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PubMedDate not supplied

Direct digitization of optical images using a photostimulable phosphor system.

The authors describe a method for directly digitizing optical images with a photostimulable phosphor (PSP) system. A PSP plate is initially charged with an exposure to a uniform x-ray field, and is then exposed to an optical image which discharges the plate in relation to the amount of incident light. Two applications were investigated: a contact-print technique for digitizing film radiographs, and a projection technique for digitizing transparent objects such as histology slides. Spatial uniformity was found to be adequate, and linearity of optical density response was excellent from 0.0-2.9 o.d. after look-up table correction. Spatial frequency response was degraded with the optical technique relative to the x-ray imaging properties of the plates, but was restorable by Fourier filtering. Image noise following spatial enhancement was satisfactory at intermediate to high optical densities using a high-resolution PSP plate, but was somewhat degraded at low densities.

Humans↗
PubMed2003

[Accurate establishment of the retinotopic topography of area 17 in cats by intrinsic signal optical imaging].

The retinotopic topography of area 17 in cats was measured by optical imaging based on intrinsic signals. When stimulated with two neighboring gratings oriented orthogonally each other, which were positioned respectively in the upper and lower visual fields, one piece of cortex that had the retinal projection corresponding to the area around the border of the two stimulus gratings became blurred in the resultant function orientation map, because the neurons in this site received excitatory signals from both the horizontal and the vertical gratings via indirect ways. This functional map of the same cortex was compared with that elicited only by a horizontal or vertical grating stimulation in the whole visual field. Accordingly, the accurate position of the retinotopic eccentricity of the cortex in visual field can be demarcated by calculating the cross correlation coefficient of the two functional maps. Furthermore, compared with the electrophysiological measure of receptive fields of single cortical neurons, the retinotopic eccentricities revealed by optical imaging were identical. This experiment provides a fast and relatively accurate method to calculate the retinotopic eccentricities in a large cortical area of the visual cortex.

Animals↗
PubMed2004

Diffuse optical imaging of brain activation: approaches to optimizing image sensitivity, resolution, and accuracy.

Near-infrared spectroscopy (NIRS) and diffuse optical imaging (DOI) are finding widespread application in the study of human brain activation, motivating further application-specific development of the technology. NIRS and DOI offer the potential to quantify changes in deoxyhemoglobin (HbR) and total hemoglobin (HbT) concentration, thus enabling distinction of oxygen consumption and blood flow changes during brain activation. While the techniques implemented presently provide important results for cognition and the neurosciences through their relative measures of HbR and HbT concentrations, there is much to be done to improve sensitivity, accuracy, and resolution. In this paper, we review the advances currently being made and issues to consider for improving optical image quality. These include the optimal selection of wavelengths to minimize random and systematic error propagation in the calculation of the hemoglobin concentrations, the filtering of systemic physiological signal clutter to improve sensitivity to the hemodynamic response to brain activation, the implementation of overlapping measurements to improve image spatial resolution and uniformity, and the utilization of spatial prior information from structural and functional MRI to reduce DOI partial volume error and improve image quantitative accuracy.

Algorithms↗
PubMed2002

Optical imaging reveals retinotopic organization of dorsal V3 in New World owl monkeys.

Optical imaging of intrinsic responses to visual stimuli in extrastriate cortex of owl monkeys provided evidence for the dorsal half of the third visual area, V3. Visual stimuli were used to selectively activate locations in dorsolateral V2 and the rostrally adjoining presumptive V3. Consistent with the proposed retinotopies of dorsal V2 and dorsal V3, small bars of drifting gratings along the horizontal meridian of the contralateral hemifield activated cortex along the V2V3 border, whereas such stimuli along the vertical meridian activated cortex along the rostral border of V3. Stimuli in limited locations in the lower visual quadrant revealed mirror reversals of retinotopy in dorsal V2 and V3, whereas stimuli in the upper visual quadrant failed to activate either region. Brain sections processed for cytochrome oxidase from the same cases provided architectonic borders of V2 that matched those indicated by the optical imaging. The results support the concept that a narrow dorsal V3 exists in monkeys. V3d borders dorsal V2 and contains a smaller, mirror-image representation of the lower visual quadrant.

Action Potentials↗
PubMed2001

Novel fluorescent contrast agents for optical imaging of in vivo tumors based on a receptor-targeted dye-peptide conjugate platform.

We have designed, synthesized, and evaluated the efficacy of novel dye-peptide conjugates that are receptor specific. Contrary to the traditional approach of conjugating dyes to large proteins and antibodies, we used small peptide-dye conjugates that target over-expressed receptors on tumors. Despite the fact that the peptide and the dye probe have similar molecular mass, our results demonstrate that the affinity of the peptide for its receptor and the dye fluorescence properties are both retained. The use of small peptides has several advantages over large biomolecules, including ease of synthesis of a variety of compounds for potential combinatorial screening of new targets, reproducibility of high purity compounds, diffusiveness to solid tumors, and the ability to incorporate a variety of functional groups that modify the pharmacokinetics of the peptide-dye conjugates. The efficacy of these new fluorescent optical contrast agents was evaluated in vivo in well-characterized rat tumor lines expressing somatostatin (sst(2)) and bombesin receptors. A simple continuous wave optical imaging system was employed. The resulting optical images clearly show that successful specific tumor targeting was achieved. Thus, we have demonstrated that small peptide-dye conjugates are effective as contrast agents for optical imaging of tumors.

Animals↗
PubMed2005

Real time in vivo non-invasive optical imaging using near-infrared fluorescent quantum dots.

RATIONALE AND OBJECTIVE: Deep-tissue optical imaging is of particular interest, as the equipment costs are lower than for competing technologies such as MRI. For this purpose, the development of novel contrast agents with near-infrared (NIR) fluorescence is especially important. We report on the use of NIR semiconductor nanocrystals in deep-tissue in vivo optical imaging. MATERIALS AND METHODS: Semiconductor nanocrystals of CdMnTe/Hg were grown in aqueous solution and then coated with bovine serum albumin (BSA). The nanocrystals were approximately 5 nm in diameter and have a broad fluorescence peak in the NIR (770 nm). Nanocrystals were injected either subcutaneously or intravenously into athymic NCR NU/NU and C3H/HENCR MTV mice and then excited with a spatially broad 633 nm source; the resulting fluorescence was captured with a sensitive CCD camera. RESULTS: We have demonstrated that the nanocrystals are a useful angiographic contrast agent for vessels surrounding and penetrating a murine squamous cell carcinoma in a C3H mouse. Preliminary assessment of the depth of penetration for excitation and emission was done by imaging a beating mouse heart, both through an intact thorax and after a thoracotomy. The temporal resolution associated with imaging the nanocrystals in circulation has been addressed, and the blood clearance for this contrast agent has also been measured. CONCLUSIONS: There was no significant photobleaching or degradation of the nanocrystals after an hour of continuous excitation. The stability of the nanocrystals together with the time resolution of the optical detection makes them particularly attractive candidates for pharmacokinetic imaging studies.

Animals↗
PubMed2005

Optical imaging of functional organization of V1 and V2 in marmoset visual cortex.

Using optical imaging of intrinsic cortical signals, we examined the functional organization of visual cortical areas V1 and V2 of the marmoset (Callithrix jacchus). Previous studies have reported that adult marmosets do not have ocular dominance columns (ODCs); however, recent studies have called this into question. Using optical imaging methods, we examined whether ODCs could be detected in adult marmosets. We found evidence for functional ODCs in some marmosets but not in others. The activation patterns, when present, were relatively weak and appeared as a mosaic of irregular bands or islands. Consistent with studies in other New World monkeys, these data suggest the presence of ODC variability within the marmoset population. Orientation maps in V1 revealed iso-orientation domains organized in semicontinuous bands oriented orthogonal to the V1/V2 border, a pattern unlike that in Macaque monkey. The presence of directional preference maps in V1 was also suggested. In V2, similar to V2 in Macaque monkeys, stripe-like regions of orientation selectivity overlay the pale cytochrome oxidase regions of V2; zones not selective for orientation overlay the cytochrome thin stripes. However, unlike Macaques, we did not observe clear evidence for orientation maps overlying thick cytochrome oxidase stripes. In sum, our data suggest that significant organizational differences exist between the organization of V1 and V2 in the marmoset and that of Old World primates. Implications for the establishment of functional ocular dominance columns, the coestablishment of multiple featural maps, and cortical magnification factors are discussed.

Animals↗
PubMed1999

Near-infrared optical imaging of protease activity for tumor detection.

PURPOSE: To build and test an optical imaging system that is sensitive to near-infrared fluorescent molecular probes activated by specific enzymes in tumor tissues in mice. MATERIALS AND METHODS: The imaging system consisted of a source that delivered 610-650-nm excitation light within a lighttight chamber, a 700-nm longpass filter for selecting near-infrared fluorescence emission photons from tissues, and a charge-coupled device (CCD) for recording images. The molecular probe was a biocompatible autoquenched near-infrared fluorescent compound that was activated by tumor-associated proteases for cathepsins B and H. Imaging experiments were performed 0-72 hours after intravenous injection of the probe in nude mice that bore human breast carcinoma (BT-20). RESULTS: The imaging system had a maximal spatial resolution of 60 microns, with a field of view of 14 cm2. The detection threshold of the nonquenched near-infrared fluorescent dye was subpicomolar in the imaging phantom experiments. In tissue, 250 pmol of fluorochrome was easily detected during the 10-second image acquisition. After intravenous injection of the probe into the tumor-bearing animals, tumors as small as 1 mm became detectable because of tumor-associated enzymatic activation of the quenched compound. CONCLUSION: Tumor proteases can be used as molecular targets, allowing visualization of millimeter-sized tumors. The development of this technology, probe design, and optical imaging systems hold promise for molecular imaging, cancer detection, and evaluation of treatment.

Adenocarcinoma↗
PubMed2003

Shedding light on brain mapping: advances in human optical imaging.

Several functional brain imaging techniques have been used to study human cortical organization. Optical imaging of intrinsic signals (OIS) offers perhaps the best combination of spatial coverage, resolution and speed for mapping the functional topography of human cortex. In this review, we discuss recent advances in optical imaging technology and methodology that have made human OIS easier to implement and more accessible, including improvements in detector characteristics and the development of sophisticated algorithms for reducing motion artifact. Moreover, we discuss how these advances have helped enhance our understanding of the functional organization of the human brain. We also review newly developed analyses for interpreting and validating optical signals, including refined signal analysis techniques and multimodality comparisons. Combined, these advances have enabled the study of not only primary sensory and motor cortices, but also higher cognitive processes such as language production and comprehension. Continued improvement and implementation of this technique promises to shed new light on the functional organization of human cortex.

Algorithms↗
PubMed2006

Optical imaging of experimental arthritis using allogeneic leukocytes labeled with a near-infrared fluorescent probe.

PURPOSE: The purpose of this study was to assess the feasibility of inflammation detection in an antigen-induced arthritis model using fluorescent leukocytes and optical imaging. METHODS: Antigen-mediated monoarthritis was induced in the right knee of 12 Sprague-Dawley rats. Six rats remained untreated and six rats were treated with cortisone. All rats received ex vivo fluorescent-labeled rat leukocytes. Optical images of both knees were acquired before and at 5 min, 1 h, 4 h, and 24 h after cell injection. Images were evaluated qualitatively and quantitatively by calculating signal intensity ratios between the right arthritic (A) and contralateral normal (N) knee. A/N ratios were tested for significant differences between baseline values and values after cell injection using a paired t test as well as between the untreated and cortisone-treated group using an unpaired t test. Synovial specimens were processed and evaluated for labeled cells with fluorescence microscopy. RESULTS: At 4 h and 24 h p.i., the A/N ratios of untreated arthritic knees showed a significant signal increase compared with baseline values (p<0.05) and a significant difference compared with A/N ratios of cortisone-treated animals (p<0.05). Fluorescent microscopy confirmed the presence of labeled cells in the arthritic synovium. CONCLUSION: Inflammation in antigen-induced arthritis can be detected with ex vivo labeled allogenic leukocytes and optical imaging.

Animals↗
PubMed1999

Effects of osmotic stress on dextran diffusion in rat neocortex studied with integrative optical imaging.

Effects of osmotic stress on dextran diffusion in rat neocortex studied with integrative optical imaging. This study investigated how dextran (Mr = 3,000) diffused in rat cortical slices when the osmolarity of the bathing artificial cerebrospinal fluid was altered by varying the NaCl content. The apparent diffusion coefficient, D*, was measured in the neocortex region using fluorescent molecules and the integrative optical imaging (IOI) method. The main results were: 1) the value of D* in rat neocortex in the isotonic (300 mOsm) artificial cerebrospinal fluid at 34 degrees C was D* = 0.68 +/- 0. 01 x 10(-6) cm2 s-1 (mean +/- SE, n = 78) and it could be changed within minutes by varying the extracellular osmolarity. 2) Hypotonic stresses up to -100 mOsm decreased D* by 35% and were fully reversible when the slices were returned to the isotonic medium. Further hypotonic stress to -150 mOsm caused further decrease in D* but after removal of the stress, D* overshot its control value. 3) Hypertonic stress of +50 mOsm increased D*, but the maximum reversible increase in D* was only 15%. Further hypertonic stress (to +200 mOsm) did not cause any further increase in D* and, after removal of the stress, D* undershot the control value. The changes in D* are thought to be related to volume changes of cells in tissue: hypotonic solutions caused cell swelling, resulting in reduced extracellular space and compressed extracellular matrix so that the dextran diffusion was more hindered. Hypertonic solutions had the opposite effect. Recordings of extracellular field potentials in the hippocampal CA1 region demonstrated that, on return to the isotonic solution after exposure to an extreme hypotonic or hypertonic stress, the neurons retained their ability to generate synaptic responses.

Animals↗
PubMed2006

Optical image processing using the photoinduced anisotropy of pyrrylfulgide.

A synthesized photochromic compound-pyrrylfulgide--is prepared as a thin film doped in a polymethylmethacrylate (PMMA) matrix. Under irradiation by UV light, the film converts from the bleached state into a colored state that has a maximum absorption at 635 nm and is thermally stable at room temperature. When the colored state is irradiated by a linearly polarized 650 nm laser, the film returns to the bleached state; photoinduced anisotropy is produced during this process. Application of optical image processing methods using the photoinduced anisotropy of the pyrrylfulgide/PMMA film is described. Examples in non-Fourier optical image processing, such as contrast reversal and image subtraction and summation, as well as in Fourier optical image processing, such as low-pass filtering and edge enhancement, are presented.

Anisotropy↗
PubMed1998

Near-field optical imaging of unstained bacteria: comparison with normal atomic force and far-field optical microscopy in air and aqueous media.

Simultaneous near-field scanning optical and atomic force imaging of bacteria is presented. The bacteria imaged in these studies were unstained. The near-field optical images had excellent signal-to-noise and showed excellent contrast even in these unstained specimens. The images obtained were interpreted in terms of the images that have been obtained by transmission electron microscopy and X-ray imaging. The results show that bacterial near-field optical imaging is going to be a very important tool in the arsenal of the bacteriologist both in terms of understanding the fundamental processes in the life cycle of bacteria with and without cytochemical staining and in terms of clinical diagnostic applications.

Bacillus megaterium↗
PubMedDate not supplied

Detection of dual-gene expression in arteries using an optical imaging method.

We evaluate the in vivo use of an optical imaging method to detect the vascular expression of green fluorescent protein (GFP) or red fluorescent protein (RFP), and to detect the simultaneous expression of GFP and RFP after transduction into arteries by a dual-promoter lentiviral vector driving their concurrent expression. This method involves using a charge-coupled device camera to detect fluorescence, a fiber optic probe to transmit light, and optical filters to distinguish each marker. In animal models, these vectors are locally delivered to target arteries, whereas the gene for a nonfluorescent cell-surface protein is transduced into contralateral arteries as the sham control. The images show distinct areas of bright fluorescence from GFP and RFP along the target arteries on excitation; no exogenous fluorescence is observed in the controls. Measured signal intensities from arteries transduced with the single- and dual-promoter vectors exceed the autofluorescence signal from the controls. Transgene expression of GFP and RFP in vivo is confirmed with confocal microscopy. We demonstrate the use of an optical imaging method to concurrently detect two distinct fluorescent proteins, potentially permitting the expression of multiple transgenes and their localization in the vasculature to be monitored.

Animals↗
PubMed1997

Localized near-infrared spectroscopy and functional optical imaging of brain activity.

Changes in cerebral blood flow (CBF) and cerebral metabolic rates (CMRO2) have been used as indices for changes in neuronal activity. Near-infrared spectroscopy (NIRS) can also measure cerebral haemodynamics and metabolic changes, enabling the possible use of multichannel recording of NIRS for functional optical imaging of human brain activity. Spatio-temporal variations of brain regions were demonstrated during various mental tasks. Non-synchronous behaviour of cerebral haemodynamics during the neuronal activation was observed. Gender- and handedness-dependent lateralization of the function between right and left hemispheres was demonstrated by simultaneous measurement using two NIR instruments during the mirror-drawing task. A lack of interhemispheric integration was observed with schizophrenic patients. These observations suggest an application for NIRS in psychiatric disease management, as an addition to clinical monitoring at the bedside. A time resolved 64-channel optical imaging system was constructed. This consisted of three picosecond laser diodes and 64 channels of TAC and CFD systems. Image reconstruction for phantom model systems was performed. Time-resolved quantitative optical imaging will become real in the very near future.

Adult↗
PubMed2004

Live optical imaging of nervous system development.

Although development of the nervous system is inherently a process of dynamic change, until recently it has generally been investigated by inference from static images. However, advances in live optical imaging are now allowing direct observation of growth, synapse formation, and even incipient function in the developing nervous system, at length scales from molecules to cortical regions, and over timescales from milliseconds to months. In this review, we provide technical background and present examples of how these new methods, including confocal and two-photon microscopy, GFP-based markers, and functional indicators, are being applied to provide fresh insight into long-standing questions of neural development.

Animals↗
PubMed2004

Mapping cone- and rod-induced retinal responsiveness in macaque retina by optical imaging.

PURPOSE: To map the distribution of cone- or rod-induced retinal responsiveness by optical imaging from macaque retina. METHODS: The light reflectance changes in the posterior retina after a flash stimulus in anesthetized rhesus monkeys were measured by a modified fundus camera system equipped with a charge-coupled device (CCD) camera. The response topography of the optical signals was obtained in either light- or dark-adapted conditions. RESULTS: With infrared observation light, the whole posterior pole became darkened after the stimulus. The response topography in light-adapted conditions demonstrated a steep peak of darkening at the fovea, together with the gradual decrease of signal intensity away from the fovea toward the periphery. In dark-adapted conditions, the optical signal showed additional peaks along the circular region surrounding the macula at the eccentricity of the optic disc, together with the central peak at the fovea. A statistically significant positive correlation was obtained between the light reflectance changes in infrared observation light and the focal responses in multifocal electroretinogram (mfERG) at the corresponding retinal locations. CONCLUSIONS: The response topography in the retina, obtained by optical imaging, was consistent with psychophysical cone or rod sensitivity in humans and anatomic cone or rod distribution in humans and macaques. The cone- or rod-induced retinal responsiveness within the posterior pole region was noninvasively recorded within a short recording time.

Animals↗
PubMed1992

Optical imaging of architecture and function in the living brain sheds new light on cortical mechanisms underlying visual perception.

Long standing questions related to brain mechanisms underlying perception can finally be resolved by direct visualization of the architecture and function of mammalian cortex. This advance has been accomplished with the aid of two optical imaging techniques with which one can literally see how the brain functions. The upbringing of this technology required a multi-disciplinary approach integrating brain research with organic chemistry, spectroscopy, biophysics, computer sciences, optics and image processing. Beyond the technological ramifications, recent research shed new light on cortical mechanisms underlying sensory perception. Clinical applications of this technology for precise mapping of the cortical surface of patients during neurosurgery have begun. Below is a brief summary of our own research and a description of the technical specifications of the two optical imaging techniques. Like every technique, optical imaging also suffers from severe limitations. Here we mostly emphasize some of its advantages relative to all alternative imaging techniques currently in use. The limitations are critically discussed in our recent reviews. For a series of other reviews, see Cohen (1989).

Brain↗