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Correlation between optical and psychophysical parameters as a function of defocus.

PURPOSE: To evaluate how ocular optical image quality and psychophysical estimates of visual performance compare to each other as a function of defocus. METHODS: We measured the optical modulation transfer function using a double-pass apparatus and psychophysical estimates of visual performance: contrast sensitivity function (CSF) and visual acuity. Both sets of data were obtained under the same optical conditions. RESULTS: We measured optical and psychophysical parameters as a function of defocus. We studied the correlation between optical parameters (Strehl ratio and the logarithm of the volume in the double-pass image [log_Vol D-P]) and psychophysical parameters (the area under the fitted CSF represented in a logarithmic scale with the spatial frequency in linear scale [Area CSF-log_lin] and visual acuity) for different values of defocus. CONCLUSIONS: Strehl ratio is well correlated with the psychophysical estimates of the visual performance for moderate amount of defocus (within 1 D), whereas the other parameter (log_Vol D-P) is well correlated for larger ranges of defocus (within 2 D) and for different pupil diameters. These results suggest that optical measurements could be used for clinical testing of ophthalmic optics.

Adult↗

Peroral cholangioscopy for the diagnosis and treatment of biliary diseases.

Peroral cholangioscopy with duodenoscopic assistance can allow direct visualization of the bile duct. Several clinical studies suggest the utility of peroral cholangioscopy for the management of various bile duct lesions. Although direct visual observation may be a useful adjunct to endoscopic retro-grade cholangiopancreatography (ERCP) for distinguishing malignant from benign bile duct lesions, the assessment of diagnostic accuracy needs further controlled clinical studies. Intracorporeal lithotripsy with the use of a peroral cholangioscope may be a safe and effective method for difficult-to-treat bile duct stones, including intrahepatic stones. At present, however, the fragility of the fiberscope equipment and technical difficulties hold back its popularity. Preliminary data obtained by using a new videoscope, which provides excellent quality images, are encouraging. Furthermore, it is expected that this videoscope will have longer durability of optical images and better manipulation than previous fiberscopes.

Biliary Tract Diseases↗

Kinetics of intracellular Ca2+ concentration changes and cell contraction of electrically stimulated cardiomyocytes as analysed by automated digital-imaging microscopy.

Enzymatically disaggregated, electrically stimulated cardiomyocytes from adult rats were examined by television-mediated vital microscopy for intracellular Ca2+ concentration and contractile activity. Using an inverted microscope in the epifluorescence mode, the Ca2+ signal was imaged with a low-light-level CCD camera and traced by means of the intracellular concentration of the fluorescent complex of Ca2+ with its indicator Fluo-3. Using the transmitted-light mode, cardiomyocytes that were not loaded were imaged with a conventional CCD camera with automatic gain control and traced by length measurements. Optical images of at least 40 cardiomyocytes per batch of cells from one heart were recorded in up to 20 microscopic fields of observation on videotape within 20 min. They were consecutively analysed by a personal computer installed with an image analysis card at a time-resolution of 20 ms, employing a discrete convolution operation, filtering and threshold setting for fluorescence measurements, and contour description and vectorial analysis for length measurements. Frames of fluorescent images were corrected for the halo effect caused by the increase in the Ca(2+)-dependent fluorescence signal after electrical stimulation. The cell contraction had to be measured in the transmission mode without Fluo-3 due to the inhibition caused by the intracellular Fluo-3. The following coefficients of variation (V) were determined: Vfluorescence < 0.033 and Vtransmission < 0.003 for the precision of measurement, and Vfluorescence < 0.05 and Vtransmission < 0.04 for the reproducibility. The system was validated with isoprenaline and ouabain as agents to modify the Ca(2+)-signal and the contraction. The response of cardiomyocytes of various rats to electrical stimulation, with respect to amplitude and its time point, had a V < 0.08 for both the Ca(2+)-signal and the contraction.

Animals↗

Molecular imaging of factor XIIIa activity in thrombosis using a novel, near-infrared fluorescent contrast agent that covalently links to thrombi.

BACKGROUND: Activated factor XIII (FXIIIa) mediates fibrinolytic resistance and is a hallmark of newly formed thrombi. In vivo imaging of FXIIIa activity could further elucidate the role of this molecule in thrombosis and other biological processes and aid in the clinical detection of acute thrombi. METHODS AND RESULTS: An FXIIIa-sensitive near-infrared fluorescence imaging agent (A15) was engineered by conjugating a near-infrared fluorochrome to a peptide ligand derived from the amino terminus of alpha2-antiplasmin. To evaluate the molecular specificity of A15 for FXIIIa, a control agent (C15) was also synthesized by modifying a single key glutamine residue in A15. Fluorescence imaging experiments with A15 demonstrated stronger thrombosis enhancement in human plasma clots in vitro (P<0.001 versus C15 clots and other controls). A15 was found to be highly specific for the active site of FXIIIa and was covalently bound to fibrin. In vivo murine experiments with A15 demonstrated significant signal enhancement in acute intravascular thrombi (P<0.05 versus C15 group). Minimal A15 enhancement was seen in older aged thrombi (>24 hours), consistent with an expected decline of FXIIIa activity over time. Imaging results were confirmed on correlative histopathology and fluorescence microscopy. CONCLUSIONS: A15 is a novel optical imaging agent that is specifically crosslinked to fibrin by FXIIIa, permitting detection of FXIIIa activity in experimental thrombi in vivo. This agent should permit assessment of FXIIIa activity in a broad range of biological processes and could aid in the clinical diagnosis of acute thrombi.

Animals↗

Molecular imaging of gliomas.

Gliomas are the most common types of brain tumors. Although sophisticated regimens of conventional therapies are being carried out to treat patients with gliomas, the disease invariably leads to death over months or years. Before new and potentially more effective treatment strategies, such as gene- and cell-based therapies, can be effectively implemented in the clinical application, certain prerequisites have to be established. First of all, the exact localization, extent, and metabolic activity of the glioma must be determined to identify the biologically active target tissue for a biological treatment regimen; this is usually performed by imaging the expression of up-regulated endogenous genes coding for glucose or amino acid transporters and cellular hexokinase and thymidine kinase genes, respectively. Second, neuronal function and functional changes within the surrounding brain tissue have to be assessed in order to save this tissue from therapy-induced damage. Third, pathognomonic genetic changes leading to disease have to be explored on the molecular level to serve as specific targets for patient-tailored therapies. Last, a concerted noninvasive analysis of both endogenous and exogenous gene expression in animal models as well as the clinical setting is desirable to effectively translate new treatment strategies from experimental into clinical application. All of these issues can be addressed by multi-modal radionuclide and magnetic resonance imaging techniques and fall into the exciting and fast growing field of molecular and functional imaging. Noninvasive imaging of endogenous gene expression by means of positron emission tomography (PET) may reveal insight into the molecular basis of pathogenesis and metabolic activity of the glioma and the extent of treatment response. When exogenous genes are introduced to serve for a therapeutic function, PET imaging may reveal the assessment of the "location," "magnitude," and "duration" of therapeutic gene expression and its relation to the therapeutic effect. Detailed reviews on molecular imaging have been published from the perspective of radionuclide imaging (Gambhir et al., 2000; Blasberg and Tjuvajev, 2002) as well as magnetic resonance and optical imaging (Weissleder, 2002). The present review focuses on molecular imaging of gliomas with special reference on the status and perspectives of imaging of endogenous and exogenously introduced gene expression in order to develop improved diagnostics and more effective treatment strategies of gliomas and, in that, to eventually improve the grim prognosis of this devastating disease.

Animals↗

Local image structures and optic flow estimation.

Different kinds of local image structures (such as homogeneous, edge-like and junction-like patches) can be distinguished by the intrinsic dimensionality of the local signals. Intrinsic dimensionality makes use of variance from a point and a line in spectral representation of the signal in order to classify it as homogeneous, edge-like or junction-like. The concept of intrinsic dimensionality has been mostly exercised using discrete formulations; however, recent work has introduced a continuous definition. The current study analyzes the distribution of local patches in natural images according to this continuous understanding of intrinsic dimensionality. This distribution reveals specific patterns than can be also associated to local image structures established in computer vision and which can be related to orientation and optic flow features. In particular, we link quantitative and qualitative properties of optic-flow error estimates to these patterns. In this way, we also introduce a new tool for better analysis of optic flow algorithms.

Algorithms↗

[The Array multifocal lens--functional principle and clinical results].

BACKGROUND: Common refractive bifocal IOLs feature 2 or 3 spherical zones with different refraction; the design of the Array multifocal IOL consists of 5 concentric zones, each of them providing a progressive near addition with multiple foci by means of an aspherical surface. METHODS: As a part of a prospective study, distance and near visual acuity, contrast sensitivity, glare and depth of field were examined in 15 patients with the Array lens and a monofocal control group 12 months after implantation. Subjective assessment of the optical quality was performed by detailed asking for optical symptoms. Results were retrospectively compared with the one year follow-up of 15 patients with diffractive bifocal IOLs. RESULTS: No difference was found for best corrected far and near visual acuity and sensitivity for high contrasts. Glare and sensitivity for low contrasts of the monofocal IOL was significantly superior to both multifocal models. Array IOL and diffractive IOL achieved a near acuity of Nieden 2-1 with only distance correction; defocus curves revealed an increase in the depth of field, but with a superiority of the Array lens in intermediate imaging. Optical symptoms as halos were much more frequent in patients with the diffractive IOL than in the Array group. CONCLUSIONS: Functional results of the Array multifocal IOL seem to be comparable to those of the diffractive bifocal IOL, but with the advantage of an improved intermediate imaging and a reduction of optical side effects.

Aged↗

Compact microlenslet-array-based magnifier.

An ultracompact optical imaging system allowing various magnifications or demagnifications and based on microlenslet arrays is presented for the first time to our knowledge. This research generalizes recent findings regarding microlenslet-array-based 1:1 relay systems [Appl. Opt. 42, 6838 (2003)]. Through optical ray tracing, the feasibility of magnifying gray-scale images through a stack of two dissimilar microlenslet arrays is demonstrated for the first time to our knowledge. Results presented specifically demonstrate that a compact imaging system operating at a magnification of 2 is feasible with an overall length of approximately 9 mm. Optical aberrations of the most basic configuration are evaluated, and optimization is discussed.

Journal Article↗

Synaptic physiology: illuminating the road ahead.

A number of genetically encoded reporters of neuronal activity are being developed to assay synaptic activity with single synapse resolution. A recently engineered probe allows imaging of glutamatergic transmission with high sensitivity, and similar probes may help pave the way for optical imaging of excitatory synaptic function in vivo.

Animals↗

Time-resolved optical mammography between 637 and 985 nm: clinical study on the detection and identification of breast lesions.

The first time-resolved optical mammograph operating beyond 900 nm was tested in a retrospective clinical study involving 194 patients with malignant and benign lesions, to investigate the diagnostic potential for the detection and characterization of breast lesions. For the first part of the study (101 patients with 114 lesions), the system was operated at 683, 785, 913 and 975 nm. Subsequently, to improve the spectral content of optical images, the number of wavelengths was increased (up to 7) and the spectral range was extended (637-985 nm). Late gated intensity and scattering images provide sensitivity to tissue composition (oxy- and deoxyhaemoglobin, water and lipids) and physiology (total haemoglobin content and oxygen saturation), as well as to structural changes. Tumours are typically identified because of the strong blood absorption at short wavelengths (637-685 nm), while cysts are characterized by low scattering, leading to a detection rate of approximately 80% for both lesion types, when detection is required in both cranio-caudal and oblique views. The detection rate for other benign lesions, such as fibroadenomas, is presently much lower (<40%). The effectiveness of the technique in localizing and identifying different lesion types was analysed as a function of various parameters (lesion size, compressed breast thickness, age, body mass index, breast parenchymal pattern). The possibility that physiologic changes due to the development of a malignant lesion could affect the entire breast was investigated. The capacity to assess the density of breast based on the average scattering properties was also tested.

Breast Cyst↗

Fluorescence-enhanced optical tomography of a large tissue phantom using point illumination geometries.

We demonstrate fluorescence-enhanced optical imaging of single and multiple fluorescent targets within a large (approximately 1081 cm3) phantom using frequency-domain photon migration measurements of fluorescence collected at individual points in response to illumination of excitation light at individual points on the boundary. The tissue phantom was filled with a 1% lipid solution with and without 0.01 microM Indocyanine Green (ICG) and targets consisted of vials filled with the 1% lipid containing 1-2.5 microM ICG. Measurements were acquired using a modulated intensified CCD imaging system under different experimental conditions. For 3-D image reconstruction, the gradient-based penalty modified barrier function (PMBF) method with simple bounds constrained truncated Newton with trust region method (CONTN) was used. Targets of 0.5, 0.6, and 1.0 cm3 at depths of 1.4-2.8 cm from the phantom surface were tomographically reconstructed. This work demonstrates the practicality of fluorescence-enhanced tomography in clinically relevant volumes.

Breast Neoplasms↗

Cerebellar fastigial nuclei activity during blood pressure challenges.

The cerebellar fastigial nuclei (FN) assist in regulating compensatory responses to large blood pressure changes and show structural injury and functional impairment to cardiovascular challenges in syndromes with sleep-disordered breathing. The patterned time course of FN responses to elevation or lowering of blood pressure and location of responsive regions within the nuclei are unclear. We evaluated FN neural activity in six anesthetized rats using optical imaging procedures during elevation and lowering of arterial pressure by phenylephrine and nitroprusside, respectively. Hypertension diminished optical correlates of FN neural activity, while measures of activity increased to hypotension, with peak neural responses occurring 5-10 s later than peak blood pressure changes. Blood pressure responses were followed by heart rate changes, and peak respiratory rates developed even later, in close temporal proximity to FN activity patterns. Although overall topographical response trends were similar, regional patterns of altered neural activity appeared to both hypertension and hypotension. The extent of neural change was greater during recovery from hypertension than for hypotension at high-dose levels. Blood pressure levels saturated with increasing phenylephrine doses, while FN activity continued to decline. No saturation appeared in heart or respiratory rate trends. The findings suggest that the FN compensate for large blood pressure changes by sympathoexcitatory and inhibitory processes, which accompany late-developing somatic or respiratory adjustments.

Animals↗

Next-generation optical technologies for illuminating genetically targeted brain circuits.

Emerging technologies from optics, genetics, and bioengineering are being combined for studies of intact neural circuits. The rapid progression of such interdisciplinary "optogenetic" approaches has expanded capabilities for optical imaging and genetic targeting of specific cell types. Here we explore key recent advances that unite optical and genetic approaches, focusing on promising techniques that either allow novel studies of neural dynamics and behavior or provide fresh perspectives on classic model systems.

Animals↗

Optical coherence tomography.

The optical coherence tomograph is a new, noninvasive technical device that can obtain cross-sectional, high-resolution images-optical coherence tomographs (OCT)-of the retina. This instrument permits an accurate evaluation of various macular and chorioretinal pathologies and the early detection of glaucomatous damage. Images of the retina are obtained similar to ultrasound B-scan, with 10-microm longitudinal resolution. Because the OCT operates with a near-infrared wavelength (about 840 nm), the examination is of minimal discomfort for the patient. OCT examination is indicated in cases of macular pathologies such as vitreoretinal interface syndrome, in the early detection and quantitative assessment of macular edema, and in the evaluation of a glaucomatous damage by measuring the retinal nerve fiber layer thickness. The future role of this instrument and its applications for clinical diagnosis depend on the future improvement and updating of the software.

Humans↗

Basal ganglia and brainstem encephalitis, optic neuritis, and radiculomyelitis in Epstein-Barr virus infection.

The neurologic complications of Epstein-Barr virus (EBV) infection are rare. We describe a healthy adult with acute EBV meningoencephalomyeloradiculitis. The clinical manifestations, a serologic study, and a dynamic change of EBV DNA in the cerebrospinal fluid with spontaneous recovery confirmed the diagnosis of EBV infection of the nervous system. In addition, we provide other clinical clues for suspicion of EBV infection in patients with encephalitis. These include bilateral basal ganglia and brainstem lesions on magnetic resonance imaging, optic neuritis, or involvement of all levels of the nervous system.

Adult↗

Scanning near-field optical microscopy in cell biology and microbiology.

Scanning a point light source in close proximity over a sample and recording the scattered or transmitted light intensity point by point allows one to record optical images with a resolution not limited by diffraction. An overview of this technique called scanning near-field optical microscopy (SNOM or NSOM) is given with emphasis on cell- and microbiology. After an introduction, where the basic features of the technique are explained, illustrative examples are presented, such as a HeLa cell, fluorescence labelled human chromosomes, super resolution fluorescence imaging, single molecule imaging and fluorescence resonance energy transfer between a single pair of dye molecules.

Chromosomes, Human↗

Detection and staging of esophageal cancers.

PURPOSE OF REVIEW: Technology for detection and staging of esophageal cancer has made significant strides advances in the past 2 years. These advances have led to the enhanced selection of appropriate treatments for esophageal cancer. Cancers that are discovered at an early stage can be treated with endoscopic therapy, whereas advanced cancers are primarily treated with chemotherapy and radiation. RECENT FINDINGS: Detection of esophageal cancer can be enhanced by two major mechanisms: one is by enhancing the lesion, which has typically been done using vital dyes and the other is by changing the method of imaging of the lesion, which has been accomplished by the use of several technologies including fluorescence and optical coherence tomography. Neither of these techniques has been proven, but some investigators have been able to use them to enhance cancer detection. Similar technologies have been applied to staging esophageal cancer. The optical imaging devices also have the potential to stage mucosa-based malignancy. The use of positron emission tomography has been the most recent development that may have application for advanced cancer. Endoscopic ultrasonography has also been improved in resolution and ability to perform fine needle aspiration. The most significant development for staging early cancers is mucosal resection. Finally, by using mucosal resection techniques, the depth of tumor invasion can be established by histology, which allows gastroenterologists to treat early cancers with greater confidence regarding rates of metastatic disease. SUMMARY: Early detection of esophageal cancer can be enhanced by the use of vital dyes for mucosal staining, but the advancement of novel optical diagnostic strategies may be more suitable for clinical use. The primary advantage of these new staging methods is to clearly identify early stage cancer that potentially can be treated without traditional surgical resection techniques. More advanced cancers can be staged with positron emission tomography, but definitive studies demonstrating its role are still lacking.

Journal Article↗

Measurement of human hair growth by optical microscopy and image analysis.

We have developed a quantitative method for measuring hair growth using optical microscopy and image analysis, and have used this to investigate the rate of growth in subjects with and without alopecia. The hairs were cut from an area 7-8 mm in diameter and 24 h and 72 h later, images of the areas were obtained using an optical microscope and were recorded on a video disc. Measurements of the regrowing hairs, placed parallel to the scalp using a glass slide attached to the front of the microscope, were made using the image analyser. In subjects with little or no baldness there was a clear difference between fast-growing hairs and resting or slow-growing hairs. However, in subjects with alopecia there was no such difference and the growth rate of all the hairs showed a continuous distribution. Using this method other parameters such as the number of hairs per unit area and hair diameter as well as grouping of the hairs could be measured.

Adult↗