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At least 883 records · Page 49Linked to original sources
PubMed2005

Spatiotemporal activity patterns during respiratory rhythmogenesis in the rat ventrolateral medulla.

One of the most important brain rhythms is that which generates involuntary breathing movements. The lower brain stem contains neural circuitry for respiratory rhythm generation in mammals. To date, microsectioning and selective lesioning studies have revealed anatomical regions necessary for respiratory rhythmogenesis. Although respiratory neurons distributed within these regions can be identified by their firing patterns in different phases of the respiratory cycle, conventional electrophysiology techniques have limited the study of spatial organization within this network. Optical imaging techniques offer the potential for monitoring the spatiotemporal activity of large groups of neurons simultaneously. Using high-speed voltage-sensitive dye imaging and spatial correlation analysis in an arterially perfused in situ preparation of the juvenile rat, we determined the spatial distribution of respiratory neuronal activity in a region of the ventrolateral respiratory group containing the pre-Bötzinger complex (pBC) during spontaneous eupneic breathing. While distinctly pre- and postinspiratory-related responses were spatially localizable on length scales less than 100 microm, we found the studied area on whole exhibited a spatial mixture of phase-spanning and postinspiratory-related activity. Additionally, optical recordings revealed significant widespread hyperpolarization, suggesting inhibition in the same region during expiration. This finding is consistent with the hypothesis that inhibitory neurons play a crucial role in the inspiration-expiration phase transition in the pBC. To our knowledge this is the first optical imaging of a near fully intact in situ preparation that exhibits both eupneic respiratory activity and functional reflexes.

Action Potentials↗
PubMedDate not supplied

Three-dimensional optical tomographic brain imaging in small animals, part 1: hypercapnia.

In this study, we explore the potential of diffuse optical tomography for brain oximetry. While several groups have already reported on the sensitivity of optical measurements to changes in oxyhemoglobin, deoxyhemoglobin, and blood volume, these studies were often limited to single source-detector geometries or topographic maps, where signals obtained from within the brain are projected onto 2-D surface maps. In this two-part study, we report on our efforts toward developing a volumetric optical imaging system that allows one to spatially resolve 3-D hemodynamic effects in rat brains. In part 1, we describe the instrumentation, optical probe design, and the model-based iterative image reconstruction algorithm employed in this work. Consideration of how a priori anatomical knowledge can be incorporated in the reconstruction process is presented. This system is then used to monitor global hemodynamic changes that occur in the brain under various degrees of hypercapnia. The physiologic cerebral response to hypercapnia is well known and therefore allows an initial performance assessment of the imaging system. As expected, we observe global changes in blood volume and oxygenation, which vary linearly as a function of the concentration of the inspired carbon dioxide. Furthermore, experiments are designed to determine the sensitivity of the reconstructions of only 1 mm to inaccurate probe positioning. We determine that shifts can significantly influence the reconstructions. In part 2 we focus on more local hemodynamic changes that occur during unilateral carotid occlusion performed at lower-than-normal systemic blood pressure. In this case, the occlusion leads to a predominantly monohemispherically localized effect, which is well described in the literature. Having explored the system with a well-characterized physiologic effect, we investigate and discuss the complex compensatory cerebrovascular hemodynamics that occur at normotensive blood pressure. Overall, these studies demonstrate the potential and limitations of our diffuse optical imager for visualizing global and focal hemodynamic phenomenon three dimensionally in the brains of small animals.

Algorithms↗
PubMed2006

Aberrations of chick eyes during normal growth and lens induction of myopia.

Understanding the control of eye growth may lead to the prevention of nearsightedness (myopia). Chicks develop refractive errors in response to defocusing lenses by changing the rate of eye elongation. Changes in optical image quality and the optical signal in lens compensation are not understood. Monochromatic ocular aberrations were measured in 16 chicks that unilaterally developed myopia in response to unilateral goggles with -15D lenses and in 6 chicks developing naturally. There is no significant difference in higher-order root mean square aberrations (RMSA) between control eyes of the goggled birds and eyes of naturally developing chicks. Higher-order RMSA for a constant pupil size exponentially decreases in the chick eye with age more slowly than defocus. In the presence of a defocusing lens, the exponential decrease begins after day 2. In goggled eyes, asymmetric aberrations initially increase significantly, followed by an exponential decrease. Higher-order RMSA is significantly higher in goggled eyes than in controls. Equivalent blur, a new measure of image quality that accounts for increasing pupil size with age, exponentially decreases with age. In goggled eyes, this decrease also occurs after day 2. The fine optical structure, reflected in higher-order aberrations, may be important in understanding normal development and the development of myopia.

Age Factors↗
PubMed2002

Spatial coding of position and orientation in primary visual cortex.

We examined the spatial distribution of population activity in primary visual cortex (V1) of tree shrews with optical imaging and electrophysiology. A line stimulus, thinner than the average V1 receptive field, evoked a broad strip of neural activity of nearly constant size for all stimulus locations tested within the central 10 degrees of visual space. Stimuli in adjacent positions activated highly overlapping populations of neurons; nevertheless, small changes in stimulus position produced orderly changes in the location of the peak of the population response. Statistically significant shifts in the population response were found for stimulus displacements an order of magnitude smaller than receptive field width, down to the limit of optical imaging resolution. Based on the pattern of population activity, we conclude that the map of visual space in V1 is orderly at a fine scale and has uniform coverage of position and orientation without local relationships in the mapping of these features.

Action Potentials↗
PubMed1994

Role of magnesium and calcium in alcohol-induced hypertension and strokes as probed by in vivo television microscopy, digital image microscopy, optical spectroscopy, 31P-NMR, spectroscopy and a unique magnesium ion-selective electrode.

It is not known why alcohol ingestion poses a risk for development of hypertension, stroke and sudden death. Of all drugs, which result in body depletion of magnesium (Mg), alcohol is now known to be the most notorious cause of Mg-wasting. Recent data obtained through the use of biophysical (and noninvasive) technology suggest that alcohol may induce hypertension, stroke, and sudden death via its effects on intracellular free Mg2+ ([Mg2+]i), which in turn alter cellular and subcellular bioenergetics and promote calcium ion (Ca2+) overload. Evidence is reviewed that demonstrates that the dietary intake of Mg modulates the hypertensive actions of alcohol. Experiments with intact rats indicates that chronic ethanol ingestion results in both structural and hemodynamic alterations in the microcirculation, which, in themselves, could account for increased vascular resistance. Chronic ethanol increases the reactivity of intact microvessels to vasoconstrictors and results in decreased reactivity to vasodilators. Chronic ethanol ingestion clearly results in vascular smooth muscle cells that exhibit a progressive increase in exchangeable and cellular Ca2+ concomitant with a progressive reduction in Mg content. Use of 31P-NMR spectroscopy coupled with optical-backscatter reflectance spectroscopy revealed that acute ethanol administration to rats results in dose-dependent deficits in phosphocreatine (PCr), the [PCr]/[ATP] ratio, intracellular pH (pHi), oxyhemoglobin, and the mitochondrial level of oxidized cytochrome oxidase aa3 concomitant with a rise in brain-blood volume and inorganic phosphate. Temporal studies performed in vivo, on the intact brain, indicate that [Mg2+]i is depleted before any of the bioenergetic changes. Pretreatment of animals with Mg2+ prevents ethanol from inducing stroke and prevents all of the adverse bioenergetic changes from taking place. Use of quantitative digital imaging microscopy, and mag-fura-2, on single-cultured canine cerebral vascular smooth muscle, human endothelial, and rat astrocyte cells reveals that alcohol induces rapid concentration-dependent depletion of [Mg2+]i. These cellular deficits in [Mg2+]i seem to precipitate cellular and subcellular disturbances in cytoplasmic and mitochondrial bioenergetic pathways leading to Ca2+ overload and ischemia. A role for ethanol-induced alterations in [Mg2+]i should also be considered in the well-known behavioral actions of alcohol.

Alcoholism↗
PubMed2006

The dynamic range and domain-specific signals of intracellular calcium in photoreceptors.

Vertebrate photoreceptors consist of strictly delimited subcellular domains: the outer segment, ellipsoid, cell body and synaptic terminal, each hosting crucial cellular functions, including phototransduction, oxidative metabolism, gene expression and transmitter release. We used optical imaging to explore the spatiotemporal dynamics of Ca(2+) signaling in non-outer segment regions of rods and cones. Sustained depolarization, designed to emulate photoreceptor activation in the darkness, evoked a standing Ca(2+) gradient in tiger salamander photoreceptors with spatially-averaged intracellular Ca(2+) concentration within synaptic terminals of approximately 2 microM and lower (approximately 750 nM) intracellular calcium concentration in the ellipsoid. Measurements from axotomized cell bodies and isolated ellipsoids showed that Ca(2+) enters the two compartments via both local L-type Ca(2+) channels and diffusion. The results from optical imaging studies were supported by immunostaining analysis. L-type voltage-operated Ca(2+) channels and plasma membrane Ca(2+) ATPases were highly expressed in synaptic terminals with progressively lower expression levels in the cell body and ellipsoid. These results show photoreceptor Ca(2+) homeostasis is controlled in a region-specific manner by direct Ca(2+) entry and diffusion as well as Ca(2+) extrusion. Moreover, quantitative measurement of intracellular calcium concentration levels in different photoreceptor compartments indicates that the dynamic range of Ca(2+) signaling in photoreceptors is approximately 40-fold, from approximately 50 nM in the light to approximately 2 microM in darkness.

Animals↗
PubMed1999

Anatomical correlates of functional plasticity in mouse visual cortex.

Much of what is known about activity-dependent plasticity comes from studies of the primary visual cortex and its inputs in higher mammals, but the molecular bases remain largely unknown. Similar functional plasticity takes place during a critical period in the visual cortex of the mouse, an animal in which genetic experiments can readily be performed to investigate the underlying molecular and cellular events. The experiments of this paper were directed toward understanding whether anatomical changes accompany functional plasticity in the developing visual cortex of the mouse, as they do in higher mammals. In normal mice, transneuronal label after an eye injection clearly delineated the monocular and binocular zones of area 17. Intrinsic signal optical imaging also showed monocular and binocular zones of area 17 but revealed no finer organization of ocular dominance or orientation selectivity. In normal animals, single geniculocortical afferents serving the contralateral eye showed great heterogeneity and no clustering consistent with the presence of ocular dominance patches. Growth and elaboration of terminal arbor continues beyond postnatal day 40 (P40), after the peak of the critical period. After prolonged monocular deprivation (MD) from P20 to P60, transneuronal labeling showed that the projection serving the ipsilateral eye was severely affected, whereas the effect on the contralateral eye's pathway was inconsistent. Optical imaging also showed profound effects of deprivation, particularly in the ipsilateral pathway, and microelectrode studies confirmed continued functional plasticity past P40. Reconstruction of single afferents showed that MD from P20 to P40 promoted the growth of the open eye's geniculocortical connections without causing the closed eye's contralateral projection to shrink, whereas MD from P20 to P60 caused an arrest of growth of deprived arbors. Our findings reveal numerous similarities between mouse and higher mammals in development and plasticity, along with some differences. We discuss the factors that may be responsible for these differences.

Afferent Pathways↗
PubMedDate not supplied

Three-dimensional reconstruction of the cochlea from two-dimensional images of optical sections.

This paper describes a methodology for three-dimensional (3D) computer-aided reconstruction of the guinea pig cochlea using orthogonal-plane fluorescence optical sectioning. Specimens are sectioned optically, allowing them to remain intact during observation. Equations to correct the data for specimen translation and rotation are developed and 3D reconstructions of the scala tympani, round window membrane, and cochlear aqueduct are presented. The error associated with the reconstruction is estimated to be < 19 microns.

Animals↗
PubMed1999

Macroscopic spectral imaging and gene expression analysis of the early stages of melanoma.

BACKGROUND: The stages of melanocytic progression are defined as atypical (dysplastic) nevus, melanoma in situ, melanoma in the radial growth phase (RGP), melanoma in the vertical growth phase (VGP), and melanoma in the metastatic growth phase (MGP). Melanoma in situ and RGP melanoma often develop in contiguous association with atypical nevi. This frequently poses a problem with respect to their early detection. Furthermore, unlike cells obtained from VGP and MGP melanomas, cells derived from melanoma in situ and RGP melanoma do not proliferate in vitro. Thus, compared to the late stages of the disease, less information is available regarding genes expressed in the early stages. MATERIALS AND METHODS: To determine whether spectral imaging, a recently developed optical imaging technique, can detect melanoma in situ and RGP melanoma arising in melanoma precursor lesions, atypical nevi in patients with a clinical history of melanoma were subjected to noninvasive macroscopic spectral imaging. To determine at what stage in the progression pathway of melanoma genes having important biological functions in VGP and MGP melanomas are activated and expressed, lesions of melanoma in situ were analyzed by immunohistochemistry and in situ hybridization for expression of some of these known molecular and immunologic markers. RESULTS: The present study demonstrates the capability of noninvasive spectral imaging to detect melanoma in situ and RGP melanoma that arise in contiguous association with atypical nevi. Furthermore, the study provides evidence that genes and antigens expressed in VGP and MGP melanoma are also expressed in melanoma in situ. CONCLUSIONS: Because of the dark and variegated pigmentation of atypical nevi, melanoma in situ and RGP melanoma that arise in these melanoma precursor lesions are often difficult to recognize and thus frequently go unnoticed. The application of new optical screening techniques for early detection of melanoma and the identification of genes expressed in the early stages of melanoma development are two important avenues in the pursuit of melanoma prevention. The investigations presented here document that macroscopic spectral imaging has the potential to detect melanoma in its early stage of development and that genes essential for the proliferation and cell adhesion of VGP and MGP melanoma are already expressed in melanoma in situ.

Biomarkers, Tumor↗
PubMedDate not supplied

Synthesis, characterization, and biological properties of cyanine-labeled somatostatin analogues as receptor-targeted fluorescent probes.

We present the synthesis and characterization of the somatostatin receptor-specific peptide H(2)N-(D-Phe)-cyclo[Cys-Phe-(D-Trp)-Lys-Thr-Cys]-Thr-OH, which is labeled with a carboxylated indodicarbo- and an indotricarbocyanine dye at the N-terminal amino group. The preparation was performed by automated solid-phase synthesis, with subsequent attachment of the cyanine dye and cleavage of the entire conjugate from the resin. The compounds display high molar absorbance and fluorescence quantum yields typical for cyanine dyes and are thus suitable receptor-targeted contrast agents for molecular optical imaging. The ability of these agents to target the somatostatin receptor was demonstrated by flow cytometry in vitro, in which the indotricarbocyanine conjugate led to elevated cell-associated fluorescence on somatostatin receptor-expressing tumor cells. In contrast, the corresponding linearized derivative of the sequence H(2)N-(D-Phe)-Met-Phe-(D-Trp)-Lys-Thr-Met-Thr-OH produced only minimal cell fluorescence, hence confirming the specificity of the cyclic somatostatin analogue. Intracellular localization could be visualized by near-infrared (NIR) fluorescence microscopy. In conclusion, receptor-specific peptides are promising tools for designing site-directed optical contrast agents for use in molecular optical imaging.

Animals↗
PubMed2001

Building surfaces from borders in Areas 17 and 18 of the cat.

Several brightness illusions indicate that borders can dramatically affect the perception of adjoining surfaces. In the Craik-O'Brien-Cornsweet illusion, in particular, two equiluminant surfaces can appear different in brightness due to the contrast border between them. Although the psychophysical nature of this phenomenon has been well characterized, the neural circuitry underlying this effect is unexplored. Here, we have asked whether there are cells in visual cortex which respond to edge-induced illusory brightness percepts such as the Cornsweet. Using optical imaging and single unit recordings methods, we have studied responses of the primary (Area 17) and second (Area 18) visual cortical areas of the anesthetized cat to both real luminance change and Cornsweet brightness change. We find that there are indeed cells whose responses are modulated in phase with the modulation of the Cornsweet stimulus. These cells are present in both Area 17 and Area 18, but are more prevalent in Area 18. These responses are generally weak and are found even when receptive fields are distant from the contrast border. Consistent with perception, cells which respond to the Cornsweet border are modulated in antiphase to the Narrow Real (another border-induced illusory brightness stimulus). Remarkably, we also find evidence of edge-induced responses to illusory brightness change using intrinsic signal optical imaging. Both real luminance change and edge-induced brightness change produces a greater imaged response in Area 18 than in Area 17. Thus, in the absence of direct luminance stimulation, cells in visual cortex can respond to modulation of distant border contrasts. We suggest that the perception of surface brightness was encoded in the early visual cortical pathway by both surface luminance contrast signals in Area 17 (Rossi, A. F., Rittenhouse, C. D., & Paradiso, M. A. (1996). The representation of brightness in primary visual cortex. Science, 273, 1104-7) and border-induced contrast signals that predominate in Area 18.

Animals↗
PubMed2005

Orientation-restricted continuous visual exposure induces marked reorganization of orientation maps in early life.

To elucidate the effect of visual experience on the development of orientation maps, we conducted intrinsic signal optical imaging of the visual cortex of kittens that were continuously exposed to a single orientation through cylindrical-lens-fitted goggles under a freely moving condition starting at post-natal week 3. We observed a rapid reorganization of orientation maps, characterized by extensive representation of exposed orientations with reduced responsiveness to unexposed orientations. The over-representation of exposed orientation was marked for 1-2 weeks of goggle rearing. A longer period of goggle rearing, however, decreased the degree of over-representation, which still remained at a remarkable level. Dark rearing episodes daily interleaved between single orientation exposures moderated the over-representation effect. Unit recording from goggle-reared kittens showed preferred orientations consistent with optical imaging. Using c-Fos immunoreactivity mapping, we showed that the number of neurons strongly responding to the exposed orientation was 3 times larger in a goggle-reared cat than the number of neurons responding to the vertical orientation in a normal cat. Taken together, these results suggest that the reorganization of orientation maps was caused by the expansion of domains maximally responding to exposed orientation as well as the strong reduction of responses to unexposed orientations.

Algorithms↗
PubMed2004

Molecular expressions: exploring the world of optics and microscopy. http://microscopy.fsu.edu.

Our knowledge of the structure, dynamics and physiology of a cell has increased significantly in the last ten years through the emergence of new optical imaging modalities such as optical sectioning microscopy, computer- enhanced video microscopy and laser-scanning microscopy. These techniques together with the use of genetically engineered fluorophores have helped scientists visualize the 3-dimensional dynamic processes of living cells. However as powerful as these imaging tools are, they can often be difficult to understand and fully utilize. Below I will discuss my favorite website: The Molecular Expressions Web Site that endeavors to present the power of microscopy to its visitors. The Molecular Expressions group does a remarkable job of not only clearly presenting the principles behind these techniques in a manner approachable by lay and scientific audiences alike but also provides representative data from each as well.

Cell Physiological Phenomena↗
PubMed2007

The relation between spike-timing dependent plasticity and Ca2+ dynamics in the hippocampal CA1 network.

In our previous study, spike timing dependent synaptic plasticity (STDP) was investigated in the CA1 area of rat hippocampal slices using optical imaging. It was revealed that the profiles of STDP could be classified into two types depending upon layer specific location along the dendrite. The first was characterized by a symmetric time window observed in the proximal region of the stratum radiatum (SR), and the second by an asymmetric time window in the distal region of the SR. Our methods involved the bath-application of bicuculline (GABA(A) receptor antagonist) to hippocampal slices, which revealed that GABAergic interneuron projections were responsible for the symmetry of a time window. In this study, the intracellular Ca2+ increase of hippocampal CA1 neurons, induced by the protocol of timing between pre- and post-synaptic excitation (i.e. STDP protocol), was measured spatially by using optical imaging to investigate how the triggering of STDP is dependent on intracellular calcium concentration. We found that the magnitude of STDP was closely related to the rate of Ca2+ increase ("velocity") of calcium transient during application of induction stimuli. Location dependency was also analyzed in terms of Ca2+ influx. Furthermore, it was shown that decay time constant of Ca2+ dynamics during the application of STDP-inducing stimuli was also significantly correlated with STDP.

Action Potentials↗
PubMed1998

In vivo fibre optic confocal imaging of microvasculature and nerves in the rat vas deferens and colon.

A fluorescence confocal microscopy technique was employed to obtain subsurface images of nerve and microvascular structure in the vas deferens and colon of the living rat. The use of dual labelling with vital dyes and 2-channel confocal acquisition allowed differentiation of microscopic structure at both low and higher magnification. Characteristic staining patterns of nerves and blood vessels were repeatedly obtained in each tissue, suggesting the potential of this technique for studying morphological changes associated with surgical procedures and/or models of neuronal or vascular pathology.

Animals↗
PubMedDate not supplied

Using near-infrared spectroscopy to assess neural activation during object processing in infants.

The capacity to represent the world in terms of numerically distinct objects (i.e., object individuation) is a milestone in early cognitive development and forms the foundation for more complex thought and behavior. Over the past 10 to 15 yr, infant researchers have expended a great deal of effort to identify the origins and development of this capacity. In contrast, relatively little is known about the neural mechanisms that underlie the ability to individuate objects, in large part because there are a limited number of noninvasive techniques available to measure brain functioning in human infants. Recent research suggests that near-IR spectroscopy (NIRS), an optical imaging technique that uses relative changes in total hemoglobin concentration and oxygenation as an indicator of neural activation, may be a viable procedure for assessing the relation between object processing and brain function in human infants. We examine the extent to which increased neural activation, as measured by NIRS, could be observed in two neural areas known to be involved in object processing, the primary visual cortex and the inferior temporal cortex, during an object processing task. Infants aged 6.5 months are presented with a visual event in which two featurally distinct objects emerge successively to opposite sides of an occluder and neuroimaging data are collected. As predicted, increased neural activation is observed in both the primary visual and inferior cortex during the visual event, suggesting that these neural areas support object processing in the young infant. The outcome has important implications for research in cognitive development, developmental neuroscience, and optical imaging.

Cerebrovascular Circulation↗
PubMed2001

Experimental statistics of near-field intensity distributions at nanostructured surfaces.

Scanning near-field optical microscopy is a technique in which the resolution is primarily determined by the size of a probe and not by the wavelength of illumination as in classical (far-field) microscopy. However, the relationship between a sample and its near-field optical image is usually rather complex. Typical factors responsible, at least partially, for such a complexity are the conditions of illumination and detection, sample characteristics (e.g. roughness and dielectric constant) and optical properties of the probe. Theoretical and experimental works conducted to improve our understanding of the relation between the object and the image have been reported (Greffet & Carminati, 1997). Recently, with the help of a photon scanning tunnelling microscope we have carried out an extensive study of the resultant near-field intensity distributions due to the elastic (in the plane) scattering of surface plasmon polaritons (SPPs) at metal film surfaces. We have also directly observed (in similar experimental conditions) localized dipolar excitations in silver colloid fractals (Bozhevolnyi et al., 1998). In both cases, the studied phenomena are intimately related to the regime of multiple light scattering, in which the interference effects are rather complicated and therefore a proper interpretation of them was far from being trivial. Thus, even though a certain understanding of many features inherent to the subwavelength light interference phenomena was gained (Bozhevolnyi & Coello, 1998; Bozhevolnyi et al., 1998; Coello & Bozhevolnyi, 1999), it is clear from the outcome of the investigations that more systematic studies in this context are still needed. A different and more powerful approach may be a statistical study of the recorded near-field intensity distributions. In this work, we report what we believe to be the first results on experimental statistics of near-field optical images exhibiting localized optical excitations (related to the regime of multiple scattering of light). We investigated optical images obtained with SPPs excited at different light wavelengths and scattered at different film surfaces, and with different polarizations and wavelengths of light scattered by silver colloid fractal structures. We have found significant differences in statistics between near-field intensity distributions taken at rough and smooth metal film surfaces and fractal structures. Finally, our predictions seem to be in agreement with theoretical studies reported by other authors (Sanchez-Gil & Garcia-Ramos, 1998).

Journal Article↗
PubMed2002

Optical-based molecular imaging: contrast agents and potential medical applications.

Laser- and sensitive charge-coupled device technology together with advanced mathematical modelling of photon propagation in tissue has prompted the development of novel optical imaging technologies. Fast surface-weighted imaging modalities, such as fluorescence reflectance imaging (FRI) and 3D quantitative fluorescence-mediated tomography have now become available [1, 2]. These technical advances are paralleled by a rapid development of a whole range of new optical contrasting strategies, which are designed to generate molecular contrast within a living organism. The combination of both, technical advances of light detection and the refinement of optical contrast media, finally yields a new spectrum of tools for in vivo molecular diagnostics. Whereas the technical aspects of optical imaging are covered in more detail in a previous review article in "European Radiology" [3], this article focuses on new developments in optical contrasting strategies and design of optical contrast agents for in vivo diagnostics.

Animals↗