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[Optical quality after refractive corneal surgery].

Correction of myopia, hyperopia and astigmatism within its indicated margin by means of refractive corneal surgical procedures such as LASIK and surface ablation (e.g. PRK) is one of the standard procedures in ophthalmology. Now that advances in the fields of surgical techniques and the technical devices employed have further progressed in terms of safety and predictability, research also focuses on optical quality. "Optical quality" is not a clearly defined parameter, but can be captured indirectly by means of directly measured data. One has to start with the anatomical properties of the eye, which determine the optical images on the retinal level. The quality of the retinal image influences the eye's function, i.e. acuity and contrast perception. Finally, there is the subjective perception of the image we receive. "Optical quality" as such is reflected by the patient's evaluation of this image perception. Three phenomena are especially responsible for deterioration of the quality of the retinal image: diffraction, aberrations and dispersion. Some of the methods for measuring optical quality are subjective questionnaires, functional testing procedures for measuring visual acuity and contrast sensitivity, optical measuring procedures for the determination of optical quality, as well as biomicroscopy, aberrometry and corneal topography for assessing anatomical changes.

Cornea↗

MR and optical approaches to molecular imaging.

With an increasing understanding of the molecular basis of disease, various new imaging targets have recently been defined that potentially allow for an early, sensitive, and specific diagnosis of disease or monitoring of treatment response. Different approaches to depict these molecular structures in vivo are currently being explored by the molecular imaging community. We briefly review methodologies for molecular imaging by magnetic resonance imaging and optical methods. Special emphasis is put on different contrast agent designs (e.g., targeted and smart probes). New technical developments in optical imaging are briefly discussed. In addition, current research results are put into a clinical perspective to elucidate the potential merits one might expect from this new research field.

Capillary Permeability↗

Evaluation of a device for indirect assessment of automatic exposure control systems.

A test device which can indirectly measure the performance of an automatic exposure control system and which offers the potential to reduce the time and film requirements for such tests has been evaluated. Evaluation results demonstrate that the device can successfully predict image optical density to within +/- 0.10 optical density units over a wide range of imaging technical factors and image optical densities.

Biophysical Phenomena↗

Mapping of stimulus energy in primary visual cortex.

A recent optical imaging study of primary visual cortex (V1) by Basole, White, and Fitzpatrick demonstrated that maps of preferred orientation depend on the choice of stimuli used to measure them. These authors measured population responses expressed as a function of the optimal orientation of long drifting bars. They then varied bar length, direction, and speed and found that stimuli of a same orientation can elicit different population responses and stimuli with different orientation can elicit similar population responses. We asked whether these results can be explained from known properties of V1 receptive fields. We implemented an "energy model" where a receptive field integrates stimulus energy over a region of three-dimensional frequency space. The population of receptive fields defines a volume of visibility, which covers all orientations and a plausible range of spatial and temporal frequencies. This energy model correctly predicts the population response to bars of different length, direction, and speed and explains the observations made with optical imaging. The model also readily explains a related phenomenon, the appearance of motion streaks for fast-moving dots. We conclude that the energy model can be applied to activation maps of V1 and predicts phenomena that may otherwise appear to be surprising. These results indicate that maps obtained with optical imaging reflect the layout of neurons selective for stimulus energy, not for isolated stimulus features such as orientation, direction, and speed.

Animals↗

The use of optical fiber bundles combined with electrochemistry for chemical imaging.

The present Review describes the progress made in using imaging optical fiber bundles for fluorescence and electrochemical-initiated chemiluminescence imaging. A novel optoelectrochemical micro-ring array has been fabricated and demonstrated for concurrent electrochemical and optical measurements. The device comprises optical fibers coated with gold via electroless gold deposition and assembled in a random array format. The design yielded an array of approximately 200 micro-ring electrodes, where interdiffusional problems were minimized. The inner diameter of the ring electrode is fixed by the diameter of the individual optical fibers (25 microns), while the outer radius is determined by the thickness of the deposited gold. While all the fibers are optically addressable, they are not all electrochemically addressable. The resolution of this device is in the tens of micrometers range, determined by the diameter of the optical fiber (25 microns) and by the spacing between each electrically connected fiber. For the purpose of having well-behaved microelectrode characteristics, this spacing was designed to be larger than 60 microns. The array was characterized using ferrocyanide in aqueous solution as a model electroactive species to demonstrate that this microelectrode array format exhibits steady-state currents at short response times. This device has potential application to be used as an optoelectronic sensor, especially for the electrolytic generation and transmission of electrochemiluminescence, and was used to demonstrate that electrochemically generated luminescent products can be detected with the fiber assembly.

Diagnostic Imaging↗

Generation and propagation of subthreshold waves in a network of inferior olivary neurons.

The cells of the inferior olivary (IO) nucleus generate a large repertoire of electrical signals, among them subthreshold oscillations of the membrane potential (STO). To date, subthreshold oscillations have been studied at the level of single-cell recordings, from which network properties were inferred. In this study we used whole cell patch recordings and optical imaging to address the following issues: 1) synchrony of STO in neighboring neurons; 2) stability of the oscillatory activity in the temporal and spatial domain; and 3) the size of the oscillating network. Recordings were made from 126 pairs of IO neurons in 13- to 30-day-old rats. An additional 262 neurons were recorded individually. The frequency of STO varied from 0.8 to 8.6 Hz. The frequency distribution revealed two subpopulations with peaks at about 3 and 6 Hz. The maximum amplitude among the cells varied from 2 to 25 mV. Oscillations in most neurons showed ongoing modulations in both frequency and amplitude. These modulations were largely abolished following bath application of 40 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), a competitive non-N-methyl-D-aspartate (non-NMDA) receptor antagonist, suggesting that they were caused by glutamatergic action. In 35 of 61 recorded pairs at least one neuron exhibited STO permitting us to compare frequency and phase relations. In 22 pairs there was coherent activity with zero phase difference between oscillations in the 2 cells. In these pairs, frequency and amplitude modulation occurred simultaneously in both neurons. Electrotonic coupling was tested in 13 pairs, that had coherent STO, and it was detected in 12. An additional seven pairs showed coherent oscillations but with a phase difference of 20-50 ms. Electrotonic coupling was observed in three of these pairs. Electrotonic coupling was also observed in two of five pairs in which only one neuron oscillated. No coupling was detected in one pair where both neurons oscillated but at different frequencies. Optical imaging using a voltage-sensitive dye (RH 414) was performed on 40 IO slices using an array of 128 photodiodes. Patches of oscillatory activity were observed in 10 slices. Among them six showed spontaneous oscillations, and four exhibited oscillations following extracellular stimulation. In agreement with cell pair recording, optical imaging demonstrated phase-shifted activity in the form of propagating waves of activity within an oscillating patch. We conclude that 1) STO exhibit ongoing modulations of frequency and amplitude that are probably caused by extrinsic inputs to the IO nucleus; 2) electrotonically coupled neurons show a high level of STO synchrony; and 3) the oscillatory activity can propagate within a network of coupled olivary neurons.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

Molecular imaging using visible light to reveal biological changes in the brain.

Advances in imaging have enabled the study of cellular and molecular processes in the context of the living body that include cell migration patterns, location and extent of gene expression, degree of protein-protein interaction, and levels of enzyme activity. These tools, which operate over a range of scales, resolutions, and sensitivities, have opened up broad new areas of investigation where the influence of organ systems and functional circulation is intact. There are a myriad of imaging modalities available, each with its own advantages and disadvantages, depending on the specific application. Among these modalities, optical imaging techniques, including in vivo bioluminescence imaging and fluorescence imaging, use visible light to interrogate biology in the living body. Optimal imaging with these modalities require that the appropriate marker be used to tag the process of interest to make it uniquely visible using a particular imaging technology. For each optical modality, there are various labels to choose from that range from dyes that permit tissue contrast and dyes that can be activated by enzymatic activity, to gene-encoding proteins with optical signatures that can be engineered into specific biological processes. This article provides and overview of optical imaging technologies and commonly used labels, focusing on bioluminescence and fluorescence, and describes several examples of how these tools are applied to biological questions relating to the central nervous system.

Animals↗

Septo-optic dysplasia: MR imaging.

Septo-optic dysplasia is the diagnosis when optic nerve hypoplasia is seen in conjunction with dysgenesis of the septum pellucidum. Nearly two-thirds of these patients have hypothalamic-pituitary dysfunction, and half have schizencephaly. The disorder is difficult to classify because of the diversity of clinical and pathologic manifestations. Magnetic resonance images of 11 patients with clinical and radiographic evidence of septo-optic dysplasia were reviewed retrospectively. The "syndrome" appears to include two subsets of patients whose abnormalities have different embryogenesis and neuropathologic findings. The existence of these two subsets helps to explain the diversity of the clinical and radiologic findings.

Adolescent↗

A new imaging technique for measuring the surface strains applied to dentine.

OBJECTIVES: To investigate possible variation in directional material properties of dentine in relationship to tubule orientation using a new optical imaging technique. METHOD: The optical imaging technique records photometrically a grid pattern formed by using a transmission electron microscope grid as a template on the polished surface of the dentine. The grid pattern is silhouetted onto the sample surface using standard techniques. Compression (c) and diametral compression (dc) tests were undertaken using a servo hydraulic testing machine (MTS model 810) acting on rectangular blocks of dentine with dimensions 1.5 x 1.0 x 1.0 mm (for c) and cylindrical samples with dimensions 2.1 mm in diameter and 1-1.5 mm thick (for dc), respectively. The samples were cut using a diamond wheel and miniature lathe and the cut surfaces polished. Images due to a changing load profile were captured and stored as digitised files on a computer for later analysis. The precision is mainly determined by the pixel resolution of the charged-coupled device camera. RESULTS: Preliminary results show the value of elastic modulus of dentine (10.4 +/- 2.9 GPa) to be similar to those previously reported in the literature. Very small localised strains at the surface of a sample can be observed qualitatively and measured quantitatively by reference to the line spacing (approximately 85 microns). Maximum strength varied with tubule orientation and (compressive/tensile) stress. CONCLUSION: Very small samples of dentine may be investigated for strain in more than one direction using the imaging technique described. These results may be more appropriate for finding relative directional change rather than obtaining the elastic properties of the dentine.

Compressive Strength↗

Noninvasive imaging by optical coherence tomography to monitor retinal degeneration in the mouse.

PURPOSE: Optical coherence tomography (OCT) is a high-resolution imaging technique that measures the intensity of backscattered light from biological microstructures in living tissue. The objective was to evaluate OCT as a routine, noninvasive technique for quantitative measurements of retinal thickness and detachment in small animal models of retinal degenerative diseases. METHODS: An OCT scanning unit was designed and built to visualize retinal tissue from rodents at high resolution in vivo. Several normal and retinal degeneration (rd) mouse strains with different pigmentation, as well as a transgenic mouse strain that carries a wild-type beta-PDE gene in an rd/rd background, were analyzed at different ages. Retinal detachment was induced by subretinal injection of saline. Retinal function was evaluated by full-field ERG, and then each retina was cross-sectionally scanned by OCT. OCT image analysis and measurements of retinal thickness were performed. Animals were then killed and retinal histology was documented. RESULTS: OCT images of the mouse retina revealed structural landmarks allowing assignment of retinal structures. There was no difference in the OCT pattern between pigmented and nonpigmented mice. Changes in the retinal thickness measured by OCT correlated very well with the loss in function measured by ERG and histology in rd/rd and rd/rd/tg(+) transgenic mice at a variety of ages. In addition, retinal detachment caused by surgery was easily visualized and observed by OCT imaging. CONCLUSIONS: OCT imaging is applicable to the mouse retina. There is excellent agreement between the retinal thickness measured by OCT, ERG amplitude, and retinal histology, thus validating OCT imaging as a sensitive and noninvasive tool for monitoring the structural progression of retinal diseases in rodent models. OCT also appears useful for visualizing retinal detachments in the mouse.

Animals↗

Non-invasive imaging of optical parameters of biological tissues.

The normalised back-scattered intensity (NBI) profiles at various locations on the forearms of ten human subjects were obtained by moving the multi-probe of a laser reflectometer. The statistical analysis of the NBI data showed that the variation in the NBI was significantly higher at the ulnar region compared with that at other regions. For determination of the scattering (mus) and absorption (mus) coefficients and the anisotropy parameter g at each location on the forearm, these profiles were matched with the NBI profiles simulated by a Monte Carlo procedure (chi(2)0.99). For the reconstruction of images of variation of these parameters, the averaged values of mua, mus, and g at all locations on the forearms of the subjects were determined. The absorption coefficient had a minimum (1.92 cm(-1)) and maximum (2.21cm(-1)) at the wrist and the lateral region of the forearm, respectively. The scattering coefficient had a maximum (194 cm(-1)) at the medial side and near the elbow, and a minimum (186 cm(-1)) at the lateral side of the forearm. Similar changes in the anisotropy parameter were also observed. By interpolation of the data of each parameter on a 100 x 100 image matrix and after median filtering, colour-coded images of the variation in the optical parameters were constructed. These images could be useful for diagnostic and therapeutic applications of lasers.

Adult↗

Imaging of optically thick specimen using two-photon excitation microscopy.

The in-depth imaging properties of two-photon excitation microscopy were investigated and compared with those of confocal microscopy. Confocal imaging enabled the recording of images from dental biofilm down to a depth of 40 microm, while two-photon excitation images could be recorded at depths greater than 100 microm. Two-photon excitation point spread functions (PSFs) were recorded at depths ranging from 0 to 90 microm depth using 220-nm diameter fluorescent beads immersed in water. PSFs were measured using both a high numerical aperture oil immersion objective and a water immersion objective. The experiments carried out using the oil immersion objective showed a rapid degradation of both the axial and lateral resolution due to spherical aberrations. In addition, the detected fluorescence intensity rapidly decreased as a function of depth. The experiments carried out using the water immersion objective showed no significant degradation of both the axial and lateral resolution and the fluorescence intensity.

Bacteria↗

MR imaging of optic pathways in patients with neurofibromatosis.

Twenty-one patients with documented neurofibromatosis had MR examinations to evaluate possible intracranial disease. In five cases the indication was a known or suspected optic glioma. Two patients were examined because of a history of seizures; the rest were examined as part of a baseline evaluation. Eighteen patients showed evidence of signal hyperintensity on T2-weighted images. Lesions involved the optic nerves, optic chiasm, optic tracts, lateral geniculate body, optic radiations, basal ganglia, periventricular white matter, cerebellar white matter, and dentate nucleus of the cerebellum. Comparison between MR and concurrent CT scans showed MR to be superior in demonstrating the posterior extent of optic-pathway gliomas. In addition, MR showed focal areas of hyperintensity in the basal ganglia, internal capsule, cerebellum, and/or white matter that were not detected on CT. Although we found MR to be superior to CT in detecting intracranial tumors in patients with neurofibromatosis, and in evaluating the extensive involvement of known lesions, the full clinical implications of our findings remain to be determined.

Adolescent↗

Digital imaging of the optic nerve head: monoscopic and stereoscopic analysis.

AIMS: To compare monoscopic and stereoscopic assessment of the optic disc using novel software for the digital stereoscopic analysis of optic disc stereopairs. METHODS: Software was developed for the stereoscopic display of digital optic disc images using an interlaced display method. Neuroretinal rim width was determined at 10 degree intervals around the optic disc using a custom (stereoscopic) cursor whose depth was adjusted to that of Elschnig's rim. Measurements were taken, first viewing the disc monoscopically and at a separate sitting, stereoscopically. RESULTS: Measurements were made in 35 eyes from 35 patients (1260 estimates for each observer) using three observers. The mean cup to disc ratio (CDR) ranged from 0.57 to 0.66 (SD 0.13-0.14) for monoscopic viewing compared with 0.64 to 0.69 (SD 0.12-0.14) for stereoscopic viewing. Stereoscopic assessments gave higher CDRs in temporal, superior, nasal, and inferior aspects of the optic disc (p<0.001, Mann-Whitney U test). Agreement between observers in estimating CDR was high for monoscopic assessment (intraclass correlation coefficient 0.74 (CI 0.72 to 0.76) increasing to 0.80 (0.78 to 0.82) for stereoscopic assessment. CONCLUSION: Digital stereoscopic optic disc assessment provides lower estimates of neuroretinal rim width and higher levels of interobserver agreement compared with monoscopic assessments.

Adult↗

Imaging of optical disc using reflection-mode scattering-type scanning near-field optical microscopy.

A phase-change optical disc was observed using a reflection-mode scattering-type scanning near-field optical microscope (RS-SNOM). In an a.c.-mode SNOM image, the 1.2 microm x 0.6 microm recording marks were successfully observed although the data were recorded on the groove. In contrast, no recording marks could be resolved in a d.c.-mode SNOM image. These results are in good agreement with those from a numerical simulation using the finite difference time domain method. The resolution was better than 100 nm with a.c.-mode SNOM operation and the results indicate that recording marks in phase-change optical media can be directly observed with the RS-SNOM.

Journal Article↗

Imaging techniques in retinal research.

In recent years, retinal research has benefited from major advances in optical imaging approaches. Investigations of the structural and functional organization of the vertebrate retina using live preparations have been facilitated by improvements in cell labeling methods, and by microscopy techniques that permit high-resolution of cells in vitro and in vivo. In particular, the generation of transgenic animals with fluorescently labeled retinal cells has permitted real-time visualization of cell generation, migration, differentiation and growth in the developing retina. Neuronal activity can also be examined by optical imaging using activity reporters directed to specific retinal cell types. Optical techniques such as multiphoton microscopy and total internal reflection fluorescence microscopy (TIRFM) have helped unravel the physiological properties and function of retinal cells. Here, we focus on the latest cell labeling methods that have proven highly useful in many aspects of retinal research. We also highlight several examples of how newly developed imaging technology itself has facilitated investigations that have advanced our understanding of retinal circuits and their development.

Animals↗

Optical methods for imaging ionic activities.

Optical fluorescence is characteristic of molecules and their environment, and dyes can be made whose fluorescence is altered by reversible binding to specific ions. By introducing these into the cytosol, fluorescence microscopy can be used to form dynamic images of ionic activities in living cells under experimental manipulation. Optical fluorescence spectra are broad-band, and if specific ion binding alters the wavelength of maximal excitation or emission, quantitative measurements can be made from the ratio of images taken at two different wavelengths, eliminating errors due to spatial variations in dye concentration and optical path-length. This method is analogous to continuum normalisation in X-ray microanalysis, and is implemented using a sensitive video camera and computer processing digitised images. Fluorescent indicators exist for calcium, magnesium, hydrogen, sodium, zinc and chloride ions. Most imaging work has been on calcium, which is important in many cell signalling processes, and several calcium indicators are available with different spectral properties. Spatial resolution is limited to a few micron by out-of-focus blur, but repeated images can be captured with a time resolution as low as 200 msec, and by using dyes with high binding affinity, detection limits can be lower than by X-ray methods. There is a large and fast-growing literature of applications to many plant and animals cell-types.

Animals↗