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PubMed2004

Extraction of optical scattering parameters and attenuation compensation in optical coherence tomography images of multilayered tissue structures.

A recently developed analytical optical coherence tomography (OCT) model [Thrane et al., J. Opt. Soc. Am. A 17, 484 (2000)] allows the extraction of optical scattering parameters from OCT images, thereby permitting attenuation compensation in those images. By expanding this theoretical model, we have developed a new method for extracting optical scattering parameters from multilayered tissue structures in vivo. To verify this, we used a Monte Carlo (MC) OCT model as a numerical phantom to simulate the OCT signal for heterogeneous multilayered tissue. Excellent agreement between the extracted values of the optical scattering properties of the different layers and the corresponding input reference values of the MC simulation was obtained, which demonstrates the feasibility of the method for in vivo applications. This is to our knowledge the first time such verification has been obtained, and the results hold promise for expanding the functional imaging capabilities of OCT.

Algorithms↗
PubMed2003

Multiphoton endoscopy.

Despite widespread use of multiphoton fluorescence microscopy, development of endoscopes for nonlinear optical imaging has been stymied by the degradation of ultrashort excitation pulses that occurs within optical fiber as a result of the combined effects of group-velocity dispersion and self-phase modulation. We introduce microendoscopes (350-1000 microm in diameter) based on gradient-index microlenses that effectively eliminate self-phase modulation within the endoscope. Laser-scanning multiphoton fluorescence endoscopy exhibits micrometer-scale resolution. We used multiphoton endoscopes to image fluorescently labeled neurons and dendrites.

Animals↗
PubMed2002

Characteristics of topographical heterogeneity of articular cartilage over the joint surface of a humeral head.

OBJECTIVE: To investigate the topographical variations among the morphological and physical properties of articular cartilage over a canine humeral head. DESIGN: Nine side-by-side specimens from a canine humeral head were used in the combined polarized light microscopy (PLM) and microscopic magnetic resonance imaging (muMRI). Sixteen images were acquired from each specimen in muMRI. Subsequently, ten to fourteen histological slices were prepared at the location of the muMRI slice from each specimen. Four optical images were acquired from each histological slice. Using these images, the images of the T(2) relaxation, the optical retardation, and the collagen-fiber orientation were constructed in two dimensions. RESULTS: Along the medial/lateral direction of the humeral head, we have observed a number of topographical variations. These include the total thickness of the cartilage tissue, the thickness of the individual histological zones, the depth of the minimum optical retardation in the tissue, and the depth of the maximum T(2) relaxation in the tissue. We found that the depth of the minimum retardation from PLM occurs at the geometric middle point of the transitional zone in histology, and that the depth of the maximum T(2) relaxation from muMRI is closely correlated with the minimum retardation depth in PLM. In addition, although the thickness of the total tissue increases approximately by the same amount on both the lateral and medial sides of the joint, a slight asymmetry exists between the measurements from the medial and lateral halves of the humerus. CONCLUSIONS: Cartilage from different sites over a single joint could have different properties, possibly the consequence of the variation in load-bearing status. Because of these topographical variations, one must keep in mind that a different sampling site within a relatively small area of the same joint may significantly influence the results of the study.

Animals↗
PubMed2005

Deconvolution methods for mitigation of transverse blurring in optical coherence tomography.

Imaging resolution in optical coherence tomography (OCT) is a key determinant for acquiring clinically useful optical biopsies of tissues. In contrast to light or confocal microscopy, the axial and transverse resolutions in OCT are independent and each can be analyzed individually. A method for mitigating transverse blurring and the apparent loss of transverse resolution in OCT by means of Gaussian beam deconvolution is presented. Such a method provides better representation of a specimen by using known physical parameters of a lens. To implement this method, deconvolution algorithms based on a focal-dependent kernel are investigated. First, the direct inverse problem is investigated using two types of regularization, truncated singular value decomposition, and Tikhonov. Second, an iterative expectation maximization algorithm, the Richardson-Lucy algorithm, with a beam-width-dependent iteration scheme is developed. A dynamically iterative Richardson-Lucy algorithm can reduce transverse blurring by providing an improvement in the transverse point-spread-function for sparse scattering samples in regions up to two times larger than the confocal region of the lens. These deblurring improvements inside and outside of the confocal region, which are validated experimentally, are possible without introducing new optical imaging hardware or acquiring multiple images of the same specimen. Implementation of this method in sparse scattering specimens, such as engineered tissues, has the potential to improve cellular detection and categorization.

Algorithms↗
PubMed1991

Leber's hereditary optic neuropathy and complex I deficiency in muscle.

We investigated a family with Leber's hereditary optic neuropathy in which affected individuals were homoplasmic for the point mutation of the NADH-dehydrogenase 4 gene of mitochondrial DNA, described by Wallace and colleagues in 1988. The proband had bilateral optic atrophy, tremor, dystonia, and sharply defined lesions in the putamen on magnetic resonance images. Optic atrophy was found in another 3 of 13 investigated relatives on the maternal side. Additional neurological signs were found but only in patients with optic neuropathy. The morphological appearance and the respiratory chain function of muscle tissue were investigated in the proband, his mother, and 3 siblings. Polarographic measurements revealed complex I deficiency in the 5 investigated subjects. Morphological changes of mitochondria were found in 4 of these subjects. There was no decrease in complex I activity measured as NADH ferricyanide reductase or rotenone-sensitive NADH cytochrome c reductase activities. In other cases with complex I deficiency, good agreement between polarographic and spectrophotometric measurements was found. This study showed that there is decreased activity of complex I of the respiratory chain in muscle and that cerebral striatal lesions occur in Leber's hereditary optic neuropathy with the NADH-dehydrogenase 4 gene point mutation.

Adolescent↗
PubMed2003

Real time non-invasive imaging of receptor-ligand interactions in vivo.

Non-invasive longitudinal detection and evaluation of gene expression in living animals can provide investigators with an understanding of the ontogeny of a gene's biological function(s). Currently, mouse model systems are used to optimize magnetic resonance imaging (MRI), positron emission tomography (PET), single-photon emission computed tomography (SPECT), and optical imaging modalities to detect gene expression and protein function. These molecular imaging strategies are being developed to assess tumor growth and the tumor microenvironment. In addition, pre-labeling of progenitor cells can provide invaluable information about the developmental lineage of stem cells both in organogenesis and tumorigenesis. The feasibility of this approach has been extensively tested by targeting of endogenous tumor cell receptors with labeled ligand (or ligand analog) reporters and targeting enzymes with labeled substrate (or substrate analog). We will primarily discuss MRI, PET, and SPECT imaging of cell surface receptors and the feasibility of non-invasive imaging of gene expression using the tumor microenvironment (e.g., hypoxia) as a conditional regulator of gene expression.

Animals↗
PubMed2006

Schlieren visualization of ultrasonic wave fields with high spatial resolution.

The visualization of ultrasonic wave fields in optically transparent liquids using the acousto-optic interaction is a well proven tool for the experimental investigation of wave propagation including wave field interaction effects with certain discontinuities and obstacles like reflection, refraction, and diffraction effects as well as for transducer testing and design. For high resolution visualization of wave fields including pulsed waveforms, pulsed light sources and sensitive optical imaging sensors with certain specifications are needed. In this paper the technical requirements of optical and electronic components for high resolution visualization of ultrasound wave fields will be presented. Also, specifications and operation results of a new designed, inexpensive Schlieren optical system will be presented, which is capable of pulsed wave field visualization in the MHz frequency range. The spatial resolution is high enough, not only for accurate beam shape and wave pattern visualization, but also for a gray-scaled display of wave amplitudes including amplitude zero crossings in ultrasound pulses. Consequently, ultrasonic wavelengths can be visualized quantitatively as well as wavelength changes of the ultrasound pulses while traveling through transparent media with different sound velocities. Results to be presented will include 2 MHz and 10 MHz experiments using single transducers as well as linear arrays of commercial medical scanners during their standard operation showing the system beamforming characteristics.

Journal Article↗
PubMed2001

Light rays and imaging in wave optics.

An interpretation of focusing and image formation on scattering of electromagnetic waves by a dielectric cylinder (a cylindrical lens) is proposed on the basis of the full Maxwell theory. It is centered on analysis of the behavior of integral curves of the Poynting vector here called wave rays. These wave rays cannot intersect so that the focusing and imaging spots are identified with regions of high flow concentration. Two-dimensional examples of wave rays and wave fronts in the scattering of plane and cylindrical electromagnetic waves as well as of Gaussian beams by a dielectric cylinder derived from rigorous solution of the Maxwell equations for incident waves perpendicular to and uniform along the scatterer are given. Their qualitative comparison with geometrical and diffraction approximations are provided. Fixed points and vortex structure of the Poynting flow are investigated. An example of (Gaussian-beam) scattering with transparent multiple internal reflections and multiple wave splitting is given.

Journal Article↗
PubMedDate not supplied

Targeted molecular imaging.

Molecular imaging aims to visualize the cellular and molecular processes occurring in living tissues, and for the imaging of specific molecules in vivo, the development of reporter probes and dedicated imaging equipment is most important. Reporter genes can be used to monitor the delivery and magnitude of therapeutic gene transfer, and the time variation involved. Imaging technologies such as micro-PET, SPECT, MRI and CT, as well as optical imaging systems, are able to non-invasively detect, measure, and report the simultaneous expression of multiple meaningful genes. It is believed that recent advances in reporter probes, imaging technologies and gene transfer strategies will enhance the effectiveness of gene therapy trials.

Animals↗
PubMedDate not supplied

[Improvement of the contrast of electron microscope images by the optical shadow method].

Amplitude contrast of images of weak phase and amplitude objects can be strengthened almost twice as compared to standard light-pole contrast by means of shadow method of image formation without special contrasting of the objects. In spite of the contour effects (electron-microscopic attenuation) appearing at image formation by asymmetrical shadow method, the quantitative interpretation of such image is quite possible. The symmetrical shadow image (image with cone illumination) is more complexly realized than the asymmetric one, but it has some additional advantages. Particularly efficient suppression of background noises is there possible. Several symmetrical shadow images can be synthetized by the differential method with or without colour coding into the final image with the signal/noise ratio increased by an order.

Animals↗
PubMed2003

Image reconstruction of optical attenuation coefficient variation in biological tissues.

A procedure for non-invasive imaging of the optical attenuation coefficient variation of in vivo thick organs/tissues is developed. The laser back-scattered surface profiles at various locations of human forearm, by multi-probe reflectometer, are measured. These profiles are matched by iterative procedure, with that as obtained by Monte Carlo simulation and the corresponding values of attenuation coefficient (equal to the sum of absorption and reduced scattering coefficients) are determined. By interpolation of this data a 100 x 100 grid is constructed and after median filtering of this data a color-coded image of the variability of the optical attenuation coefficient of the forearm is obtained. These images in different subjects show variation due to change in overall tissue composition and blood pooling. This non-invasive imaging procedure may help in identifying the diseased affected regions in healthy tissues and in application of photodynamic therapy.

Diagnostic Imaging↗
PubMed2002

MR imaging of the globe and optic nerve.

MR imaging is an important tool for the evaluation of the optic nerve and globe. A working knowledge of the clinical questions an ophthalmologist faces helps radiologists optimize MR imaging protocols and focus their attention on the important clinical issues.

Eye↗
PubMed2006

Partial coherence and other optical delicacies of lepidopteran superposition eyes.

Superposition eyes are generally thought to function ideally when the eye is spherical and with rhabdom tips in the focal plane of the imaging optics of facet lenses and crystalline cones. Anatomical data as well as direct optical measurements demonstrate that the superposition eyes of moths and skippers often deviate severely from the expected ideal case. Part of the deviation has been attributed to diffraction at the single facet lens, which was taken to be an essential limit to spatial resolution, because light traveling through different facet lenses was assumed to be incoherent. By considering the two-dimensional facet lens lattice, it is here demonstrated that many facets within a superposition aperture transmit coherent light, allowing a much sharper image than possible with single facet lens diffraction. Partial coherence therefore is an important aspect of superposition imaging. It is argued that broadening of the photoreceptor acceptance angles occurs because of optical errors in the facet lens-crystalline cone system other than diffraction. The transmittance of the superposition aperture of moths and skippers is improved by the corneal nipple arrays of the facet lenses, but quantitative assessment shows that the effect is minor.

Animals↗
PubMedDate not supplied

Preparation of a cathepsin D sensitive near-infrared fluorescence probe for imaging.

A variety of proteases are overexpressed or activated during pathogenesis and represent important targets for therapeutic drugs. We have previously shown that optical imaging probes sensitive in the near-infrared fluorescence (NIRF) spectrum can be used for in vivo imaging of enzyme activity. In the current study, we show that these probes can be designed with specificity for specific enzymes, for example, cathepsin D which is known to be overexpressed in many tumors. A NIR cyanine fluorochrome served as the optical reporter and was attached to the amino terminal of an 11 amino acid peptide sequence with specificity for cathepsin D. The peptides were subsequently attached to a synthetic graft copolymer for efficient tumoral delivery. The close spatial proximity of the multiple fluorochromes resulted in quenching of fluorescence in the bound state. A 350-fold signal amplification was observed post cleavage during in vitro testing. Cell culture experiments using a rodent tumor cell line stably transfected with human cathepsin D confirmed enzyme specific activation within cells. This sequence but not a scrambled control sequence showed enzyme specificity in vitro. We conclude that activatable NIRF optical probes can be synthesized to potentially probe for specific enzymes in living organisms.

Animals↗
PubMed1981

Computer-enhanced video microscopy: digitally processed microscope images can be produced in real time.

Digital processing techniques can be used to greatly enhance the available information in an optical image. Although this technology has been routinely used in many fields for a number of years, little application of digital image-processing techniques have been made toward analysis and enhancement of the types of images seen most often by the research biologist. We describe here a computer-based video microscope system that is capable of performing extensive manipulation and enhancement of microscope images in real time. The types of manipulations possible with these techniques greatly surpass the enhancement capabilities of photographic or video techniques alone. The speed and flexibility of this system enables experimental manipulation of the microscopic specimen based on its live processed image. These features greatly extend the power and versatility of the light microscope.

Animals↗
PubMed1999

Convergent, self-encoded bead sensor arrays in the design of an artificial nose.

We report a new approach to designing an artificial nose based on high-density optical arrays that directly incorporate a number of structural and operational features of the olfactory system. The arrays are comprised of thousands of microsphere (bead) sensors, each belonging to a discrete class, randomly dispersed across the face of an etched optical imaging fiber. Beads are recognized and classified after array assembly by their unique, "self-encoded" response pattern to a selected vapor pulse. The high degree of redundancy built into the array parallels that found in nature and affords new opportunities for chemical-sensor signal amplification. Since each bead is independently addressable through its own light channel, it is possible to combine responses from same-type beads randomly distributed throughout the array in a manner reminiscent of the sensory-neuron convergence observed in the mammalian olfactory system. Signal-to-noise improvements of approximately n1/2 have been achieved using this method.

Artificial Organs↗
PubMed2004

Optic nerve size evaluated by magnetic resonance imaging in children with optic nerve hypoplasia, multiple pituitary hormone deficiency, isolated growth hormone deficiency, and idiopathic short stature.

OBJECTIVE: To objectively define criteria for intracranial optic nerve (ON) size in ON hypoplasia (ONH) on magnetic resonance imaging (MRI) scans. STUDY DESIGN: Intracranial ON sizes from MRI were compared between 46 children with ONH diagnosed by ophthalmoscopy (group 1, isolated ONH, 8 children; and group 2, ONH associated with abnormalities of the hypothalamic-pituitary axis and septum pellucidum, 38 children) and children with multiple pituitary hormone deficiency (group 3, multiple pituitary hormone deficiency, 14 children), isolated growth hormone deficiency (group 4, isolated growth hormone deficiency, 15 children), and idiopathic short stature (group 5, idiopathic short stature, 10 children). Intracranial ON size was determined by the cross-sectional area, calculated as [pi x (1/2) height x (1/2) width]. RESULTS: Groups 1 and 2 had lower intracranial ON size than did groups 3, 4, and 5 (P < .001). No patients in groups 3 through 5 who had MRI after 12 months of age (when 95% adult size of ONs is attained) had ONs <2.9 mm 2 . Visual acuity correlated significantly with ON size (P < .01). CONCLUSIONS: Magnetic resonance imaging of the ONs with cross-sectional area <2.9 mm 2 in a short child more than 12 months of age, with or without hypothalamic-pituitary axis abnormalities, confirms the clinical diagnosis of ONH.

Adolescent↗