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Results for “Confocal microscopy”

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At least 217 records · Page 12Linked to original sources
PubMed2005

Observations of banding patterns (Vogt striae) in keratoconus: a confocal microscopy study.

PURPOSE: To investigate Vogt striae in keratoconus using confocal microscopy. METHODS: The central cornea of 51 eyes of 29 subjects with keratoconus was observed using a slit-lamp biomicroscope, slit-scanning confocal microscope (TOMEY Confoscan 1), and corneal topographer (EyeSys 2000). RESULTS: Alternating dark and light bands were seen in the stromal images of 23 eyes examined. The bands corresponded with the appearance of Vogt striae on slit-lamp biomicroscopy examination. Bands were found most commonly in the posterior stroma. Posterior bands varied in width, ran mainly in a nearly vertical direction, and appeared to run a straight course through individual image frames. Keratocyte nuclei were located in between the bands. Posterior keratocyte density was unaffected by the presence of bands. Nerve fibers appeared to run a straight course through the bands. When present, bands in the anterior stroma showed greater variability in width and direction within a single frame. Bands were only present in the anterior stroma in more severe levels of keratoconus. The difference in banding pattern noted between the anterior and posterior stroma parallels the known collagen fiber arrangement in the anterior and posterior stroma. CONCLUSIONS: The bands apparent on confocal microscopy of the stroma of the keratoconic cornea correspond with Vogt striae on slit-lamp microscopy. It appears that these bands (and hence Vogt striae) represent collagen lamellae under stress. The stress pattern appears to radiate from the center of the cone and is consistent with the direction of striae when viewed with the confocal microscope.

Adolescent↗
PubMedDate not supplied

[In vivo confocal microscopy in the diagnosis of fungal keratitis: case report].

The authors describe a case of fungal keratitis that the in vivo confocal microscopy helped in the diagnosis and follow-up. Confocal microscopy was done in a patient's ulcer that did not improve with several topical medicines. Corneal scrapings were obtained and culture results were without conclusion. We observed hyphae and infectious collections on confocal microscopy. New corneal culture showed Fusarium sp ten days after confocal diagnosis.

Eye Infections, Fungal↗
PubMed2006

In vivo confocal microscopy of failing and functioning filtering blebs: Results and clinical correlations.

PURPOSE: To correlate clinical filtering bleb function with characteristics as detected by in vivo confocal microscopy. METHODS: In a case-matched cross-sectional study, 52 eyes of 48 patients were examined 1 day to 12.8 years after primary trabeculectomy (mean 375 d). The patients were examined clinically and by in vivo confocal microscopy (Rostock Cornea Module/Heidelberg Retina Tomograph II, Heidelberg Engineering, Inc, Heidelberg, Germany). Nine early and 17 late functioning blebs were pair-matched with malfunctioning blebs. Stromal fiber patterns, the number of intraepithelial and stromal cystic spaces, and the amount of cellular infiltrates were evaluated. RESULTS: Four stromal patterns (trabecular, reticular, corrugated, compacted) invisible to slit-lamp biomicroscopy could be distinguished by in vivo confocal microscopy. The trabecular pattern occurred only in functioning blebs, particularly early postoperatively. Intraepithelial cystic spaces were associated with functioning late blebs, whereas they were equally distributed in early blebs. In contrast, stromal cystic spaces indicate function in early blebs, whereas in late blebs the number of these cavities was similar in both groups. The density of intraepithelial and stromal round cells was higher in functioning late blebs compared with malfunctioning late blebs. CONCLUSIONS: In vivo confocal microscopy allows to assess filtering bleb structures that are invisible biomicroscopically. Some morphologic features detected by this technique seem to indicate filtering bleb function and time after surgery. The predictive value of these features deserves further clarification in a prospective longitudinal study.

Aged↗
PubMed2006

Spectrally encoded slit confocal microscopy.

A simple and cost-effective method for real-time imaging in confocal microscopy is proposed. Spectrally encoded slit confocal microscopy (SESCoM) uses a spectral encoding technique together with a confocal slit aperture to achieve two-dimensional images. Simulation and experimental results of the SESCoM's axial and lateral performances are presented. The measured FWHM of the axial response is 1.15 mum when an objective with a NA of 0.95 is used. FWHMs of the lateral line spread functions are measured to be 236 and 244 nm along the x and y directions, respectively. Both the axial and the lateral experimental results agree well with the simulation results.

Equipment Design↗
PubMed1993

Three-dimensional distribution of damaged cells in cryopreserved pancreatic islets as determined by laser scanning confocal microscopy.

The technique of serial optical sectioning by confocal microscopy, in conjunction with off-line digital image analysis, was used to quantify the radial distribution of damaged cells in rat pancreatic islets following cryopreservation. The process consists of imaging frozen-thawed islets of Langerhans using laser scanning confocal microscopy (LSCM). The three-dimensional (3-D) distribution and analysis of the two populations of viable and damaged cells was visualized via acridine orange/propidium iodide (AO/PI) fluorescent staining. In preparation for cryopreservation, isolated and cultured rat pancreatic islets were brought to a 2 M concentration of dimethyl sulphoxide (DMSO) by serial addition at decreasing temperatures. Ice was nucleated in the islet suspension at -10 degrees C, and individual specimens were frozen to -70 degrees C at cooling rates of 1, 3, 10 and 30 degrees C/min in a programmable bulk freezer and subsequently stored in liquid nitrogen. After rapid thawing and serial dilution to remove DMSO, individual islets were prepared with AO/PI stains for imaging on the LSCM. Serial sections of the islets, 2-7 microns in thickness, were obtained and processed to obtain high-contrast images. Analysis algorithms consisted of template masking, grey-level thresholding, median filtering and 3-D blob colouring. The radial distribution of damaged cells in the islets was determined by isolating the cell and computing its distance from the centroid of the 3-D islet volume. An increase in the number of blobs corresponding to single and/or aggregates of damaged cells was observed progressively with distance from the centre towards the periphery of the islet. This pattern of freeze-induced killing of cells within the islet was found to occur consistently in the numerous individual specimens processed.

Animals↗
PubMed2006

In vivo confocal microscopy of normal conjunctiva and conjunctivitis.

PURPOSE: To analyze the appearance of normal conjunctiva and conjunctival inflammation by in vivo confocal microscopy. METHODS: Conjunctiva of 15 normal patients and 21 patients with conjunctivitis including bacterial, papillary, follicular, granulomatous, and cicatrizing disease were analyzed by the Heidelberg retina tomograph (HRTII)/Rostock cornea modul (RCM). RESULTS: Scans of normal bulbar and tarsal conjunctiva corresponded well to the established anatomy except for a prominent, thickened epithelial basement membrane observed by in vivo microscopy. Presumed goblet cells were visible throughout the conjunctival epithelium. Adenoid structures and hair follicles were discernible in the tarsal conjunctiva in vivo. Conjunctival perfusion could be observed directly. Acute and chronic inflammatory cells, conjunctival papillary, and follicular reactions, as well as conjunctival cicatrization, could be discriminated. In a patient with conjunctival granuloma, in vivo confocal microscopy disclosed suture material inside the lesion. CONCLUSION: Confocal microscopy using near-infrared laser light is a useful new tool in the analysis of conjunctival tissue in vivo. It is a valuable aid in the differential diagnosis of conjunctival inflammation and thus may guide therapeutical decisions.

Adult↗
PubMed2002

[Diffuse lamellar keratitis after laser in situ keratomileusis. Clinical and confocal microscopy findings].

BACKGROUND: Diffuse lamellar keratitis (DLK) is a sterile inflammation after lamellar corneal surgery. A clinical staging for DLK was recently set up. In this observational case series, the clinical and confocal microscopic findings of stages 1-3 are reported. METHODS: Six eyes of six patients (three eyes with DLK stage 1, two eyes with DLK stage 2, and one eye with DLK stage 3) were examined by slit lamp biomicroscopy and confocal microscopy. RESULTS: In all cases, confocal microscopy showed an infiltration of inflammatory cells into the anterior stroma and the flap interface. The number of cells varied between the eyes with DLK stage 1, and both stage 2 corneas had dense infiltrates. In the eye with stage 3 DLK, an aggregation of decayed cells, most likely granulocytes, was noticed clinically and by confocal microscopy. CONCLUSIONS: The DLK stages represent different clinical intensities of interface inflammation after LASIK. While stages 1 and 2 have a similar confocal microscopic appearance, stage 3 is a result of aggregation of a high amount of inflammatory cells.

Adult↗
PubMedDate not supplied

Diagnostic efficacy of endometrial cytology with the Abradul cell sampler supplemented by laser scanning confocal microscopy.

OBJECTIVE: To study the diagnostic efficacy of endometrial cytology with the Abradul cell sampler and the effect of supplementing it, in cases with overly thick smears, with laser scanning confocal microscopy. STUDY DESIGN: Sampling was performed in 1,684 women. All patients underwent subsequent histology. In eight cases with overly thick smears the original smears were restained to allow confocal microscopy. RESULTS: Efficacy was high, with 1,593 good samples. Thirty-one endometrial carcinomas were signed out as cytologically positive and 10 as atypia. In addition, 10 squamous cell carcinomas of the cervix and 1 ovarian carcinoma were diagnosed with this method. Only one case (endocervical adenocarcinoma) was not detected in the sample. In the eight cases in which confocal microscopy was performed, the images were high quality, allowing a correct diagnosis. In addition, the transition from hyperplasia to carcinoma could be visualized in the cytologic smear. CONCLUSION: Endometrial cytology in an outpatient setting is highly effective, and confocal microscopy in selected cases is not only helpful but also highly instructive.

Disposable Equipment↗
PubMedDate not supplied

Confocal microscopy of stage 4 diffuse lamellar keratitis with spontaneous resolution.

PURPOSE: To report confocal microscopic findings at the onset of stage 4 diffuse lamellar keratitis and after its resolution. Stage 4 is the most severe form of diffuse lamellar keratitis. Its incidence is approximately 1 in 5000 and is associated with stromal melting, deep flap folds, central haze, hyperopic shift, irregular astigmatism, and severe decrease in visual acuity. METHODS: A 22-year-old woman underwent bilateral uncomplicated laser in situ keratomileusis (LASIK) for myopia. Postoperative course in the right eye was uneventful; however, in the left eye, stage 4 diffuse lamellar keratitis developed. Confocal microscopy examination was performed in both eyes at the onset of the syndrome and after its resolution. Findings in the eye with diffuse lamellar keratitis (left eye) were compared with the uninvolved (right) eye. RESULTS: The condition improved spontaneously and 2 years later, slit-lamp microscopy showed resolution of the folds and haze with subsequent improvement of visual acuity. However, confocal microscopic examination in the left eye revealed a persistent stromal subclinical haze on both sides of the lamellar cut and prominent folds that extended into the anterior stroma. CONCLUSION: Confocal microscopy revealed that in spite of normal appearance on slit-lamp microscopy, micromorphological alterations persisted after spontaneous resolution of stage 4 diffuse lamellar keratitis.

Adult↗
PubMed1996

Corneal reinnervation following penetrating keratoplasty--correlation of esthesiometry and confocal microscopy.

Thus far assessment of corneal reinnervation after penetrating keratoplasty has been possible only by esthesiometry techniques. Since the introduction of confocal microscopy, we have been capable of performing structural corneal in vivo examinations. The purpose of our study was to correlate esthesiometry results with confocal microscopy findings. We used a Cochet-Bonnet esthesiometer and a Microphthal confocal microscope to investigate corneal grafts in vivo. A total of 46 eyes were examined preoperatively and for up to 3 years after penetrating keratoplasty. At 8 weeks after keratoplasty the first stromal nerves were detected in the periphery of the graft. The first nerves in the middle and superficial stroma of the graft center were observed at 7 months after surgery. Reinnervation of the central basal epithelium was found at 2 years after keratoplasty. The highest level of sensitivity was detected in young patients with reinnervation of the basal epithelium. Confocal microscopy enables us to correlate morphologically and functionally corneal reinnervation after surgery. The present study shows that corneal reinnervation is influenced by the amount of time elapsing after surgery, the patient's age, and the preoperative diagnosis. In none of our patients was normal nerve morphology or sensitivity observed during the follow-up period. Comparison of the morphology of nerves seen in eyes after nonsurgical trauma with that observed in corneal grafts indicates that surgically induced scar formation may limit nerve regeneration in grafts.

Adolescent↗
PubMed2006

Fuzzy logic and maximum a posteriori-based image restoration for confocal microscopy.

We propose a maximum a posteriori image restoration approach to 3D confocal microscopy. The image field is suitably modeled as a Markov random field, resulting in a Gibbs distributed image. A fuzzy-logic-based potential is employed in the Gibbs prior. Unlike other potentials, the fuzzy potential distinguishes intensity variation due to genuine edges and noise. The proposed approach has generated artifact-free restored confocal microscopy images.

Algorithms↗
PubMed2004

Quality assessment model of 3 different microkeratomes through confocal microscopy.

PURPOSE: To study the quality of the cut created by 3 microkeratomes from 2 different generations using corneal confocal microscopy. SETTING: Department of Refractive Surgery, Instituto Oftalmológico de Alicante, Alicante, Spain. METHODS: Two different studies were conducted: Study 1 and Study 2. Study 1 was a prospective analysis using confocal microscopy examination data from 2 reference groups: Group A (control) with 20 nonoperated eyes of 20 healthy volunteers, and Group B with 50 eyes of 30 patients operated on with the Bausch & Lomb Automated Corneal Shaper (ACS) microkeratome. Study 2 was a prospective randomized double-masked study in which 40 eyes of 20 patients underwent myopic laser in situ keratomileusis by 1 surgeon. The right and left eyes of each patient were randomly and alternatively assigned to Group C, flap made with the Hansatome microkeratome, or Group D, flap made with the Moria M2. Immediately after surgery, the microscopic appearance of the cut was subjectively evaluated by the surgeon. One month postoperatively, flap thickness, particle density, and the subclinical confocal wound healing opacity (WHO) index were evaluated with the corneal confocal microscope. RESULTS: Surgeon handling comfort was nearly the same with the Hansatome as with the M2 (P =.540). However, the apparent quality of the resulting flap was better with the M2 microkeratome (P =.041). The depth of the cuts made by the 3 microkeratomes were significantly different (P<.001), with the ACS flaps being thinner than the flaps made with the Hansatome or the M2. Particle density at the interface was significantly poorer in the eyes operated on with the ACS, but in these cases the WHO index was significantly greater (P<.001 in both cases). CONCLUSIONS: Confocal microscopy is a very useful tool to evaluate the quality of the cut made by different microkeratomes. Overall, the predictability in flap thickness and the apparent quality of the cut made by the Moria M2 microkeratome are better than those obtained with the Hansatome or the ACS. The thin flap made by the ACS microkeratome produced a significantly greater WHO index than the thicker flaps created with the other 2 microkeratomes.

Corneal Stroma↗
PubMed1997

Fact and artefact in confocal microscopy.

High-resolution confocal microscopic images may be made of either the surface of a sample or beneath the surface. These images can be likened to optical tomograms, giving thin (> 0.35 micron) slices up to 200 microns below the surface of a transparent tissue: With microscopes running under normal conditions, the optical section thickness will be > 1 micron and the effective penetration into enamel and dentin a maximum of 100 microns. For maximum resolution, high-quality, high-numerical-aperture objectives should be used. Refractive index matching of the lens immersion media and the substrate will avoid distortions of images in the optical axis. Such errors could occur when imaging a considerable distance (> 40 microns) into a cell containing water, with an oil immersion objective above the cover slip. Care should be taken in the interpretation of computerized z axis reconstructions made from serial optical sections: Their validity should be checked with equivalent views made with the sample oriented in the same direction as the reconstruction. The use of fluorescent dyes in microscopy is a very powerful investigative technique. It is important that the dyes used not be labile and that they be well-fixed to the materials being examined, or the images may indicate the dye distribution rather than the material to which it is "attached". Multiple labeling experiments must have crossover control experiments to verify the distribution of the individual dyes. Valuable information can often be gained by combining information from both reflection and fluorescence images. Two-photon laser excitation of dyes gives the potential for greater depth penetration and improved resolution.

Artifacts↗
PubMed2004

Confocal microscopy and corneal sensitivity in a patient with corneal manifestations of Tangier disease.

PURPOSE: Tangier disease is an autosomal recessive disorder in which cholesterol-rich lipids are deposited at various tissues of the body including the cornea. In this case report, the corneal changes in a patient with Tangier disease are described. METHODS: A 72-year-old patient who was diagnosed with Tangier disease 25 years before received a complete eye examination including confocal microscopy and Cochet-Bonnet esthesiometry. RESULTS: Slit-lamp biomicroscopy and confocal microscopy showed bilateral corneal opacifications caused by lipid accumulation. Confocal microscopy showed that pathologic changes in the cornea in Tangier disease are limited to the stroma. Neither a reduced corneal sensation nor lid abnormalities as previously described in Tangier disease were found. CONCLUSION: Confocal microscopy helps to identify corneal changes in the stroma caused by Tangier disease easily missed in a slit-lamp examination. The ocular manifestations of Tangier disease do not necessarily include a reduced corneal sensitivity and lid abnormalities.

Aged↗
PubMed2005

[In vivo confocal microscopy and ocular surface diseases: anatomical-clinical correlations].

PURPOSE: To describe corneal and conjunctival epithelial changes in various ocular surface disorders with a new in vivo confocal microscope (Corneal HRT II module) and to compare these results with those obtained by immunohistology on impression cytology specimens. METHODS: In this study, we investigated 36 eyes of 18 patients with ocular surface diseases such as corneal epitheliopathy induced by ocular rosacea (five patients), keratoconjunctivitis sicca related to tear evaporative condition in Meibomian gland dysfunction (eight patients), or tear-deficient dry eye in Sjögren's syndrome (five patients). In vivo confocal microscopy imaging (Heidelberg Retina Tomograph II Rostock Cornea Module) and impression cytology of the cornea and/or conjunctiva were performed. The images were analyzed for epithelial cell morphology at nuclear and cytoplasmic levels, in the cornea and the conjunctiva, number of goblet cells and their possible presence within the corneal epithelium, corneoconjunctival junction at the limbus, and density and presence of inflammatory cells within both epithelia. RESULTS: Images obtained consisted of two-dimensional 400x400 microm optical sections oriented parallel to the surface of the eye and compared with impression cytology. Conjunctival and corneal epithelia showed dramatic changes in ocular surface disease. In keratoconjunctivitis sicca, we found squamous metaplasia, inflammatory cell infiltration, goblet cell depletion, as well as a nuclear snake-like chromatin pattern. In rosacea associated with corneal epitheliopathy, a corneal conjunctivalization (goblet cells together with conjunctival epithelial cells within the corneal epithelium layer) was found. The images provided by this technique are in excellent correlation with the findings using impression cytology. CONCLUSION: The HRT II in vivo confocal microscope facilitates the study of both central and peripheral ocular surface epithelia compared to first-generation confocal microscopy devices. In vivo confocal microscopy, showing corneal and conjunctival epithelial structures in the live eye at the cellular level, is likely to open up a new promising way to investigate ocular surface involvement in complex diseases, providing a new insight on corneal and conjunctival disorders in the future.

Conjunctiva↗
PubMed2004

[Confocal microscopy: a tool to visualise differentiation of stem cells into cardiomyocytes].

Confocal microscopy offers important advantages compared to conventional epifluorescence microscopy. It works as an "optical microtome" leading to a accurate image resolution of a defined focal plane. Furthermore, the addition of a Nipkow disk on the confocal microscope greatly accelerates the image acquisition, up to 30 frames per second. Nevertheless, the software-assisted mathematical restoration of images acquired using a wide-field microscope allows to get images with a resolution similar to the one obtained in confocal microscopy. These imaging technologies allowed us to monitor on line cardiac differentiation of murine embryonic stem (ES) cells within 3D structures called embryoid bodies. The high rate acquisition of images using the confocal microscope equipped with a Nipkow disk allows to monitor calcium spiking in differentiating cardiomyocytes within embryoid bodies.

Animals↗
PubMed2006

An algorithm for 3-D refractive index measurement in holographic confocal microscopy.

The principle of holographic confocal microscopy is introduced in this paper. An algorithm is presented to retrieve the phase information from the holograms and calculate the refractive index of the specimen from the phase. It is shown that this algorithm can be applied to a refractive index distribution in which the variation in x and z directions can be separated. The two examples used for the simulations are a linear gradient distribution and a circular distribution, which can be found in practical situations.

Journal Article↗
PubMedDate not supplied

Mitochondria in tumor cells studied by laser scanning confocal microscopy.

We present here a confocal fluorescence microscopy study of mitochondria in sensitive and resistant carcinoma cells by using two potentiometric probes of mitochondria, rhodamine 123 (R123) and dimethylaminostyryl-methylpyridiniumiodine. We have found that active mitochondria in sensitive MCF-7 and multidrug resistant MCF-7/DX carcinoma cells are very different in localization and morphology. In sensitive cells active mitochondria are found in the perinuclear region, whereas in the multidrug resistance (MDR) subline they are confined to the cell periphery. Interestingly, the MDR revertant verapamil has been found to restore in MCF-7/DX cells the same pattern of active mitochondria seen in sensitive cells. We have also studied R123 in human lung carcinoma A549 cells, which display a low responsivity to doxorubicin, and overexpress the lung resistance-related protein. In addition to perinuclear mitochondria, peripheral mitochondria with weaker fluorescence can be seen in this cell line. Interestingly, in the two examined carcinoma lines we have been able to recognize by image analysis a common new star-lobed morphology. Our results indicate that in resistant carcinoma cells two populations of mitochondria coexist with different localization, morphology, and activity.

Adenocarcinoma↗