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In vivo confocal microscopy in different types of posterior polymorphous dystrophy  [cached]
Babu Kalpana,Murthy K
Indian Journal of Ophthalmology , 2007,
Abstract: Posterior polymorphous dystrophy is a rare corneal dystrophy, usually detected by chance. This case series describes the morphologic features in the three different types of posterior polymorphous dystrophy using confocal microscopy.
Confocal microscopy in the diagnosis of melanoma
Apostolovi?-Stojanovi? Milica,Dobrosavljevi?-Vukojevi? Danijela,La?kovi? Vesna,Stojanovi? A.
Archives of Biological Sciences , 2013, DOI: 10.2298/abs1301279s
Abstract: Melanoma is the most deadly form of skin cancer of melanocytic origin. The tumor has a high malignant potential and early metastasis. Prognosis is directly linked to the stage of the disease. Diagnosing thin melanoma at an early stage offers patients their best chance for survival. The crucial innovation in the early recognition of melanoma was the development of in vivo examination of the skin in high-resolution, by confocal microscopy. Confocal microscopy and its modifications provides a “virtual biopsy“, owing to melanosomes and melanin, which are a source of endogenous contrast. Confocal scanning laser microscopy (CSLM) provides visualization of microanatomical structures and cellular detail in real time (pigmented keratinocytes, melanocytes, melanosomes and melanophages) in the epidermis, dermoepidermal junction and superficial dermis at a resolution equivalent to the resolution of conventional microscopes. [Projekat Ministarstva nauke Republike Srbije, br. 41002]
Basic confocal microscopy  [cached]
Manuela Monti
European Journal of Histochemistry , 2012, DOI: 10.4081/ejh.2012.br3
Abstract: This is an eleven chapter’s effort done by a bunch of Authors coordinated by Prof. R.L. Price and W.G. Jerome (who have personally written almost half of the book) that with great skills are revealing us the secrets of confocal microscopy. Considering the significant progresses in different fields of biology, confocal microscopy is extremely important to dynamically see all the different molecules involved in the controlling networks build up by gene expressions in time and space. Necessary prerequisites to accomplish such goals are some fundamental microscopic technologies well and clearly presented in the first chapters....
Clinical applications of corneal confocal microscopy  [cached]
Mitra Tavakoli,Parwez Hossain,Rayaz A Malik
Clinical Ophthalmology , 2008,
Abstract: Mitra Tavakoli1, Parwez Hossain2, Rayaz A Malik11Division of Cardiovascular Medicine, University of Manchester and Manchester Royal Infirmary, Manchester, UK; 2University of Southampton, Southampton Eye Unit, Southampton General Hospital, Southampton, UKAbstract: Corneal confocal microscopy is a novel clinical technique for the study of corneal cellular structure. It provides images which are comparable to in-vitro histochemical techniques delineating corneal epithelium, Bowman’s layer, stroma, Descemet’s membrane and the corneal endothelium. Because, corneal confocal microscopy is a non invasive technique for in vivo imaging of the living cornea it has huge clinical potential to investigate numerous corneal diseases. Thus far it has been used in the detection and management of pathologic and infectious conditions, corneal dystrophies and ecstasies, monitoring contact lens induced corneal changes and for pre and post surgical evaluation (PRK, LASIK and LASEK, flap evaluations and Radial Keratotomy), and penetrating keratoplasty. Most recently it has been used as a surrogate for peripheral nerve damage in a variety of peripheral neuropathies and may have potential in acting as a surrogate marker for endothelial abnormalities.Keywords: corneal confocal microscopy, cornea, infective keratitis, corneal dystrophy, neuropathy
Confocal Microscopy in Biopsy Proven Argyrosis  [PDF]
Melis Palamar,Suzan Guven Yilmaz,Taner Akalin,Sait Egrilmez,Ayse Yagci
Case Reports in Ophthalmological Medicine , 2013, DOI: 10.1155/2013/875989
Abstract: Purpose. To evaluate the confocal microscopy findings of a 46-year-old male with bilateral biopsy proven argyrosis. Materials and Methods. Besides routine ophthalmologic examination, anterior segment photography and confocal microscopy with cornea Rostoch module attached to HRT II (Heidelberg Engineering GmbH, Heidelberg, Germany) were performed. Findings. Squamous metaplastic changes on conjunctival epithelium and intense highly reflective extracellular punctiform deposits in conjunctival substantia propria were detected. Corneal epithelium was normal. Highly reflective punctiform deposits starting from anterior to mid-stroma and increasing through Descemet’s membrane were evident. Corneal endothelium could not be evaluated due to intense stromal deposits. Conclusion. Confocal microscopy not only supports diagnosis in ocular argyrosis, but also demonstrates the intensity of the deposition in these patients. 1. Introduction Prolonged exposure to silver might cause irreversible pigmentation of the skin (argyria) and/or the eyes (argyrosis) [1]. Hands, eyes, and mucous membranes are affected in most of the patients, and discoloration of the ocular surface is the main ocular evidence [1–3]. A direct relationship was shown between the amount of discoloration and the length of time worked [1]. Confocal microscopy provides high-resolution, high-contrast in vivo images and is a powerful tool for studying the ultrastructure of the cell, its molecular components, and their functions. The Rostock cornea module is an option of the Heidelberg Retina Tomograph (HRT II, Version 3.0; Heidelberg Engineering GMBH, Dossenheim, Germany) introduced as an improvement over older confocal microscopes. The module consists of a monochrome laser radiation source which avoids chromatic aberrations and provides extremely sharp images and a high-powered lens that allows the operator to change the confocal plane within the cornea to capture images at different depths without losing sharpness [4]. The aim of this study is to demonstrate the location of conjunctival and corneal silver deposits by confocal microscopy and to evaluate the correlation between conjunctival histopathology and confocal microscopy findings in a case of occupational argyrosis. To the best of our knowledge this is the first biopsy proven argyrosis case to be evaluated by confocal microscopy. 2. Report of a Case A 46-year-old long-standing silver worker who was diagnosed as ocular argyrosis 4 years earlier was evaluated with confocal microscopy. His visual acuity was 10/20 and intraocular pressures were normal
Twin-Photon Confocal Microscopy  [PDF]
D. S. Simon,A. V. Sergienko
Physics , 2010, DOI: 10.1364/OE.18.022147
Abstract: A recently introduced two-channel confocal microscope with correlated detection promises up to 50% improvement in transverse spatial resolution [Simon, Sergienko, Optics Express {\bf 18}, 9765 (2010)] via the use of photon correlations. Here we achieve similar results in a different manner, introducing a triple-confocal correlated microscope which exploits the correlations present in optical parametric amplifiers. It is based on tight focusing of pump radiation onto a thin sample positioned in front of a nonlinear crystal, followed by coincidence detection of signal and idler photons, each focused onto a pinhole. This approach offers further resolution enhancement in confocal microscopy.
QUANTITATIVE CONFOCAL LASER SCANNING MICROSCOPY  [cached]
Merete Krog Raarup,Jens Randel Nyengaard
Image Analysis and Stereology , 2006, DOI: 10.5566/ias.v25.p111-120
Abstract: This paper discusses recent advances in confocal laser scanning microscopy (CLSM) for imaging of 3D structure as well as quantitative characterization of biomolecular interactions and diffusion behaviour by means of one- and two-photon excitation. The use of CLSM for improved stereological length estimation in thick (up to 0.5 mm) tissue is proposed. The techniques of FRET (Fluorescence Resonance Energy Transfer), FLIM (Fluorescence Lifetime Imaging Microscopy), FCS (Fluorescence Correlation Spectroscopy) and FRAP (Fluorescence Recovery After Photobleaching) are introduced and their applicability for quantitative imaging of biomolecular (co-)localization and trafficking in live cells described. The advantage of two-photon versus one-photon excitation in relation to these techniques is discussed.
Observation of posterior corneal vesicles with in vivo confocal microscopy and anterior segment OCT  [cached]
Ryou Watanabe,Toru Nakazawa,Nobuo Fuse
Clinical Ophthalmology , 2010,
Abstract: Ryou Watanabe, Toru Nakazawa, Nobuo FuseDepartment of Ophthalmology, Tohoku University Graduate School of Medicine, Sendai, JapanAbstract: The histopathology of posterior corneal vesicles (PCV) has not yet been revealed. A 15-year-old girl, who was diagnosed by slit-lamp microscopy as PCV, was examined using specular microscopy, in vivo confocal microscopy, and anterior segment OCT (optical coherence tomography). Anterior segment OCT showed that the thickness of both corneas was within normal limits. At the same time, in vivo confocal microscopy revealed endothelial cells in the rounded dark areas, acellular hyporeflective layers on the Descemet’s membrane, and hyperreflective linear lesions. These findings were not reported previously by slit-lamp and specular microscopy. The abnormal findings only existed at the Descemet’s membrane and corneal endothelial layer. Previous reports dealing with posterior polymorphous dystrophy (PPMD) examined using in vivo confocal microscopy reported almost the same findings, suggesting that PCV and PPMD may be the same at the microstructural level.Keywords: cornea, Descemet’s membrane, imaging
Fluorescent ligands for studying neuropeptide receptors by confocal microscopy
Beaudet, A.;Nouel, D.;Stroh, T.;Vandenbulcke, F.;Dal-Farra, C.;Vincent, J.-P.;
Brazilian Journal of Medical and Biological Research , 1998, DOI: 10.1590/S0100-879X1998001100017
Abstract: this paper reviews the use of confocal microscopy as it pertains to the identification of g-protein coupled receptors and the study of their dynamic properties in cell cultures and in mammalian brain following their tagging with specific fluorescent ligands. principles that should guide the choice of suitable ligands and fluorophores are discussed. examples are provided from the work carried out in the authors' laboratory using custom synthetized fluoresceinylated or bodipy-tagged bioactive peptides. the results show that confocal microscopic detection of specifically bound fluorescent ligands permits high resolution appraisal of neuropeptide receptor distribution both in cell culture and in brain sections. within the framework of time course experiments, it also allows for a dynamic assessment of the internalization and subsequent intracellular trafficking of bound fluorescent molecules. thus, it was found that neurotensin, somatostatin and mu- and delta-selective opioid peptides are internalized in a receptor-dependent fashion and according to receptor-specific patterns into their target cells. in the case of neurotensin, this internalization process was found to be clathrin-mediated, to proceed through classical endosomal pathways and, in neurons, to result in a mobilization of newly formed endosomes from neural processes to nerve cell bodies and from the periphery of cell bodies towards the perinuclear zone. these mechanisms are likely to play an important role for ligand inactivation, receptor regulation and perhaps also transmembrane signaling.
Fluorescent ligands for studying neuropeptide receptors by confocal microscopy  [cached]
Beaudet A.,Nouel D.,Stroh T.,Vandenbulcke F.
Brazilian Journal of Medical and Biological Research , 1998,
Abstract: This paper reviews the use of confocal microscopy as it pertains to the identification of G-protein coupled receptors and the study of their dynamic properties in cell cultures and in mammalian brain following their tagging with specific fluorescent ligands. Principles that should guide the choice of suitable ligands and fluorophores are discussed. Examples are provided from the work carried out in the authors' laboratory using custom synthetized fluoresceinylated or BODIPY-tagged bioactive peptides. The results show that confocal microscopic detection of specifically bound fluorescent ligands permits high resolution appraisal of neuropeptide receptor distribution both in cell culture and in brain sections. Within the framework of time course experiments, it also allows for a dynamic assessment of the internalization and subsequent intracellular trafficking of bound fluorescent molecules. Thus, it was found that neurotensin, somatostatin and mu- and delta-selective opioid peptides are internalized in a receptor-dependent fashion and according to receptor-specific patterns into their target cells. In the case of neurotensin, this internalization process was found to be clathrin-mediated, to proceed through classical endosomal pathways and, in neurons, to result in a mobilization of newly formed endosomes from neural processes to nerve cell bodies and from the periphery of cell bodies towards the perinuclear zone. These mechanisms are likely to play an important role for ligand inactivation, receptor regulation and perhaps also transmembrane signaling.
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