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Near-infrared fluorescence optical imaging and tomography.

The advent of recent advances in near-infrared laser diodes and fast electro-optic detection has spawned a new research field of diagnostic spectroscopy and imaging based on targeting and reporting exogenous fluorescent agents. This review seeks to concisely address the physics, instrumentation, advancements in tomography, and near-infrared fluorescent contrast agent development that promises selective and specific molecular targeting of diseased tissues. As an example of one area of the field, recent work focusing on pharmacokinetic analysis of fluorophores targeting the epidermal growth factor receptor (EGFR) is presented in a human breast cancer xenograft mouse model to demonstrate specificity of molecularly targeted contrast agents. Finally, a critical evaluation of the limitations and the opportunities for future translation of fluorescence-enhanced optical imaging of deep tissues is presented.

Animals↗

Optical imaging of intrinsic signals in somatosensory cortex.

The methods of optical intrinsic signal (OIS) imaging and microelectrode mapping of single neuron receptive fields (RFs) were used in combination (in the same squirrel monkey or cat) to characterize the spatial and temporal attributes of the response of contralateral SI cortex to cutaneous flutter stimulation. A change in the location of the stimulated skin site was accompanied by a shift in the locus of the SI optical response. The spatial ordering of the optical responses to independent stimulation of each site in an array of skin sites was consistent with the features of SI topographical organization described in published RF mapping studies. While the single neuron RF mapping observations and the optical response obtained at a given time after onset of flutter stimulation always were positively correlated, the degree of correlation improved progressively with time after stimulus onset (the longest stimulus duration used was 10 s). Analysis of the temporal development of the optical response to cutaneous flutter stimulation revealed that not only does absorbance increase to attain a maximum in the SI region which receives its main input from the stimulated skin site, but at the same time absorbance declines to below-background values in an extensive region of surrounding cortex. The results are interpreted to indicate that the pattern of SI activity evoked by a cutaneous flutter stimulus exhibits increasing spatial contrast (becomes progressively more distinguishable from the activity of surrounding cortex) over periods of continuous stimulation at least as long as 10 s. This time-dependent 'funneling' of the SI spatial activity pattern is proposed to underlie the prominent enhancement of human spatial discriminative capacity which occurs (e.g. Physiol. Behav. 5 (1970) 1431) when oscillatory tactile stimuli are used.

Animals↗

Specifying and controlling the optical image on the human retina.

A review covering the trends that led to the current state of knowledge in the areas of: (a) schematic models of the eye, and the definition of the retinal image in terms of first-order optics; (b) the description of the actual image on the retina and methods for accessing and characterizing it; (c) available procedures for controlling the quality of the retinal image in defined situations; and (d) intra-receptoral optical effects that cause differences between the light distribution on the retinal surface and at the level of interaction with photopigment molecules.

Humans↗

Noninvasive optical imaging of the subarachnoid space and cerebrospinal fluid pathways based on near-infrared fluorescence.

The authors have developed a noninvasive optical method to image the subarachnoid space and cerebrospinal fluid pathways in vivo based on the near-infrared fluorescence of indocyanine green (ICG). The ICG was bound to purified lipoproteins (ICG-lipoprotein) and injected into the subarachnoid space of neonatal and adult rats. The ICG fluorescence was detected by a cooled charge-coupled device camera. After injection of ICG-lipoprotein into the cerebral subarachnoid space of the neonatal rat, ICG fluorescence was clearly detected at the injection site through the skull and skin. The ICG fluorescence was observed in the cerebellum and the lumbar spinal cord 1 and 8 hours postinjection, respectively. After injection of ICG-lipoprotein into the lumbar spinal subarachnoid space of an adult rat, ICG fluorescence was observed from the injection site to the thoracic levels along the spinal subarachnoid space. In addition, with the rat's head tilted downward, ICG fluorescence had extended to the cerebral subarachnoid space by 1 hour postinjection. The ICG fluorescence imaging of the cerebral subarachnoid space demonstrated an increase in fluorescence intensity around the lambdoid suture and the forebrain. On dissection of the rat brain the former location was identified as the supracerebellar cistern and the latter as the olfactory cistern. The results of this study are the first to demonstrate that an optical technique is applicable to imaging of the subarachnoid space and cerebrospinal fluid pathways in vivo. In addition, ICG-lipoprotein provides a sensitive optical tracer for imaging extravascular biological structures. Finally, ICG fluorescence imaging does not require an intricate imaging system because ICG is localized near the surface of the body.

Animals↗

Quality parameters analysis of optical imaging systems with enhanced focal depth using the Wigner distribution function

An analysis of the Strehl ratio and the optical transfer function as imaging quality parameters of optical elements with enhanced focal length is carried out by employing the Wigner distribution function. To this end, we use four different pupil functions: a full circular aperture, a hyper-Gaussian aperture, a quartic phase plate, and a logarithmic phase mask. A comparison is performed between the quality parameters and test images formed by these pupil functions at different defocus distances.

Journal Article↗

Attention and probability effects in the human occipital cortex: an optical imaging study.

A new imaging technique (event-related optical signal, EROS) reveals the time course of neural activity in selected cortical areas of normal human subjects. This technique was used to study the event-related activity in striate and extrastriate occipital areas in an experiment in which spatial selective attention and stimulus probability were manipulated. The results show that attention effects are evident in the initial response in extrastriate cortex (latency < 100 ms), but not in striate cortex, confirming previous modeling effects. They also show that the initial response in striate cortex is modulated by stimulus probability, suggesting the occurrence of pre-attentive memory phenomena in primary visual cortex.

Adult↗

Looking and listening to light: the evolution of whole-body photonic imaging.

Optical imaging of live animals has grown into an important tool in biomedical research as advances in photonic technology and reporter strategies have led to widespread exploration of biological processes in vivo. Although much attention has been paid to microscopy, macroscopic imaging has allowed small-animal imaging with larger fields of view (from several millimeters to several centimeters depending on implementation). Photographic methods have been the mainstay for fluorescence and bioluminescence macroscopy in whole animals, but emphasis is shifting to photonic methods that use tomographic principles to noninvasively image optical contrast at depths of several millimeters to centimeters with high sensitivity and sub-millimeter to millimeter resolution. Recent theoretical and instrumentation advances allow the use of large data sets and multiple projections and offer practical systems for quantitative, three-dimensional whole-body images. For photonic imaging to fully realize its potential, however, further progress will be needed in refining optical inversion methods and data acquisition techniques.

Image Enhancement↗

Optical imaging of epileptiform activity in human neocortex.

The surgical outcomes of patients suffering from neocortical epilepsy are not as successful as the surgical outcomes from resections of epilepsy patients with mesial temporal sclerosis. The main difficulty in the treatment of neocortical epilepsy is that current technology has limited accuracy in mapping neocortical epileptogenic tissue. It is known that the optical spectroscopic properties of brain tissue are correlated with changes in neuronal activity. The method of mapping these activity-evoked optical changes is known as imaging of intrinsic optical signals (IIOS). Activity-evoked optical changes measured in neocortex are generated by changes in cerebral hemodynamics (i.e., changes in blood oxygenation and blood volume). Our experimental approach was to acquire high-resolution IIOS maps of epileptiform activity in patients undergoing surgery for medically intractable neocortical epilepsy. Both spontaneous and stimulation-evoked epileptiform activity was monitored. Imaging of intrinsic optical signals was able to localize neocortical epileptic foci precisely by using changes in blood volume in contrast to changes in blood oxygenation. IIOS has the potential to translate from a purely research tool to a new intraoperative approach for the surgical treatment of neocortical epilepsy.

Blood Volume↗

Mirror-image optic nerve dysplasia with associated anisometropia in identical twins.

BACKGROUND: Monozygotic (MZ) or "identical" twins arise from a single fertilized egg, which divides into two embryos at an early stage of development. As a result, MZ twins have identical genomes and are always of the same sex. METHODS: A case of optic nerve hypoplasia and anisometropia, in association with mirror-image presentation in a set of 12-year-old identical twins, is reported. The monozygotic twinning event responsible for identical twins--as well as the rare phenomenon of mirror imaging--is described. RESULTS: The combined occurrence of anisometropia and optic nerve hypoplasia in mirror-image presentation in a set of monozygotic twins provides a unique opportunity to study the genetic versus environmental influences on the development of optic nerve hypoplasia. CONCLUSIONS: Although the cause of optic nerve hypoplasia remains unclear, its associated mirror-image presentation in this case suggests a possible genetic predisposition.

Anisometropia↗

High-resolution optical imaging from trajectory time distributions.

Trajectory time distribution optical microscopy (TTDOM), which records the mean off-times of single molecular fluorescent indicators that light up when they collide with vesicles, is extended to record fluorescence durations or on-times. TTDOM can distinguish shapes of objects that are smaller than the diffraction limited resolution. The fluorescence duration time image can also provide high-resolution information. The effects of the threshold that separates fluorescent bursts from background signals and of two or more probes visiting the vesicles simultaneously have been investigated systematically. New experimental results along with simulations indicate that TTDOM is capable of providing the size and shape of objects and information on probe-vesicle binding.

Dimyristoylphosphatidylcholine↗

High-resolution nonlinear optical imaging of live cells by second harmonic generation.

By adapting a laser scanning microscope with a titanium sapphire femtosecond pulsed laser and transmission optics, we are able to produce live cell images based on the nonlinear optical phenomenon of second harmonic generation (SHG). Second harmonic imaging (SHIM) is an ideal method for probing membranes of living cells because it offers the high resolution of nonlinear optical microscopy with the potential for near-total avoidance of photobleaching and phototoxicity. The technique has been implemented on three cell lines labeled with membrane-staining dyes that have large nonlinear optical coefficients. The images can be obtained within physiologically relevant time scales. Both achiral and chiral dyes were used to compare image formation for the case of single- and double-leaflet staining, and it was found that chirality plays a significant role in the mechanism of contrast generation. It is also shown that SHIM is highly sensitive to membrane potential, with a depolarization of 25 mV resulting in an approximately twofold loss of signal intensity.

3T3 Cells↗

[Assessment of the optical image quality of the eye using raytracing technique of corneal topography data].

BACKGROUND: Optical aberrations in the optical system may downgrade image quality and cannot be fully compensated by spherocylindrical glasses. The subjectively evaluated visual acuity may be significantly reduced. The purpose of this study was to calculate the image forming properties of the eye using a spotlight source or alternatively extended objects. METHODS: A convex and first derivative continuous (C1) surface from the rough height data of the anterior corneal surface (TMS-1, Tomey, Erlangen) or the anterior and posterior corneal surface (Orbscan, Orbtec, USA) was calculated by means of an interpolating subdivision scheme (modified Butterfly algorithm). The characteristics of the residual refractive surfaces were used according to Navarro's eye model. The focal distance was calculated from the exact raytracing calculation (Snellius' law) of the point-spread function by minimising the variance of the point-spread function. The diffraction property of the aperture stop was implemented with a transmission characteristic according to a radially symmetrical Bessel function within the entrance pupil. The algorithm was realised with a C code on the LINUX platform and applied to a normal eye (example 1, TMS-1), an eye with severe keratoconus (example 2, TMS-1) and an eye with corneal scars (example 3, Orbscan). RESULTS: The focal distance in example 1 (22.5 mm, 22.6 mm, and 22.8 mm) increased with the pupil diameter (2 mm, 3 mm, and 5 mm). The variance of the approximately radially symmetrical point-spread function in the focal plane attained a minimum value with a pupil size of 3 mm (0.164, 0.104, and 0.230). In example 2, the focal distance changed inconclusively (21.1 mm, 21.0 mm, and 21.3 mm) with the pupil size (2 mm, 3 mm, and 5 mm). The variance of the markedly asymmetrical point-spread function in the focal plane was systematically higher compared to the values of example 1 and reached a minimum value with a pupil size of 3 mm (0.255, 0.224, and 0.371). The imaging of the sinus-modulated pattern is anisotropic due to the asymmetry of the point-spread function. In example 3, the focal distance (22.3 mm, 22.3 mm, and 22.5 mm) did not change systematically with the pupil size (2 mm, 3 mm, and 5 mm). The variance of the nearly radially symmetrical point-spread function changed only marginally between pupil sizes of 2 mm and 3 mm (0.231, 0.239, and 0.338). CONCLUSIONS: Raytracing of corneal topography height data based on refined eye models with the option of auto-focussing has the potential to trace the optical resolution of the eye for arbitrary objects. Further studies on contrast sensitivity and the conversion of the real image to a perceived image by the retina and brain are required for complete modeling of subjective visual acuity.

Adult↗

Detector Concept for OPET-A Combined PET and Optical Imaging System.

The design of an imaging system capable of detecting both high-energy γ-rays and optical wavelength photons is underway at the UCLA Crump Institute for Molecular Imaging. This system, which we call optical PET (OPET), will be capable of non-invasively and repeatedly imaging small animal models in vivo for the presence of PET and optical signals. In this study, we describe the physical principles behind the operation of the OPET imaging system and discuss the design concept for one of the detector modules. Additionally, we demonstrate the operation of an initial prototype detector module for simultaneous detection and imaging of annihilation radiation and single optical photons emanating from separate sources. These results indicate that the construction of an imaging system based on this detector technology is feasible.

Journal Article↗

Optical imaging of odor preference memory in the rat olfactory bulb.

Early olfactory preference learning in rat pups occurs when novel odors are paired with reinforcing tactile stimulation that activate the noradrenergic locus coeruleus. Pairing of odor and a noradrenergic agonist in the olfactory bulb is both necessary and sufficient for odor preference learning. This suggests the memory change occurs in the olfactory bulb. Previous electrophysiological experiments demonstrated that odor preference training induces an increase in the field excitatory postsynaptic potential to olfactory nerve input and an alteration, after training, in glomerular [14C]2- deoxyglucose uptake and in single-unit responses of principal cells. We investigate here whether, 24 h after olfactory preference training, there is an alteration in intrinsic optical signals at the glomerular level. Six-day-old rat pups were trained, as previously, for a peppermint odor preference. Trained pups and control littermates were subjected to imaging of odor-induced intrinsic optical signals 1 day after the training session. Trained pups exhibited significantly larger responses to the peppermint compared with untrained littermates previously exposed to the same odor. The response of trained pups to a control odor (amyl acetate) was, however, not significantly different from that of untrained littermates. These observations demonstrate that odor preference memory can be read-out by optical imaging techniques.

Animals↗

Receptor-targeted optical imaging of tumors with near-infrared fluorescent ligands.

We report here the in vivo diagnostic use of a peptide-dye conjugate consisting of a cyanine dye and the somatostatin analog octreotate as a contrast agent for optical tumor imaging. When used in whole-body in vivo imaging of mouse xenografts, indotricarbocyanine-octreotate accumulated in tumor tissue. Tumor fluorescence rapidly increased and was more than threefold higher than that of normal tissue from 3 to 24 h after application. The targeting conjugate was also specifically internalized by primary human neuroendocrine tumor cells. This imaging approach, combining the specificity of ligand/receptor interaction with near-infrared fluorescence detection, may be applied in various other fields of cancer diagnosis.

Animals↗

Nanoscale optical imaging of chromosomes with apertureless microscopy.

Near-field optical structure of the centromere region of undyed polytene chromosomes has been observed using an apertureless near-field optical microscope that detects the intensity of light scattered from an atomic force microscope tip under laser illumination. The centromere is of primary importance to the functioning of the chromosome in the cell during cell division. It is also particularly interesting for structural/optical studies since its DNA repeat sequences are highly conserved among organisms and it is possible that they play a part in the centromere self assembly (Clark and Wall 1996).

Animals↗