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At least 1,081 records · Page 60Linked to original sources
PubMed2006

Normalized transillumination of fluorescent proteins in small animals.

We investigated the capacity of normalized transillumination methods in imaging fluorescent proteins in visible light, in phantoms and in live mice. We demonstrate significant imaging improvements over epi-illumination imaging, as commonly applied today. Of particular importance is the significant betterment in contrast achieved, due to minimization of the influence of autofluorescence and the enhancement of depth sensitivity compared to epi-illumination imaging. Although normalized transillumination does not match the accuracy and quantification capacity of tomographic imaging, it nevertheless attains key advantages over other planar optical imaging methods while preserving implementation simplicity. Due to the balance achieved between simplicity and accuracy, normalized transillumination approaches could serve as an important alternative molecular imaging method.

Animals↗
PubMed2006

Benefiting from the unique properties of lanthanide ions.

The recent upsurge of interest in contrast agents for magnetic resonance imaging, of luminescent chemosensors for medical diagnostic, and lately, for optical imaging of cells has generated an impressive momentum for the coordination and supramolecular chemistry of trivalent lanthanide ions. We shortly review the synthetic methods allowing the introduction of these spherical ions with fascinating optical and magnetic properties into elaborate mono- and polymetallic edifices. We then illustrate these methods by selected examples describing the use of (i) a coronand to produce luminescent liquid crystals, (ii) derivatized calixarenes for 4f-5f element separation, (iii) podates for the production of nanoparticles with high relaxivity and for sensitizing the near-infrared (NIR) emission, and (iv) self-assembly processes for producing functional bimetallic edifices.

Ions↗
PubMedDate not supplied

Contrast-detail analysis characterizing diffuse optical fluorescence tomography image reconstruction.

Contrast-detail analysis is used to evaluate the imaging performance of diffuse optical fluorescence tomography (DOFT), characterizing spatial resolution limits, signal-to-noise limits, and the trade-off between object contrast and size. Reconstructed images of fluorescence yield from simulated noisy data were used to determine the contrast-to-noise ratio (CNR). A threshold of CNR=3 was used to approximate a lowest acceptable noise level in the image, as a surrogate measure for human detection of objects. For objects 0.5 cm inside the edge of a simulated tissue region, the smallest diameter that met this criteria was approximately 1.7 mm, regardless of contrast level, and test field diameter had little impact on this limit. Object depth had substantial impact on object CNR, leading to a limit of 4 mm for objects near the center of a 51-mm test field and 8.5 mm for an 86-mm test field. Similarly, large objects near the edge of both test fields required a minimum contrast of 50% to achieve acceptable image CNR. The minimum contrast for large, centered objects ranged between 50% and 100%. Contrast-detail analysis using human detection of lower contrast limits provides fundamentally important information about the performance of reconstruction algorithms, and can be used to compare imaging performance of different systems.

Algorithms↗
PubMed2002

Voltage-sensitive dye imaging of population neuronal activity in cortical tissue.

Voltage-sensitive dyes (VSDs) and optical imaging are useful for studying spatiotemporal patterns of population neuronal activity in cortical tissue. Using a photodiode array and absorption dyes we were able to detect neuronal activity in single trials before it could be detected by local field potential (LFP) recordings. Simultaneous electrical and optical recordings from the same tissue also showed that VSD and LFP signals have different waveforms during different activities, suggesting that they are sensitive to different aspects of the synchronization across the population. Noise, dye bleaching, phototoxicity and optical filter selection are important to the quality of the VSD signal and are discussed in this report. With optimized signal-to-noise ratio (S/N) and total recording time, we can optically monitor approximately 500 locations in an area of 1 mm(2) of cortical tissue with a sensitivity comparable to that of LFP electrodes. The total recording time and S/N of fluorescence and absorption dyes are also compared. At S/N of 8-10, absorption dye NK3630 allows a total recording time of 15-30 min, which can be divided into hundreds of 4-8 s recording trials over several hours, long enough for many kinds of experiments. In conclusion, the VSD method provides a reliable way for examining neuronal activity and pharmacological properties of synapses in brain slices.

Action Potentials↗
PubMed1988

Phase-transfer function of the human eye and its influence on point-spread function and wave aberration.

The bidimensional phase-transfer function (PTF) of the human eye has been computed from aerial retinal images of a point test. These images were previously determined by using a recently developed hybrid optical-digital method. Actual PTF data have been obtained directly without linear variations with spatial frequency and have shown great variations among individual subjects. The influence of the PTF on the determination of the point-spread function and the wave-aberration function for emmetropized and slightly astigmatic subjects has been also evaluated. Finally, the effect of pupil size on the PTF was determined by computing these functions from the wave aberration. These results allow us to give a more thorough description of the optical image quality of the human eye and can be used as actual data in subsequent psychophysical studies.

Humans↗
PubMedDate not supplied

Image analysis methods for diffuse optical tomography.

Three major analytical tools in imaging science are summarized and demonstrated relative to optical imaging in vivo. Standard resolution testing is optimal when infinite contrast is used and hardware evaluation is the goal. However, deep tissue imaging of absorption or fluorescent contrast agents in vivo often presents a different problem, which requires contrast-detail analysis. This analysis shows that the minimum detectable sizes are in the range of 1/10 the outer diameter, whereas minimum detectable contrast values are in the range of 10 to 20% relative to the continuous background values. This is estimated for objects being in the center of the domain being imaged, and as the heterogeneous region becomes closer to the surface, the lower limit on size and contrast can become arbitrarily low and more dictated by hardware specifications. Finally, if human observer detection of abnormalities in the images is the goal, as is standard in most radiological practice, receiver operating characteristic (ROC) curve and location receiver operating characteristic curve (LROC) are used. Each of these three major areas of image interpretation and analysis are reviewed in the context of medical imaging as well as how they are used to quantify the performance of diffuse optical imaging of tissue.

Algorithms↗
PubMed1997

Assessment of dental caries with Digital Imaging Fiber-Optic TransIllumination (DIFOTI): in vitro study.

This paper describes Digital Imaging Fiber-Optic TransIllumination (DIFOTI-TM), a new method for the reliable detection of dental caries. Images of teeth obtained through visible-light, fiber-optic transillumination (FOTI) are acquired with a digital CCD camera, and sent to a computer for analysis with dedicated algorithms. The algorithms were developed to facilitate the location and diagnosis of carious lesions by the operator in real time, and provide quantitative characterization for monitoring of the lesions. The DIFOTI method has been tested by imaging teeth in vitro. The results suggest the superior sensitivity of DIFOTI for detection of approximal, occlusal and smooth-surface caries vis-à-vis radiological imaging.

Dental Caries↗
PubMed2001

Voltage-sensitive dye imaging of neocortical spatiotemporal dynamics to afferent activation frequency.

The spatial and temporal patterns of neocortex activation are determined not only by the dynamic character of the input but also by the intrinsic dynamics of the cortical circuitry. To study the role of afferent input frequency on cortical activation dynamics, the electrical activity of in vitro neocortex slices was imaged during white-matter electrical stimulation. High-speed optical imaging was implemented using voltage-sensitive dyes in guinea pig visual and somatosensory cortex slices concomitantly with intracellular recordings. Single white-matter electrical stimuli activated well-defined cortical sites with a radially oriented columnar configuration. This configuration was followed, over the next few milliseconds, by a lateral spread of excitation through cortical layers 5 and 6 and layers 2 and 3. Much of the optical response was eliminated in low extracellular calcium, indicating that it was primarily synaptically mediated. Repetitive stimuli at 10 Hz reproduced the spatiotemporal pattern observed for single stimuli. In contrast, repetitive stimulation in the gamma frequency range ( approximately 40 Hz) rapidly restrained the area of excitation to a small columnar site directly above the stimulating electrode. Intracellular recordings from cells lateral to the activated column revealed increased inhibitory synaptic activity and/or decreased excitatory responses during the train at 40 Hz, but not during a 10 Hz stimulation. Localized microinjections of GABA(A) antagonist produced a reorganization of the geometrical activity pattern that was dependent on the position of the microinjection site. These findings indicate that the frequency-dependent spatial organization of neocortex activation is determined by inhibitory sculpting attributable to local network dynamics.

Afferent Pathways↗
PubMedDate not supplied

Use-dependent plasticity in barrel cortex: intrinsic signal imaging reveals functional expansion of spared whisker representation into adjacent deprived columns.

We used optical imaging of intrinsic cortical signals, elicited by whisker stimulation, to define areas of activation in primary sensory cortex of normal hamsters and hamsters subjected to neonatal follicle ablation at postnatal day seven (P7). Follicle ablations were unilateral, and spared either C-row whiskers or the second whisker arc. This study was done to determine if the intrinsic cortical connectivity pattern of the barrel cortex, established during the critical period, affects the process of representational plasticity that follows whisker follicle ablation. Additionally, we tested the ability to monitor such changes in individual cortical whisker representations using intrinsic signal imaging. Stimulation of a single whisker yielded peak activation of a barrel-sized patch in the somatotopically appropriate location in normal cortex. In both row and arc-spared animals, functional representations corresponding to spared follicles were significantly stronger and more oblong than normal. The pattern of activation differed in the row-sparing and arc-sparing groups, in that the expansion was preferentially into deprived, not spared areas. Single whisker stimulation in row-spared cases preferentially activated the corresponding barrel arc, while stimulation of one whisker in arc-spared cases produced elongated activation down the barrel row. Since whisker deflection normally has a net inhibitory effect on neighboring barrels, our data suggest that intracortical inhibition fails to develop normally in deprived cortical columns. Because thalamocortical projections are not affected by follicle ablation after P7, we suggest that the effects we observed are largely cortical, not thalamocortical.

Animals↗
PubMed2003

Metabolism-enhanced tumor localization by fluorescence imaging: in vivo animal studies.

We present a high-sensitivity near-infrared optical imaging system for noninvasive cancer detection and localization based on molecularly labeled fluorescent contrast agents. This frequency-domain system utilizes the interferencelike pattern of diffuse photon density waves to achieve high detection sensitivity and localization accuracy for the fluorescent heterogeneity embedded inside the scattering media. A two-dimensional localization map is obtained through reflectance probe geometry and goniometric reconstruction. In vivo measurements with a tumor-bearing mouse model by use of the novel Cypate-mono-2-deoxy-glucose fluorescent contrast agent, which targets the enhanced tumor glycolysis, demonstrate the feasibility of detection of a 2-cm-deep subsurface tumor in the tissuelike medium, with a localization accuracy within 2-3 mm.

Animals↗
PubMed2003

Time-resolved luminescence imaging of hydrogen peroxide using sensor membranes in a microwell format.

We demonstrate an optical imaging scheme for hydrogen peroxide in a microwell-based format using the europium(III) tetracycline complex as the fluorescent probe, which is incorporated into a polyacrylonitrile-co-polyacrylamide polymer matrix. The resulting sensor membranes are integrated into a 96-microwell plate. Hydrogen peroxide can be visualized by means of time-resolved luminescence lifetime imaging. The imaging system consists of a fast, gated charge-coupled device (CCD) camera and a pulsed array of 96 light emitting diodes (LEDs). Fluorescence lifetime images are acquired in different modes (rapid lifetime determination, RLD, and phase delay rationing, PDR) and compared with conventional intensity-based methods with respect to sensitivity and the dynamic range of the sensor. The lowest limits of detection can be achieved by the RLD method. The response time of the sensor is comparatively high, typically in the range of 10 to 20 minutes, but the response is reversible. The largest signal changes are observed at pH values between 6.5 and 7.5.

Adsorption↗
PubMed2006

New field with tonotopic organization in guinea pig auditory cortex.

In guinea pig auditory cortex, two core areas, a primary area (AI) and a dorsocaudal field (DC), and two belt regions ventral to AI and DC (VRB and VCB) with an intermediate zone (T) in between, together with a small field (S) rostral to AI, have been reported in single-electrode studies although field S and zone T have not been observed in imaging studies. Using a high-resolution in vivo optical-imaging system with the voltage-sensitive dye RH-795, we report here the successful imaging of a rostral small field and zone T and a ventral-to-dorsal frequency gradient in zone T. Further, we found that VRB can be subdivided into two areas, a ventrorostral field (VR) with properties similar to those reported for VRB, and a ventrocaudal field (VC) with novel properties. With increasing stimulus tone frequency, activation in VR shifted caudally while activation in VC shifted rostrally. Thus we have newly identified field VC that has mirror-symmetric tonotopy to that of VR.

Acoustic Stimulation↗
PubMed1993

NMR and time-resolved optical studies of brain imaging.

The classical optical method of the biochemist used in so many applications to determine stoichiometry kinetics, quantitative analysis of biochemical systems which was subsequently applied to tissue studies by G.A. Millikan and others seems now to have a new lease on life provided by time and frequency resolved studies which appear to afford the essential quantification lacking in the earlier methods. The fact that localization seems possible as well may bring the optical technology into the arena of new non-invasive technology capable of high sensitivity, high resolution and rapid data acquisition.

Animals↗
PubMed1987

STIR sequences in NMR imaging of the optic nerve.

Orbital fat surrounding the optic nerve causes considerable difficulties in NMR imaging due to its high image intensity and the chemical shift artefact. We have investigated the ability of inversion recovery sequences with short inversion times (STIR sequences) to suppress fat signals in imaging the optic nerve. We have also compared the contrast attainable with STIR sequences with that obtainable from other sequences. Measurements were made on 4 normal controls and 5 patients with multiple sclerosis (MS) to obtain typical values of relaxation times and proton densities for orbital fat, cerebral white matter and MS lesions. The fat T1 measurements were used to predict an appropriate inversion time for the STIR sequence and estimate how much residual fat signal might be expected as a result of natural variations in fat T1. STIR sequences can be used to suppress the signal from orbital fat with little residual signal. Measurements from white matter and MS lesions were used to predict the contrast between normal and pathological tissues that is attainable with STIR sequences. STIR contrast compares favourably with that obtainable from other sequences.

Humans↗
PubMed2001

Tuning and topography in an odor map on the rat olfactory bulb.

The sense of smell originates in a diverse array of receptor neurons, comprising up to 1000 different types. To understand how these parallel channels encode chemical stimuli, we recorded the responses of glomeruli in the olfactory bulbs of the anesthetized rat, by optical imaging of intrinsic signals. Odor stimulation produced two kinds of optical responses at the surface of the bulb: a broad diffuse component superposed by discrete small spots. Histology showed that the spots correspond to individual glomeruli, and that approximately 400 of them can be monitored in this way. Based on its wavelength-dependence, this optical signal appears to derive from changes in light scattering during neural activity. Pure odorants generally activated several glomeruli in a bilaterally symmetric pattern, whose extent varied greatly with concentration. A simple formalism for ligand binding accounts quantitatively for this concentration dependence and yields the effective affinity with which a glomerulus responds to an odorant. When tested with aliphatic molecules of increasing carbon chain length, many glomeruli were sharply tuned for one or two adjacent chain lengths. Glomeruli with similar tuning properties were located near each other, producing a systematic map of molecular chain length on the surface of the olfactory bulb. Given local inhibitory circuits within the olfactory bulb, this can account for the observed functional inhibition between related odors. We explore several parallels to the function and architecture of the visual system that help interpret the neural representation of odors.

Administration, Inhalation↗
PubMed1996

Quantitative long-term imaging of the functional representation of a whisker in rat barrel cortex.

In this study, we implement chronic optical imaging of intrinsic signals in rat barrel cortex and repeatedly quantify the functional representation of a single whisker over time. The success of chronic imaging for more than 1 month enabled an evaluation of the normal dynamic range of this sensory representation. In individual animals for a period of several weeks, we found that: (i) the average spatial extent of the quantified functional representation of whisker C2 is surprisingly large--1.71 mm2 (area at half-height); (ii) the location of the functional representation is consistent; and (iii) there are ongoing but nonsystematic changes in spatiotemporal characteristics such as the size, shape, and response amplitude of the functional representation. These results support a modified description of the functional organization of barrel cortex, where although a precisely located module corresponds to a specific whisker, this module is dynamic, large, and overlaps considerably with the modules of many other whiskers.

Animals↗
PubMed2001

Depth dependence of the analytical expression for the width of the point spread function (spatial resolution) in time-resolved transillumination.

Simple analytical expressions for the point spread function (PSF) at different depths can save computation time and improve the performance of inverse algorithms for optical imaging. In particular, application of such formulas simplifies quantification of the optical characteristics of tissue abnormalities inside highly scattering media. Earlier it was shown within the random walk theory framework that the PSF for time-resolved transillumination imaging through a highly scattering slab is well represented by a Gaussian. We have experimentally validated a simple formula of the random walk model for the effective width of this Gaussian, as a function of time delay, at different depths of the target. Presented analysis of published experimental data, concerning effective width of the PSF, for a slab of considerably smaller thickness also demonstrates good agreement between the data and predictions of our model. This PSF width determines spatial resolution of the time-resolved imaging and is widely discussed in the literature.

Diagnostic Imaging↗
PubMed1997

Fast temporal response fiber-optic chemical sensors based on the photodeposition of micrometer-scale polymer arrays.

Fiber-optic chemical sensor microarrays for the detection of pH and O2 have been developed with subsecond response times. Sensor microarrays are fabricated by the covalent immobilization (pH sensor arrays) or the physical entrapment (O2 sensor arrays) of fluorescent indicators in photodeposited polymer matrices on optical imaging fibers. Polymer microarrays are comprised of thousands of individual elements photodeposited as hemispheres such that each element of the sensor array is coupled directly to a discrete optical element of the imaging fiber and is not in contact with other neighboring elements. Because of the hemispherical shape and the individuality of the array elements, diffusion of analyte to the sensor elements is dominated by radial diffusion, resulting in a rapid response time. pH-sensitive arrays based on fluorescein respond to a 1.5-unit pH change within 300 ms, while the O2-sensitive arrays respond to O2 changes within 200 ms (90% of steady state response).

Biosensing Techniques↗