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Results for “Optical imaging”

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At least 505 records · Page 28Linked to original sources

Nanoscale optical imaging of excitons in single-walled carbon nanotubes.

We present simultaneous near-field photoluminescence (PL) and Raman imaging of single-walled carbon nanotubes (SWNTs) with a spatial resolution better than 15 nm. Highly localized excitation is used to visualize the spatial extent of the contributing excited states. For SWNTs on glass, we typically observe highly confined PL from short segments of about 20 nm in length. The PL from micelle-encapsulated SWNTs on mica is extended along the tube up to several hundreds of nanometers. We find that near-field enhancement is much stronger for photoluminescence than for Raman scattering, an observation that is explained by the low intrinsic quantum yield of SWNTs.

Journal Article↗

Optical imaging of calcium accumulation in hippocampal pyramidal cells during synaptic activation.

The dynamic response of nerve cells to synaptic activation and the spatial distribution of biochemical processes regulated by ion concentration are critically dependent on the cell-surface distribution of ion channels. In the hippocampus, intracellular calcium-ion concentration is thought to influence the biochemical events associated with kindling, excitotoxicity, and long-term potentiation. Computer models of hippocampal pyramidal cells also indicate that calcium-channel location influences dynamic characteristics such as bursting. Here, we have used in situ microfluorometric imaging in brain slices to directly measure the spatial distribution of calcium accumulation in guinea-pig CA1 pyramidal cells during trains of orthodromic synaptic stimulation. Calcium accumulation is substantial throughout the entire proximal section of the apical and basal dendrites. Most of this accumulation results from influx through non-NMDA (N-methyl-D-aspartate) voltage-gated calcium channels, and in the apical dendrite it drops steeply as the dendrite enters stratum moleculare, the termination zone of perforant path afferents. These results demonstrate a marked segregation of calcium-channel activity and directly show a spatial distribution of calcium accumulation during orthodromic synaptic activation.

Animals↗

Visualization of advanced human prostate cancer lesions in living mice by a targeted gene transfer vector and optical imaging.

Non-invasive imaging and transcriptional targeting can improve the safety of therapeutic approaches in cancer. Here we demonstrate the ability to identify metastases in a human-prostate cancer model, employing a prostate-specific adenovirus vector (AdPSE-BC-luc) and a charge-coupled device-imaging system. AdPSE-BC-luc, which expresses firefly luciferase from an enhanced prostate-specific antigen promoter, restricted expression in the liver but produced robust signals in prostate tumors. In fact, expression was higher in advanced, androgen-independent tumors than in androgen-dependent lesions. Repetitive imaging over a three-week period after AdPSE-BC-luc injection into tumor-bearing mice revealed that the virus could locate and illuminate metastases in the lung and spine. Systemic injection of low doses of AdPSE-BC-luc illuminated lung metastasis. These results demonstrate the potential use of a non-invasive imaging modality in therapeutic and diagnostic strategies to manage prostate cancer.

Animals↗

Herpesvirus saimiri-based vector biodistribution using noninvasive optical imaging.

Herpesvirus saimiri (HVS) is capable of infecting a range of human cell types with high efficiency and the viral genome persists as high copy number, circular, nonintegrated episomes which segregate to progeny upon cell division. This allows the HVS-based vector to stably transduce a dividing cell population and provide sustained transgene expression for an extended period of time both in vitro and in vivo. Here we assess the dissemination of HVS-based vectors in vivo following intravenous and intraperitoneal administration. Bioluminescence imaging of an HVS-based vector expressing luciferase demonstrates that the virus can infect and establish a persistent latent infection in a variety of mouse tissues. Moreover, the long-term in vivo maintenance of the HVS genome as a nonintegrated circular episome provided sustained expression of luciferase over a 10-week period. A particularly high level of transgene expression in the liver and the ability of HVS to infect and persist in hepatic stellate cells suggest that HVS-based vectors may have potential for the treatment of inherited and acquired liver diseases.

Animals↗

Optical imaging of hippocampal neurons with a chloride-sensitive dye: early effects of in vitro ischemia.

We determined if changes in intraneuronal Cl- occur early after ischemia in the hippocampal slice. Slices from juvenile rats (14-19 days old) were loaded with the cell-permeant form of 6-methoxy-N-ethylquinolinium chloride (MEQ), a Cl(-)-sensitive fluorescent dye. Real-time changes in intracellular chloride concentration ([Cl-]i) were measured with UV laser scanning confocal microscopy in multiple neurons within each slice. In vitro ischemia (26-28 degrees C, 10 min) was confirmed by the loss of synaptic transmission (evoked field excitatory postsynaptic potentials) from pyramidal cells in area CA1. After ischemia and reoxygenation (10 min), MEQ fluorescence decreased significantly in CA1 pyramidal cells and interneurons. The decreased fluorescence corresponded to an ischemia-induced increase in [Cl-]i of approximately 10 mM. Pretreatment with the GABA(A)-gated Cl- channel antagonist picrotoxin (100 microM) blocked the ischemia-induced change in [Cl-]i. Analysis of the superfusates indicated that ischemia also caused a transient amino acid (GABA, glutamate, and aspartate) release that was maximal at approximately 10 min, returning to baseline shortly thereafter. Recovery from ischemia was confirmed by the return of synaptic transmission in area CA1, the return toward baseline of the ischemia-induced decrease in MEQ fluorescence, and exclusion of propidium iodide from MEQ fluorescent cells. Furthermore, pyramidal cells did not undergo cell swelling during this early phase of reoxygenation, as indicated by the volume-sensitive dye calcein. Thus, mild ischemia induces the accumulation of [Cl-]i secondary to GABA(A) receptor activation, in the absence of cellular swelling or death. In contrast, depolarization of the slice with K+ (50 mM) decreased MEQ fluorescence significantly but caused cell swelling. Picrotoxin did not prevent the K+-induced increase in [Cl-]i. It is possible that an increased [Cl-]i, following either an ischemic event or an episode of depolarization, would reduce the Cl- driving force and thereby limit synaptic transmission by GABA. To support this hypothesis, ischemia caused a reduction in the ability of the GABA agonist muscimol to increase [Cl-]i after 20-min reoxygenation.

Animals↗

Optical imaging of Renilla luciferase reporter gene expression in living mice.

Imaging reporter gene expression in living subjects is a rapidly evolving area of molecular imaging research. Studies have validated the use of reporter genes with positron emission tomography (PET), single photon emission computed tomography (SPECT), MRI, fluorescence with wild-type and mutants of green fluorescent protein, as well as bioluminescence using Firefly luciferase enzyme/protein (FL). In the current study, we validate for the first time the ability to image bioluminescence from Renilla luciferase enzyme/protein (RL) by injecting the substrate coelenterazine in living mice. A highly sensitive cooled charge-coupled device camera provides images within a few minutes of photon counting. Cells, transiently expressing the Rluc were imaged while located in the peritoneum, s.c. layer, as well as in the liver and lungs of living mice tail-vein injected with coelenterazine. Furthermore, d-luciferin (a substrate for FL) does not serve as a substrate for RL, and coelenterazine does not serve as a substrate for FL either in cell culture or in living mice. We also show that both Rluc and Fluc expression can be imaged in the same living mouse and that the kinetics of light production are distinct. The approaches validated will have direct applications to various studies where two molecular events need to be tracked, including cell trafficking of two cell populations, two gene therapy vectors, and indirect monitoring of two endogenous genes through the use of two reporter genes.

Animals↗

Optical image quality in the peripheral retina.

A double-pass, photoelectric, ophthalmoscopic method was used for objective study of the variation in image quality across the retina in terms of the reflected image of a fine line, the line-spread function. Radial and tangential line-spread functions have been studied with active and paralyzed accommodation at eccentricities up to 45 deg. With large pupils, the retinal image quality remained relatively constant over the central 20 deg of field and deteriorated at larger field angles. The results are discusssed with respect to field curvature, peripheral acuity, and peripheral refraction.

Adult↗

Scatter and its implications for the measurement of optical image quality in human eyes.

PURPOSE: To investigate the effect of scatter on measurements of wavefront aberrations and point-spread functions in a model eye. METHODS: The wavefront aberrations of a model eye were measured using Hartmann-Shack wavefront sensing and crossed-cylinder aberroscope techniques and compared with its measured point-spread function in the presence of scattering media of different concentrations. RESULTS: The point-spread functions became broader as the concentration increased. Forward light scatter on both the light path into the eye and the light path out of the eye contributed to this broadening of the point-spread function. Neither the crossed-cylinder aberroscope nor wavefront sensing, which, respectively, measure the ocular wavefront aberrations for light entering the eye and leaving the eye, were affected by the scatter. CONCLUSION: We predict that by minimizing the contribution of the forward light scatter from one or other of these light paths by manipulating the size of the entrance and exit pupils, it should be possible to objectively assess narrow-angle forward light scatter in the eye by measuring and removing any confounding effect from wavefront aberration.

Diagnostic Techniques, Ophthalmological↗

Surpassing the standard quantum limit for optical imaging using nonclassical multimode light.

Using continuous wave superposition of spatial modes, we demonstrate experimentally displacement measurement of a light beam below the standard quantum limit. Multimode squeezed light is obtained by mixing a vacuum squeezed beam and a coherent beam that are spatially orthogonal. Although the resultant beam is not squeezed, it is shown to have strong internal spatial correlations. We show that the position of such a light beam can be measured using a split detector with an increased precision compared to a classical beam. This method can be used to improve the sensitivity of small displacement measurements.

Journal Article↗

Time-resolved reflectance at null source-detector separation: improving contrast and resolution in diffuse optical imaging.

We propose a novel approach to imaging in diffusive media based on time-resolved reflectance measurements at null source-detector separation. This approach yields better spatial resolution and contrast as compared to the classical approach, which typically employs a separation of 20-40 mm. Results are obtained by an analytical perturbation approach to diffusion theory and on Monte Carlo simulations. Practical implementation with state-of-the-art technology and performance of a complementary approach based on the use of small but not null source-detector separation are also discussed.

Journal Article↗

Subwavelength optical imaging through a metallic nanorod array.

We propose a subwavelength imaging system without a lens or a mirror but with an array of metallic nanorods. The near-field components of dipole sources were plasmonically transferred through the rod array to reproduce the source distribution in the other side. We calculated the field distribution at the different planes of imaging process using the finite-difference time-domain algorithm and found that the spatial resolution was 40 nm given by the rod size and spacing. A typical configuration is a hexagonal arrangement of silver rods of 50 nm height and 20 nm diameter. We also show that the image formation highly depends on the coherence and the polarization of the source distribution and the source-array distance.

Journal Article↗

Functional near-infrared optical imaging: utility and limitations in human brain mapping.

Although near-infrared spectroscopy (NIRS) was developed as a tool for clinical monitoring of tissue oxygenation, it also has potential for neuroimaging. A wide range of different NIRS instruments have been developed, and instruments for continuous intensity measurements with fixed spacing [continuous wave (CW)-type instruments], which are most readily available commercially, allow us to see dynamic changes in regional cerebral blood flow in real time. However, quantification, which is necessary for imaging of brain functions, is impossible with these CW-type instruments. Over the past 20 years, many different approaches to quantification have been tried, and several multichannel time-resolved and frequency-domain instruments are now in common use for imaging. Although there are still many problems with this technique, such as incomplete knowledge of how light propagates through the head, NIRS will not only open a window on brain physiology for subjects who have rarely been examined until now, but also provide a new direction for functional mapping studies.

Algorithms↗