reliability than standard tissue preparation methods for obtaining high quality thick sections of the anterior cornea cut parallel to the ocular surface.

Comparative quantitative assessment of the human corneal sub-basal nerve plexus by in vivo confocal microscopy and histological staining.

PurposeThis study was designed to compare and contrast quantitative data of the human corneal sub-basal nerve plexus (SBP) evaluated by two different methods: in vivo confocal microscopy (IVCM), and immunohistochemical staining of ex vivo donor corneas.MethodsSeven parameters of the SBP in large-scale IVCM mosaicking images from healthy subjects were compared with the identical parameters in ex vivo donor corneas stained by ß-III-tubulin immunohistochemistry. Corneal nerve fiber length (CNFL), corneal nerve fiber density (CNFD), corneal nerve branch density (CNBD), average weighted corneal nerve fiber tortuosity (CNFTo), corneal nerve connection points (CNCP), average corneal nerve single-fiber length (CNSFL), and average weighted corneal nerve fiber thickness (CNFTh) were calculated using a dedicated, published algorithm and compared.ResultsOur experiments showed significantly higher values for CNFL (50.2 vs 21.4 mm/mm2), CNFD (1358.8 vs 277.3 nerve fibers/mm2), CNBD (847.6 vs 163.5 branches/mm2), CNFTo (0.095 vs 0.081 µm-1), and CNCP (49.4 vs 21.6 connections/mm2) in histologically staining specimens compared with IVCM images. In contrast, CNSFL values were higher in IVCM images than in histological specimens (32.1 vs 74.1 µm). No significant difference was observed in CNFTh (2.22 vs 2.20 µm) between the two groups.ConclusionsThe results of this study have shown that IVCM has an inherently lower resolution compared with ex vivo immunohistochemical staining of the corneal SBP and that this limitation leads to a systematic underestimation of several SBP parameters. Despite this shortcoming, IVCM is a vital clinical tool for in vivo characterization, quantitative clinical imaging, and evaluation of the human corneal SBP.

[Magnetic Resonance Microscopy of the Accommodative Apparatus]. / Magnetresonanzmikroskopie des Akkommodationsapparats.

Magnetic resonance microscopy (MRM) at ultra-high magnetic fields allows acquisition of high resolution MR images in the micrometre range. The use of ultra-high magnetic fields opens the possibility of user-independent and artefact-free detailed characterisation of the anatomical tissue of the human eye, which is not achievable with classical imaging techniques. This article correlates MRM of the anterior eye segment and the accommodative apparatus at 9.4 Tesla with conventional histology.

[Finite Element Modelling of the Eye for the Investigation of Accommodation]. / Finite-Elemente-Modellierung des Auges zur Untersuchung der Akkommodation.

Background: Accommodation research increasingly uses engineering methods. This article presents the use of the finite element method in accommodation research. Material and Methods: Geometry, material data and boundary conditions are prerequisites for the application of the finite element method. Published data on geometry and materials are reviewed. It is shown how boundary conditions are important and how they influence the results. Results: Two dimensional and three dimensional models of the anterior chamber of the eye are presented. With simple two dimensional models, it is shown that realistic results for the accommodation amplitude can always be achieved. More complex three dimensional models of the accommodation mechanism - including the ciliary muscle - require further investigations of the material data and of the morphology of the ciliary muscle, if they are to achieve realistic results for accommodation. Discussion and Conclusion: The efficiency and the limitations of the finite element method are especially clear for accommodation. Application of the method requires extensive preparation, including acquisition of geometric and material data and experimental validation. However, a validated model can be used as a basis for parametric studies, by systematically varying material data and geometric dimensions. This allows systematic investigation of how essential input parameters influence the results.

Quantitative assessment of central and limbal epithelium after long-term wear of soft contact lenses and in patients with dry eyes: a pilot study.

PurposeAnalysis of microstructural alterations of corneal and limbal epithelial cells in healthy human corneas and in other ocular conditions.Patients and methodsUnilateral eyes of three groups of subjects include healthy volunteers (G1, n=5), contact lens wearers (G2, n=5), and patients with dry eyes (G3, n=5) were studied. Imaging of basal (BC) and intermediate (IC) epithelial cells from central cornea (CC), corneal limbus (CL) and scleral limbus (SL) was obtained by in vivo confocal microscopy (IVCM). An appropriate image analysis algorithm was used to quantify morphometric parameters including mean cell area, compactness, solidity, major and minor diameter, and maximum boundary distance.ResultsThe morphometric parameters of BC and IC demonstrated no significant differences (P>0.05) between groups. Comparison between three corneal locations (CC, CL, and SL) within the groups showed significant differences (P<0.05) with mean values of cell area, compactness, solidity, and major and minor diameter of BC that increase from CC to limbus. The BC were round and regular in the central cornea (P<0.05) compared with CL and SL.ConclusionsIVCM enables high-quality confocal images from central corneal and limbal epithelium. This quantitative study demonstrated morphological differences in the basal and intermediate epithelium between limbus and central cornea, and found no differences between contact lens wearers, dry eyes, and normal subjects.

[Nonlinear Microscopy in Ophthalmology: Principles and Pathbreaking Applications]. / Nichtlineare Lasermikroskopie in der Ophthalmologie: Physikalische Prinzipien und wegweisende Anwendungen.

Nonlinear microscopy is a non-invasive imaging technique which allows a visualization of biological tissue with high signal contrast due to spectral separation combined with high resolution. In addition to two-photon fluorescence and second harmonic signals also four-wave mixing signals can be used for imaging ocular structures. This review article presents the physical principles of different contrast mechanisms. Exemplary experimental results based on various nonlinear signals are shown, opportunities of this technology are discussed and the prospect of translating this imaging technique into a clinical application is addressed.

[Implant Design by Means of Multiphoton Polymerization]. / Implantatdesign mittels Multiphotonen-Polymerisation.

BACKGROUND: Additive manufacturing and 3D printing create new paths for the design and manufacturing of implants. Technologies with high resolution are required for the development of microstructured eye implants. In the present study, we demonstrate how these technologies can be used during the design development and manufacturing of a multifocal diffractive aspheric intraocular lens. MATERIAL AND METHODS: Multiphoton polymerisation (MPP) is used to manufacture a diffractive relief with resolution in the sub-micrometer range. The relief is applied to the moulded body of a refractive lens, forming a trifocal lens. Propagation of light behind the lens is visualised in water with fluorescein. RESULTS: Multifocal lenses were successfully manufactured with this approach. The optical design with three foci is confirmed by the light propagation images. The images even clearly demonstrate the impact of the refractive and diffractive elements and may provide information on artefacts and aberrations. CONCLUSIONS: Multiphoton polymerisation is an interesting tool for the flexible manufacturing of complex multifocal lenses. With future technological progress in 3D printing with MPP, this is a promising method for on-demand manufacturing of patient individual intraocular lenses.

[Diffusion Weighted Magnetic Resonance Imaging and its Application in Ophthalmology]. / Diffusionsgewichtete Magnetresonanztomografie und ihre potenziellen Anwendungsmöglichkeiten in der Ophthalmologie.

The value of diffusion-weighted magnet resonance imaging (DWI-MRI) has been demonstrated for an ever growing range of clinical indications. DWI is sensitive to the diffusion of water molecules and probes their random displacement within tissue. DWI provides both qualitative and quantitative information on tissue characteristics, e.g. tissue cellularity. This review provides an overview of diffusion-weighted imaging and its emerging applications in ophthalmology. The basic physics and technical foundations of DWI are introduced. The emerging applications of DWI are surveyed, particularly in diseases of the eye, orbit and optical nerve.

[Visual analysis for an early detection of diabetic neuropathy]. / Visuelle Analyse zur Früherkennung einer diabetischen Neuropathie.

Diabetic neuropathy is the most common long-term complication of diabetes mellitus. It comes along with significant nerve dysfunction, which is not reversible. Hence, it is essential to detect nerve fibre abnormalities as early as possible. In this paper, we investigate markers describing degradation of corneal nerves. We apply statistical computations and visual analysis to identify those variables of two clinical studies that separate DN patients from a control group. In this way, the diagnosis of DN patients is supported. The visual analysis is based on different representations visualizing both the statistical results and the gathered multi-variate data. The user can interactively manipulate the views, or select data that will be shown by further displays. In this way, the understanding of the data and its classification is supported. Ambiguous categorisations can be identified and grouped into a so-called "fuzzy group". For this group, further investigations are needed to decide about diabetic neuropathy.

[Large-scale imaging of corneal nerve fibres by guided eye movements]. / Großflächige Abbildung kornealer Nervenfasern durch geführte Augenbewegungen.

BACKGROUND: The high resolution of corneal confocal microscopy (CCM) allows in vivo imaging of the corneal sub-basal nerve plexus (SNP). The field of view of a single CCM image (0.16 mm²) is not sufficient for the reliable morphometric characterisation of the SNP. Therefore we are developing a highly automated mosaicking technique for large-area imaging of the SNP using CCM image sequences. METHODS: In order to acquire an image sequence of a larger area of the SNP, the view direction of the patient is guided by a computer-controlled moving fixation target on a display in front of the non-examined eye. The CCM image sequence is recorded with 30 fps. An online calculated mosaic image allows the medical operator to observe the acquisition process and assess the quality and size of the resulting image during the CCM recording process. Remaining image artefacts are corrected in an automated post-processing step. RESULTS: Using a first prototype system and an appropriate fixation target trajectory, a mean growth of the covered SNP area of 0.18 mm²/s could be achieved. CONCLUSION: Using the presented technology, large-area images of the SNP can be generated. The technology is characterized by a high degree of automation and short examination times.

[Current progress of the artificial accommodation system]. / Fortschritte des Künstlichen Akkommodationssystems.

In case of presbyopia or cataract the "artificial accommodation system" represents one future possibility to durably restore the ability to accommodate. The work presented describes recent progress in the development of the artificial accommodation system. Major advances were achieved in the fields of the actuator system for the active optics, the pupil near reflex sensor, the communication system, the power supply system as well as in system integration. Beside the technical advances, first trials were performed to implant the artificial accommodation system into animals. These trials showed that the new lens shaped design and the C-shaped haptics are beneficial for implantation and secure fixation of the implant inside the capsular bag.

[Optimisation of the visualisation technique for optical paths through intraocular lenses for characterisation of multifocal imaging properties of Fresnel-zone plates]. / Optimierung der Visualisierungstechnik für Strahlenverläufe durch Intraokularlinsen zur Charakterisierung multifokaler Abbildungseigenschaften von Fresnel-Zonenplatten.

The utilisation of the diffractive properties of Fresnel zone plates offers the possibility of intraocular lens designs with multiple foci. Such intraocular lenses can be manufactured by two-photon polymerisation (2PP). This paper explains the underlying concept and shows the principles for visualisation of the focus properties of such implants.

[Ophthalmological imaging with ultrahigh field magnetic resonance tomography: technical innovations and frontier applications]. / Ophthalmologische Bildgebung mit Ultrahochfeld-Magnetresonanztomografie: technische Innovationen und wegweisende Anwendungen.

This review documents technical progress in ophthalmic magnetic resonance imaging (MRI) at ultrahigh fields (UHF, B(0) ≥ 7.0 T). The review surveys frontier applications of UHF-MRI tailored for high spatial resolution in vivo imaging of the eye, orbit and optic nerve. Early examples of clinical ophthalmic UHF-MRI including the assessment of melanoma of the choroid membrane and the characterisation of intraocular masses are demonstrated. A concluding section ventures a glance beyond the horizon and explores research promises along with future directions of ophthalmic UHF-MRI.

[Effect of pseudophacic mini-monovision as an option for independence of spectacles in everyday life]. / Der Effekt der pseudophaken Minimonovision als Option einer Brillenunabhängigkeit im Alltag.

BACKGROUND: Modern day patients who undergo bilateral cataract surgery expect to be spectacles-independent after surgery. The options that are available to attain this goal are pseudophakic monovision, implanting multifocal IOLs or alternatively accommodative IOLs. The aim of this study is to establish the suitability in daily life when attaining minor monovision, named hereafter mini-monovision. MATERIAL/METHODS: In this clinical retrospective study 30 patients were examined. These subjects where bilaterally pseudophake and received either AcrySof®-IOL or Acreos® Adapt-IOL with a slight anisometropia of between 0.5 and 1.75 D and with a maximal astigmatism up to 1.5 D. Visual acuity (VA) for distance (ETDRS), intermediate (Jaeger 80 cm) and near (Jaeger 40 cm) as well as defocus curves were assessed. Furthermore were examined: the reading speed using the font size of the Radner reading chart, anisoconia (FA Oculus according to Esser), stereopsis (Titmus), subjective patient satisfaction (adapted VF-14-questionnaire) and the subjective dependence on having to wear spectacles. RESULTS: The median anisometropia was at 0.75 D; the age was between 61 and 80 (median being at 73 years). 12 patients were male and 18 female. When examined with binocular vision 100 % could attain distance VA of at least decimal 1.0 (LogMAR 0) and an intermediate VA of at least decimal 0.8 (LogMAR 0.1), up to 86.57 % a near VA of at least Jaeger 5, (LogMAR 0.4); up to 63.33 % even managed at least Jaeger 3 (LogMAR 0.3). The median average reading speed for binocular uncorrected reading under photopic conditions was 145 words/minute and under mesopic conditions 117 words/minute. The critical font size was LogRAD 0.60 (Jaeger 5-6), the anisoconia at 2 % and stereopsis at 80 arc seconds. The general dependence on glasses was reduced from 100 % preoperatively to 13 %, postoperatively. Simultaneously a high patient satisfaction score of 93.18 in the VF-14 questionnaire was attained. The result is comparable to those quoted in the literature on full monovision and mulitifocal IOLs. CONCLUSION: To attain spectacles independency, pseudophake mini-monovision is a cost efficient alternative to multifocal lenses for well selected patients. A relative reduction in near VA must be taken into account. To establish the refractive target further examinations are required. Establishing the accurate depth of vision will be helpful in attaining this.

[In vivo MR microscopy of the human eye]. / In-vivo-Magnetresonanzmikroskopie des humanen Auges.

MR microscopy using an ultra high-field MR system is a novel non-invasive imaging technique to explore the human eye without optical distortions. This review aims to provide an insight into the technique. Normal MR microscopic anatomy of the human eye in vivo is demonstrated and clinical applications of MR microscopy are discussed.