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PubMed2004

Optical imaging of the heart.

Optical techniques have revolutionized the investigation of cardiac cellular physiology and advanced our understanding of basic mechanisms of electrical activity, calcium homeostasis, and metabolism. Although optical methods are widely accepted and have been at the forefront of scientific discoveries, they have been primarily applied at cellular and subcellular levels and considerably less to whole heart organ physiology. Numerous technical difficulties had to be overcome to dynamically map physiological processes in intact hearts by optical methods. Problems of contraction artifacts, cellular heterogeneities, spatial and temporal resolution, limitations of surface images, depth-of-field, and need for large fields of view (ranging from 2x2 mm2 to 3x3 cm2) have all led to the development of new devices and optical probes to monitor physiological parameters in intact hearts. This review aims to provide a critical overview of current approaches, their contributions to the field of cardiac electrophysiology, and future directions of various optical imaging modalities as applied to cardiac physiology at organ and tissue levels.

Action Potentials↗
PubMed1996

Laser-scanned agarose gel sections for radiation field mapping.

PURPOSE: A dedicated laser scanning system has been applied to the measurement of optical density change in ferrous sulphate-xylenol orange dye-agarose gel mixtures for applications in radiation field mapping. METHODS AND MATERIALS: The gels were poured, irradiated, and optically scanned in Lucite casts providing a uniform gel section thickness of 1 cm. The gel sections were optically scanned both pre- and postirradiation to determine the optical density change in the gel and consequently the absorbed dose. Irradiations of the gel sections were performed with an orthovoltage unit and a linear accelerator. RESULTS: For linear dose response up to 10 Gy, an appropriate gel mixture was found to be 0.4 mM Fe2+, 0.2mM xylenol orange dye, 25 mM sulphuric acid, and 1% by weight agarose gel with a mixing temperature of 60 degrees C. In dosimetry of a 20 x 10 cm 6 MV wedged x-ray field, good agreement in terms of relative dose was found between the gel values and ionization chamber readings. However, in repeated experiments with the gel dosimeter involving calculation of absolute dose at various points in the wedged field, variations in absolute dose measurements of up to +/- 5% were observed. CONCLUSION: The dosimetry technique involving laser scanning of agarose gel sections has potential for further applications in radiotherapy dosimetry.

Dose-Response Relationship, Radiation↗
PubMed1991

Identification of Ser-55 as a major protein kinase A phosphorylation site on the 70-kDa subunit of neurofilaments. Early turnover during axonal transport.

The 70-kDa neurofilament protein subunit (NF-L) is phosphorylated in vivo on at least three sites (L1 to L3) (Sihag, R. K. and Nixon, R. A. (1989) J. Biol. Chem. 264, 457-464). The turnover of phosphate groups on NF-L during axonal transport was determined after the neurofilaments in retinal ganglion cells were phosphorylated in vivo by injecting mice intravitreally with [32P]orthophosphate. Two-dimensional phosphopeptide maps of NF-L from optic axons of mice 10 to 90 h after injection showed that radiolabel decreased faster from peptides L2 and L3 than from L1 as neurofilaments were transported. To identify phosphorylation sites on peptide L2, axonal cytoskeletons were phosphorylated by protein kinase A in the presence of heparin. After the isolated NF-L subunits were digested with alpha-chymotrypsin, 32P-peptides were separated by high performance liquid chromatography on a reverse-phase C8 column. Two-dimensional peptide mapping showed that the alpha-chymotrypsin 32P-peptide accepting most of the phosphates from protein kinase A migrated identically with the in vivo-labeled phosphopeptide L2. The sequence of this peptide (S-V-R-R-S-Y) analyzed by automated Edman degradation corresponded to amino acid residues 51-56 of the NF-L sequence. A synthetic 13-mer (S-L-S-V-R-R-S-Y-S-S-S-S-G) corresponding to amino acid residues 49-61 of NF-L was also phosphorylated by protein kinase A. alpha-Chymotryptic digestion of the 13-mer generated a peptide which contained most of the phosphates and co-migrated with the phosphopeptide L2 on two-dimensional phosphopeptide maps. Edman degradation of the phosphorylated 13-mer identified serine residue 55 which is located within a consensus phosphorylation sequence for protein kinase A as the major site of phosphorylation. Since protein kinase A-mediated phosphorylation influences intermediate filament assembly/disassembly in vitro, we propose that the phosphopeptide L2 region is a neurofilament-assembly domain and that the cycle of phosphorylation and dephosphorylation of Ser-55 on NF-L, which occurs relatively early after subunit synthesis in vivo, regulaaes a step in neurofilament assembly or initial interactions during axonal transport.

Amino Acid Sequence↗
PubMed2003

Intensity and phase mapping of guided light in LiNbO3 waveguides with an interferometric near-field scanning optical microscope.

The design and implementation of a phase-sensitive near-field scanning optical microscope incorporating both heterodyne interferometric detection and a phase feedback mechanism are described. Using this microscope we demonstrate a new method for measuring the effective index of the guided mode of a waveguide from the phase images. Two types of LiNbO3 waveguide, defined by titanium diffusion or annealed proton exchange, were studied. Both the profile and the effective index of the mode were measured experimentally. For titanium-diffused waveguides, both agree well with values determined from numerical simulation. In annealed proton-exchanged waveguides that contain periodically poled domains, we find evidence for backreflection from the boundaries between neighboring regions of opposite pole directions, which could result in transmission loss in this type of waveguide.

Journal Article↗
PubMed1975

[Retinal projections in Caiman sclerops. Autoradiographic study].

The retinal projections in Caiman sclerops have been re-examined using the radio-autographic method for tracing axonal pathways. Following the intraocular injection of a mixture of [3H]proline and [3H]fucose, label transported in the rapid phase of axonal flow was found only on controlateral side to the injection in the nuceli, supra-opticus, ovalis, geniculatus lateralis pars dorsalis and pars ventralis, geniculatus pretectalis, lentiformis mesencephali, griseus tectalis, opticus tegmenti and in the superficial layers of the tectum opticum (layers 1 to 6). Label transported in the slow phase confirms the absence of an ipsilateral optic component. No controlateral retinofugal fasicule crossing over in the posterior commissure as previously described in this species using the Nauta method (3) was observed. In comparison with degeneration study in Caiman sclerops (3) the radio-autographic results demonstrated more accurately the sites of termination of retinofugal fibers.

Alligators and Crocodiles↗
PubMed1998

Visual hemifield mapping using transcranial magnetic stimulation coregistered with cortical surfaces derived from magnetic resonance images.

The perception of a visual stimulus can be inhibited by occipital transcranial magnetic stimulation. This visual suppression effect has been attributed to disruption in the cortical gray matter of primary visual cortex or in the fiber tracts leading to V1 from the thalamus. However, others have suggested that the visual suppression effect is caused by disruption in secondary visual cortex. Here the authors used a figure-eight coil, which produces a focal magnetic field, and a Quadropulse stimulator to produce visual suppression contralateral to the stimulated hemisphere in five normal volunteer subjects. The authors coregistered the stimulation sites with magnetic resonance images in these same subjects using optical digitization. The stimulation sites were mapped onto the surface of the occipital lobes in three-dimensional reconstructions of the cortical surface to show the distribution of the visual suppression effect. The results were consistent with disruption of secondary visual cortical areas.

Adult↗
PubMed2003

Diffusion tractography based group mapping of major white-matter pathways in the human brain.

Tractography uses diffusion tensor imaging data to trace white matter pathways in vivo within the brain. We have constructed group maps that represent three major white matter tracts-the anterior callosal fibers, optic radiations, and pyramidal tracts-in a group of 21 volunteers. For each individual tract the fast marching tractography (FMT) algorithm was used to generate a VSC (voxel scale connectivity) map in native space. Using SPM99 these maps were transformed into a standard reference frame and three group mapping techniques were investigated: the first averaged the individual VSC maps, the second produced maps that demonstrate intersubject tract variability and degree of overlap, and the third used an SPM analysis to construct a statistical image that represents the group effect. The group maps reconstructed for each tract under investigation conform well to known anatomy and are consistent with data derived from postmortem human brains. Greater intersubject variability is found around the terminal projections of the tracts adjacent to cerebral cortex, whereas the "core" of each tract is characterized by lower variability. No significant differences were found between the left and right side of the pyramidal tracts and optic radiations. The group mapping techniques utilize the VSC maps in different but complementary ways. In the future, group mapping could investigate in vivo white matter differences between normal subjects and patients affected by neurological and psychiatric diseases.

Adult↗
PubMedDate not supplied

The apparent diffusion coefficient of water in gray and white matter of the infant brain.

PURPOSE: The purpose was to obtain normal values of the apparent diffusion coefficient (ADC) in the infant brain and to compare ADC maps with T1- and T2-weighted images. METHOD: Diffusion was measured in nine infants with an ECG-gated SE sequence compensated for first-order motion. One axial slice at the basal ganglia level was investigated with the diffusion-encoding gradients in the slice-selection direction. RESULTS: On ADC maps, the corpus callosum and the optic radiations appeared dark before the onset of myelination, and the crus posterior of the internal capsule could be visualized before it appeared on T1- or T2-weighted images. In gray and white matter, the mean ADC ranged from 0.95 x 10(-9) to 1.76 x 10(-9) m2/s. In the frontal and occipital white matter, in the genu corporis callosi, and in the lentiform nucleus, the ADC decreased with increasing age. The cortex/white matter ratio of the ADC increased with age and approached 1 at the age of 30 weeks. CONCLUSION: ADC maps add information to the T1 and T2 images about the size and course of unmyelinated as well as myelinated tracts in the immature brain.

Brain↗
PubMed1980

Constant occurrence of an ipsilateral retino-tectal projection in the axolotl (Ambystoma mexicanum) revealed by horseradish peroxidase tracing.

Cut optic nerves were filled orthogradely with horseradish peroxidase (HRP) to reveal contralateral and ipsilateral retino-tectal projections in the axolotl, by light and electron microscopy. The ipsilateral projection is small but consistently present. It is concentrated in the rostral tectum particularly the dorsomedial and ventrolateral areas. The results are compared with findings in other vertebrates.

Ambystoma mexicanum↗
PubMedDate not supplied

Optical release of caged glutamate for stimulation of neurons in the in vitro slice preparation.

Optical stimulation techniques prove useful to map functional inputs in the in vitro brain slice preparation: Glutamate released by a focused beam of UV light induces action potentials, which can be detected in postsynaptic neurons. The direct activation effect is influenced by factors such as compound concentration, focus depth, light absorption in the tissue, and sensitivity of different neuronal domains. We analyze information derived from direct stimulation experiments in slices from rat barrel cortex and construct a computational model of a layer V pyramidal neuron that reproduces the experimental findings. The model predictions concerning the influence of focus depth on input maps and action potential generation are investigated further in subsequent experiments where the focus depth of a high-numerical-aperture lens is systematically varied. With our setup flashes from a xenon light source can activate neuronal compartments to a depth of 200 mum below the surface of the slice. The response amplitude is influenced both by tissue depth and focus plane. Specific somatodendritic structures can be targeted as the probability of action potential induction falls off exponentially with distance. Somata and primary apical dendrites are most sensitive to uncaged glutamate with locally increased sensitivity on proximal apical dendrites. We conclude that optical stimulation can be targeted with high precision.

Action Potentials↗
PubMed1997

Representation of sound localization cues in the auditory thalamus of the barn owl.

Barn owls can localize a sound source using either the map of auditory space contained in the optic tectum or the auditory forebrain. The auditory thalamus, nucleus ovoidalis (N.Ov), is situated between these two auditory areas, and its inactivation precludes the use of the auditory forebrain for sound localization. We examined the sources of inputs to the N.Ov as well as their patterns of termination within the nucleus. We also examined the response of single neurons within the N.Ov to tonal stimuli and sound localization cues. Afferents to the N.Ov originated with a diffuse population of neurons located bilaterally within the lateral shell, core, and medial shell subdivisions of the central nucleus of the inferior colliculus. Additional afferent input originated from the ipsilateral ventral nucleus of the lateral lemniscus. No afferent input was provided to the N.Ov from the external nucleus of the inferior colliculus or the optic tectum. The N.Ov was tonotopically organized with high frequencies represented dorsally and low frequencies ventrally. Although neurons in the N.Ov responded to localization cues, there was no apparent topographic mapping of these cues within the nucleus, in contrast to the tectal pathway. However, nearly all possible types of binaural response to sound localization cues were represented. These findings suggest that in the thalamo-telencephalic auditory pathway, sound localization is subserved by a nontopographic representation of auditory space.

Acoustic Stimulation↗
PubMed1996

Disproportionate distribution of field potentials across the toad's tectal visual map in response to diffuse light ON and OFF stimulations.

In toads Bufo marinus and Bufo bufo spinosus, field potentials (FPs) were recorded from the surface of the optic tectum at different sites of the visual map in response to a sudden diffuse darkening (OFF) and lightening (ON) of the visual field of the contralateral eye. The OFF and ON responses were differently pronounced or even failed to occur. The latency of the former was significantly less than the one of the latter. FP amplitudes of the OFF and ON responses were strongest in the representation of a horizonto-superior anterio-lateral portion of the visual field and weakest toward the posterior field of vision. This phenomenon suggests various interpretations for subsequent experiments.

Animals↗
PubMed1988

Control of tectal cell number during larval development in Rana pipiens.

Cell production and cell deaths were determined in larval Rana pipiens both in control tecta and in tecta following unilateral eyeball removal in embryos and larvae. Such enucleations produce significantly reduced rates of cell division in the contralateral tecta for virtually the entire larval period (confirming studies with enucleation almost exclusively performed in embryos--Kollros: J. Exp. Zool. 123:153-187, '53, and J. Comp. Neurol. 205:171-178, '82). Significant numbers of cell deaths in all nonependymal tectal cell layers were also observed. Control cell division rates peak at stage X, while cell death peaks are reached in stages XIII-XX. Overall, about 10(6) nonependymal cells are produced in control tecta, and about 350,000 of them die by the end of metamorphosis. Control of cell numbers following enucleation is shown to depend mainly on reductions in cell division rates when the operation occurs early in development and mainly on increases in cell death rates when the operation occurs late in larval life. Such increases in death rates are invariably present within 1 day of the operation whereas the reduced division rates ordinarily require several more days to be seen. The modified rates, both of cell divisions and cell death, are limited to tectal areas to which optic nerve fibers have already extended. Maps of the positions of tectal cell divisions in many larval stages provide the basis for modifying the current dogma that tectal formation occurs as a series of newly formed mediocaudal wedges pushing previously produced wedges rostrolaterad. All such "old" wedges receive substantial cell additions for many stages, with the rate of addition decreasing rostrad earlier than caudad.

Animals↗
PubMed2002

Plasticity of orientation preference maps in the visual cortex of adult cats.

In contrast to the high degree of experience-dependent plasticity usually exhibited by cortical representational maps, a number of experiments performed in visual cortex suggest that the basic layout of orientation preference maps is only barely susceptible to activity-dependent modifications. In fact, most of what we know about activity-dependent plasticity in adults comes from experiments in somatosensory, auditory, or motor cortex. Applying a stimulation protocol that has been proven highly effective in other cortical areas, we demonstrate here that enforced synchronous cortical activity induces major changes of orientation preference maps (OPMs) in adult cats. Combining optical imaging of intrinsic signals and electrophysiological single-cell recordings, we show that a few hours of intracortical microstimulation (ICMS) lead to an enlargement of the cortical representational zone at the ICMS site and an extensive restructuring of the entire OPM layout up to several millimeters away, paralleled by dramatic changes of pinwheel numbers and locations. At the single-cell level, we found that the preferred orientation was shifted toward the orientation of the ICMS site over a region of up to 4 mm. Our results show that manipulating the synchronicity of cortical activity locally without invoking training, attention, or reinforcement, OPMs undergo large-scale reorganization reminiscent of plastic changes observed for nonvisual cortical maps. However, changes were much more widespread and enduring. Such large-scale restructuring of the visual cortical networks indicates a substantial capability for activity-dependent plasticity of adult visual cortex and may provide the basis for cognitive learning processes.

Age Factors↗
PubMed1990

Restoration of the plasticity of binocular maps by NMDA after the critical period in Xenopus.

Visual input during a critical period of development plays a major role in the establishment of orderly connections in the developing visual system. In Xenopus laevis, the matching of visual maps from the two eyes to the optic tectum depends on binocular visual input during the critical period, which extends from late tadpole to early juvenile stages. Alterations in eye position, which produce a mismatch of the tectal maps, normally evoke a compensatory adjustment in the map of the ipsilateral eye only during the critical period. However, continuous application of the glutamate receptor agonist N-methyl-D-aspartate (NMDA) after the normal end of the critical period restores this ability to realign the visual map.

Animals↗
PubMed2006

Theoretical near-field optical properties of branched plasmonic nanoparticle networks.

Extended branched networks of single-nanoparticle chains have recently been self-assembled from colloidal suspensions. In particular, gold nanoparticle linear chains and complex chain networks have revealed unique signatures of plasmon modes in their extinction spectra. In this Letter, we investigate theoretically their near-field optical properties and show that a real space mapping of these modes can be achieved with a photon scanning tunneling microscope setup. A distinct subwavelength patterning of the optical near-field gives rise to well-resolved photon scanning tunneling microscope images that can be used to identify the network segments able to efficiently carry optical energy.

Journal Article↗
PubMedDate not supplied

Combined diffuse optical spectroscopy and contrast-enhanced magnetic resonance imaging for monitoring breast cancer neoadjuvant chemotherapy: a case study.

Monitoring tumor response to therapy can enable assessment of treatment efficacy, maximizing patient outcome and survival. We employ a noninvasive, handheld laser breast scanner (LBS) based on broadband diffuse optical spectroscopy (DOS) in conjunction with contrast-enhanced magnetic resonance imaging (cMRI) to assess tumor response to presurgical neoadjuvant chemotherapy. DOS and cMRI scans are performed after the first and fourth cycles of a doxorubicin/cyclophosphamide regimen in a patient with invasive ductal carcinoma. DOS measurements are used to quantify bulk tissue optical and physiological parameters, which are mapped to T2- and T1-weighted cMRI images. Initial DOS measurements show high tumor/normal contrast in total hemoglobin concentration (THC, 56+/-7 versus 27+/-4 microM) and water fraction (81.4+/-1% versus 24+/-3%) colocalized with regions of strongly enhancing T2-weighted and cMRI signals. After the fourth cycle of chemotherapy, we observe decreases in peak MRI contrast-enhancement values (37.6%) and apparent lesion volume (21.9 versus 13.7 cm3), which corresponds to physiological changes measured by DOS, including a 20 to 25% reduction in the spatial extent of the tumor and a 38.7% drop in mean total hemoglobin content (THC, 41.6 versus 23.4 microM). These data provide in vivo validation of the accuracy of broadband DOS and the sensitivity of optical methods to changes in tumor physiology.

Antineoplastic Combined Chemotherapy Protocols↗