Confirmation by magnetic resonance imaging of optic nerve injury after retrobulbar anesthesia.
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Magnetic resonance imaging (MRI) has become an important diagnostic modality for the evaluation of patients with chiasmal or perichiasmal disease. In this review, normal chiasmal anatomy and various pathologic conditions will be illustrated to demonstrate the advantages of MRI over computed tomography (CT). MRI is the neuroimaging method of choice in chiasmal evaluation, but sometimes it may be complemented by CT scanning.
Optical imaging, such as transmission imaging, is used to study brain tissue injury. Transmission imaging detects cellular swelling via an increase in light transmitted by tissue slices due to a decrease in scattering particle concentration. Transmission imaging cannot distinguish sub-cellular particle size changes from cellular swelling or shrinkage. We present an optical imaging method, based on Mie scatter theory, to detect changes in sub-cellular particle size and concentration. The system uses a modified inverted microscope and a 16-bit cooled CCD camera to image tissue light scatter at two angles. Dual-angle scatter ratio imaging successfully discriminated latex microsphere suspensions of differing sizes (0.6, 0.8, 1 and 2 microm) and concentrations. We applied scatter imaging to hippocampal slices treated with 100 microM N-methyl-D-aspartate (NMDA) to model excitotoxic injury or -40 mOsm hypotonic perfusion solution to cause edema injury. We detected light scatter decreases similar to transmission imaging in the CA1 region of the hippocampus for both treatments. Using our system, we could distinguish between NMDA and hypotonic treatments on the basis of statistically significant (P<0.0003) differences in the scatter ratio measured in CA1. Scatter imaging should be useful in studying tissue injuries or activity resulting in brain tissue swelling as well as morphological changes in sub-cellular organelles such as mitochondrial swelling.
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BACKGROUND: Optical coherence tomography (OCT) is a new technology capable of generating high resolution cross-sectional images of complex tissue in real time. Analogous to ultrasound, OCT measures backscattered light intensity using coherence interferometery to construct topographical images of complex tissue. Since OCT uses infrared light rather than acoustic waves, its spatial resolution is exceptionally high (2-10 microm). Recent advances in data acquisition, analysis, and processing enable real-time imaging, and make OCT a potentially valuable tool for pulmonary airway diagnostic applications, including assisting directed airway biopsies. OBJECTIVE: This study evaluates feasibility of OCT for delineating proximal airway microstructures in various animal as well as human tracheas. METHODS: Excised trachea samples from New Zealand white rabbits, Duroc pigs, and human trachea were imaged using a compact, 1,300-nm broad-band superluminescent-diode-based prototype fiber OCT device we constructed. The resulting structural OCT images were compared to conventional hematoxilin and eosin (HE) stained histological sections from the same samples. RESULTS: OCT was able to delineate microstructures such as the epithelium, mucosa, cartilage, and glands in all samples. CONCLUSION: These findings suggest that integration of OCT with flexible fiberoptic bronchoscopy could enhance pulmonary diagnostic medicine and detection of pathologic tissue changes in various respiratory diseases.
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OBJECTIVE: The conventional method for the analysis of myocardial cell transplantation depends on postmortem histology. Here, we have sought to demonstrate the feasibility of a longitudinal monitoring of transplanted cell survival in living animals, accomplished with optical imaging techniques and pharmacological interventions. METHODS: Human cord blood (50 ml) was donated with parental consent. After getting cord blood derived mesenchymal stem cells (CBMSCs), cells were transfected (MOI = 100) overnight with adenovirus encoding firefly luciferase gene (Ad-CMV-Fluc). Our experimental Sprague-Dawley rats (n = 12) were given intramyocardial injections containing 1 x 10(6) CBMSCs, which had been made to express the firefly luciferase (Fluc) reporter gene. Optical bioluminescence imaging was then conducted using a cooled charged-coupled device (CCD) camera (Xenogen), beginning on the day after the transplantation (day 1). Groups of mice were intraperitoneally injected with cyclosporine (5 mg/kg) or tacrolimus (1 mg/kg), in an attempt to determine the degree to which cell survival had been prolonged, and these values were then compared with the cell survival values of the negative control group. The presence of transplanted CBMSCs on in vivo images confirmed by in situ hybridization for human specific Alu in the myocardium. RESULTS: Cardiac bioluminescence signals were determined to be present for 6 days after transplantation: day 1 (97000 +/- 9100 x 10(5) p/s/cm2/sr), day 3 (9600 +/- 1110 p/s/cm2/sr), and day 5 (3200 +/- 550 p/s/cm2/sr). The six mice that received either cyclosporine or tacrolimus displayed cardiac bioluminescence signals for a period of 8 days after transplantation. We observed significant differences between the treated group and the non-treated group, beginning on day 3 (tacrolimus; 26500 +/- 4340 p/s/cm2/sr, cyclosporine; 27200 +/- 3340 p/s/cm2/sr, non-treated; 9630 +/- 1180 p/s/cm2/sr, p < 0.01), and persisting until day 7 (tacrolimus; 12500 +/- 2946 p/s/cm2/sr, cyclosporine; 7310 +/- 1258 p/s/cm2/sr, non-treated; 2460 +/- 160 p/s/cm2/sr, p < 0.01). The human-derived CBMSCs were detected in the myocardium 3 days after transplantation by in situ hybridization. CONCLUSIONS: The locations, magnitude, and survival duration of the CBMSCs were noninvasively monitored with a bioluminescence optical imaging system. We determined that optical molecular imaging expedites the fast throughput screening of pharmaceutical agents, allowing for the noninvasive tracking of cell survival within animals. In rat cardiac CBMSC transplant models, transient immunosuppressive treatment with tacrolimus or cyclosporine was shown to improve donor cell survival.
PURPOSE: To compare the diagnostic performance of the Heidelberg Retinal Tomograph's (HRT; Heidelberg Engineering GmbH, Dossenheim, Germany) glaucoma probability score (GPS), an automated, contour line-independent method of optic disc analysis with that of the Moorfields regression analysis (MRA). METHODS: HRT images were obtained from one eye of 121 patients with glaucoma (median age, 70.2 years; median mean deviation [MD], -3.6 dB, range, +2.0 to -9.9 dB) and 95 healthy control subjects (median age, 59.7 years; median MD -0.1 dB, range +2.5 to -3.7). The diagnostic performances of GPS and MRA were evaluated by including borderline classifications, either as test negatives (most specific criteria) or as test positives (least specific criteria). Agreement between global and sectoral data of both analyses was established. Logistic regression analyses were performed to evaluate the effect of covariates such as optic disc size and age on the classification outcomes of both the GPS and the MRA. RESULTS: In 8 (7%) patients with glaucoma and 10 (11%) control subjects, the GPS failed to provide a complete global and sectoral optic disc classification. Although we could not identify a single distinct cause of this failure in the glaucoma group, failures in the control subjects occurred most often (7/10) with small and crowded optic discs. In subjects who were successfully classified at least globally by the GPS (117 patients with glaucoma, 88 control subjects), the diagnostic performances of GPS and MRA were similar (areas under the receiver operating characteristic [ROC] curve of 0.78 and 0.77, respectively; P > 0.1). With the GPS, sensitivity and specificity were 59% and 91% (most specific criteria) and 78% and 63% (least specific criteria), respectively. Combining GPS and MRA did not increase diagnostic performance significantly (ROC area of combined classifiers, 0.81). Both GPS and MRA were affected by disc size. In patients with glaucoma as well as healthy control subjects, the odds of a positive GPS classification (borderline or outside normal limits) increased by 21% (95% confidence interval [CI], 12%-30%) for each 0.1 mm2 increase in optic disc area. With the MRA, the corresponding increase was 15% (95% CI, 7%-23%). Optic disc area alone accounted for approximately 30% and 22% of the explained variance with the GPS and MRA, respectively (P < 0.001). The proportional-odds logistic regression confirmed that optic disc size affected mainly the tradeoff between true- and false-positive classifications (criterion) rather than the absolute performance of the analyses (area under the ROC curve). There was some evidence of an age effect with the MRA, which showed a 53% (95% CI, 16%-102%) increase in the odds of a positive test (borderline or outside normal limits) associated with each decade of age (P = 0.002), but no age effects were observed with the GPS (P > 0.1). CONCLUSIONS: The diagnostic performance of the contour line-independent GPS analysis is similar to that of the MRA. However, clinicians should be aware of the strong size dependence of both GPS and MRA. In large optic discs, both GPS and MRA are likely to produce many false-positive classifications. Correspondingly, the sensitivity to early damage is likely to be low in small optic discs. There is a need for automated classification systems that explicitly address the size dependence of current analyses.
If an optically active (chiral) sample is placed in a microscope and illuminated with circularly polarized light, an image can be formed that is related to the circular dichroism of each feature of the sample. A theoretical investigation has been done for the circular differential image obtained by subtracting the images formed under right- and left-circularly polarized light. Two types of differential images are possible: (i) dark-field images formed from light reflected or scattered by the sample and (ii) bright-field images formed from light transmitted through the sample. The sign and magnitude of each feature in a circular differential image strongly depend on the structure of the sample. The dark-field circular differential images are most sensitive to large features with dimensions similar to the wavelength of illumination whereas the bright-field images are most sensitive to the short-range molecular order. Applications of circular differential imaging may include clinical fingerprinting of normal and transformed cells and structural analysis of individual cellular components.
We describe a new one-dimensional synthetic aperture imaging laser radar (ladar) using the resonant sensitivity of a microchip laser to frequency-shifted optical feedback and galvanometric scanning of the target under investigation. In our experiment, the laser is both the source and the detector, providing optical amplification with self-aligned heterodyne detection. By using galvanometric scanning, we achieve an along-track spatial resolution better than the diffraction limit.
Acquired pendular nystagmus (APN) complicates multiple sclerosis and other neurological disorders, causes visual impairment, and is often refractory to available treatments. Vision could be improved by an optical aid that shifts the seen world in lockstep with the APN. An essential component of such a device is the image-shifting mechanism, which must be light, accurate, suitable for battery operation, and capable of image shifting at the frequencies and amplitudes seen in APN. We determined that a three-lens image-shifting mechanism used in commercial image-stabilizing lenses has the potential to satisfy all these requirements. In combination with software designed to track nystagmus, the optical mechanism proved capable of improving visual acuity in 12 normal subjects experiencing simulated two-dimensional nystagmus. Acuity was restored to within an average of 0.12 logMAR (range 0.0-0.22) of the subjects' values without the simulated nystagmus. These results support the feasibility of an assistive device for patients with APN.
OBJECTIVE: To report changes in retinal arterial and venous blood flow pattern in two patients with tumors involving the entire optic nerve. METHODS: Retrospective review of one patient with clinical and neuroimaging characteristics typical of bilateral optic nerve gliomas and one patient with a probable meningioma of the left optic nerve sheath. RESULTS: The optic nerve glioma patient had reduced peak systolic velocity of central retinal arteries bilaterally, while the patient with an optic nerve sheath meningioma had relatively low central retinal artery flow velocity and intermittent blood flow in the central retinal vein on the affected side. CONCLUSIONS: Reduced retinal arterial flow velocities in the setting of optic nerve gliomas may correlate with the presence of optic nerve disease. Phasic blood flow in the central retinal vein with optic nerve sheath meningioma may be the reason that some patients with this tumor develop retinal choroidal venous anastomoses.
The diagnosis of solid benign and malignant tumors presents a unique challenge to all noninvasive imaging modalities. Ultrasound is used in conjunction with mammography to differentiate simple cysts from solid lesions. However, the overlapping appearances of benign and malignant lesions make ultrasound less useful in differentiating solid lesions, resulting in a large number of benign biopsies. Optical tomography using near-infrared diffused light has great potential for imaging functional parameters of 1) tumor hemoglobin concentration, 2) oxygen saturation, and 3) metabolism, as well as other tumor distinguishing characteristics. These parameters can differentiate benign from malignant lesions. However, optical tomography, when used alone, suffers from low spatial resolution and target localization uncertainty due to intensive light scattering. Our aim is to combine diffused light imaging with ultrasound in a novel way for the detection and diagnosis of solid lesions. Initial findings of two early-stage invasive carcinomas, one combined fibroadenoma and fibrocystic change with scattered foci of lobular neoplasia/lobular carcinoma in situ, and 16 benign lesions are reported in this paper. The invasive cancer cases reveal about two-fold greater total hemoglobin concentration (mean 119 micromol) than benign cases (mean 67 micromol), and suggest that the discrimination of benign and malignant breast lesions might be enhanced by this type of achievable optical quantification with ultrasound localization. Furthermore, the small invasive cancers are well localized and have wavelength-dependent appearance in optical absorption maps, whereas the benign lesions appear diffused and relatively wavelength-independent.
BACKGROUND/AIMS: The observation that skin line patterning tends to be disrupted by malignant but not non-malignant skin lesions suggests that this could be used as an aid to lesion differentiation. Since recognised differentiating features can be obtained from the simply-captured white light optical image, the possibility of using such images for skin pattern disruption detection is worth exploring. METHODS: The skin pattern has been extracted from optical images by high-pass filtering and profiles of local line strength variation with the angle estimated using a new consistent high-value profiling technique. The resultant profile images have been analysed using a novel region-based agglomerative clustering technique (mRAC) and also a local variance measurement. A measure based on the relationship between the classification results and an intensity-based segmentation was calculated, and this represented the disruption of the skin line patterning. RESULTS: A set of images containing a variety of histologically confirmed malignant and non-malignant lesions was analysed. The computed textural disruption figure was compared to both the histological diagnosis and to a visual estimate of patterning disruption for each image. It was demonstrated that lesion separation could be achieved by both analysis methods, with a good correlation with visual estimate of disruption and with mRAC providing the best performance. CONCLUSIONS: It was concluded that the acquisition and modelling of skin line patterning from clinical images of skin lesions had been successfully achieved and that the analysis of the resulting data provided an assessment of pattern disruption that is both consistent with visual inspection and effective in presenting information useful for discrimination between melanoma and benign naevi lesion examples.
Optical contrast agents have been widely applied to enhance the sensitivity and specificity of optical imaging with near-infrared (NIR) light. However, because of the overwhelming scattering of light in biological tissues, the spatial resolution of traditional optical imaging degrades drastically as the imaging depth increases. Here, for the first time to our knowledge, we present noninvasive photoacoustic angiography of animal brains in vivo with NIR light and an optical contrast agent. When indocyanine green polyethylene glycol, a novel absorption dye with prolonged clearance, is injected into the circulatory system of a rat, it obviously enhances the absorption contrast between the blood vessels and the background tissues. Because NIR light can penetrate deep into the brain tissues through the skin and skull, we are able to successfully reconstruct the vascular distribution in the rat brain from the photoacoustic signals. On the basis of differential optical absorption with and without contrast enhancement, a photoacoustic angiograph of a rat brain is acquired that matches the anatomical photograph well and exhibits high spatial resolution and a much-reduced background. This new technology demonstrates the potential for dynamic and molecular biomedical imaging.
In vivo imaging of treatment responses at the molecular level could have a significant impact on the speed of drug discovery and ultimately lead to personalized medicine. Strong interest has been shown in developing quantitative fluorescence-based technologies with good molecular specificity and sensitivity for noninvasive 3D imaging through tissues and whole animals. We show herein that tumor response to chemotherapy can be accurately resolved by fluorescence molecular tomography (FMT) with a phosphatidylserine-sensing fluorescent probe based on modified annexins. We observed at least a 10-fold increase of fluorochrome concentration in cyclophosphamide-sensitive tumors and a 7-fold increase of resistant tumors compared with control studies. FMT is an optical imaging technique developed to overcome limitations of commonly used planar illumination methods and demonstrates higher quantification accuracy validated by histology. It is further shown that a 3-fold variation in background absorption heterogeneity may yield 100% errors in planar imaging but only 20% error in FMT, thus confirming tomographic imaging as a preferred tool for quantitative investigations of fluorescent probes in tissues. Tomographic approaches are found essential for small-animal optical imaging and are potentially well suited for clinical drug development and monitoring.
We present a spatially selective irradiation method for laser treatment of biological surfaces. The purpose is to irradiate only the pathological targets and to preserve the healthy surrounding parts. We are interested here in the optical arrangement of the device and in the evaluation of its physical limitations before it is used for medical purposes. The method uses real-time image analysis applied to the video imagery of the surface to be treated. The result of the image analysis generates the control signal to a spatial light modulator, allowing us to project an adequate repartition of laser irradiance on the surface to be treated. Experimental results demonstrate a spatial selectivity of approximately 10 microm for a 6.7 mm x 5 mm field. The optical system has no moving parts (unlike galvanometric scanners) and is able to irradiate multiple targets simultaneously with different doses.
PURPOSE: To determine if different expression levels of tumor cathepsin-B activity in well differentiated and undifferentiated breast cancers could be revealed in vivo with optical imaging. MATERIALS AND METHODS: A well differentiated human breast cancer (BT20, n = 8) and a highly invasive metastatic human breast cancer (DU4475, n = 8) were implanted orthotopically in athymic nude mice. Tumor-bearing animals were examined in vivo with near-infrared fluorescence (NIRF) imaging 24 hours after intravenous injection of an enzyme-sensing imaging probe. Immunohistochemistry, Western blotting (on cells and whole tumor samples), and correlative fluorescence microscopy were performed. RESULTS: Both types of breast cancers activated the NIRF probe so that tumors became readily detectable. However, in tumors of equal size, there was a 1.5-fold higher fluorescence signal in the highly invasive breast cancer (861 arbitrary units +/- 88) compared with the well differentiated lesion (566 arbitrary units +/- 36, P <.01). Western blotting confirmed a higher cathepsin-B protein content in the highly invasive breast cancer (DU4475) of about 1.4-fold (whole tumor samples) to 1.7-fold (cells). Immunohistochemistry and fluorescence microscopy findings confirmed the imaging findings. CONCLUSION: Cathepsin-B enzyme activity can be determined in vivo with NIRF optical imaging, while differences in tumoral expression may correlate with tumor aggressiveness.