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Electro-optical imaging of F-actin and endoplasmic reticulum in living and fixed plant cells.

Confocal and video micrographs of living and fixed alfalfa roots, onion epithelial and pear pollen cells illustrate the architecture of the cytoskeleton and endoplasmic reticulum in plant cells. Fixation of plant tissues to preserve cytoplasmic structure poses special problems. When possible, emphasis should be placed on the imaging of structures in stained living cells over time. The early events that occur when Nod factors or bacteria elicit nodule formation in alfalfa roots will illustrate several approaches to plant cell fixation, staining and imaging. The first observable events after Nod factor stimulation occur in root hairs and are changes in rates of cytoplasmic streaming, nuclear movements, and changes in the shape of the vacuole. Within ten minutes, the endoplasmic reticulum shifts position towards the tip of the root hair. For comparison, the endoplasmic reticulum localization in pollen tubes and onion epithelial cells will be illustrated. The actin cytoskeleton undergoes a series of changes over a twelve hour period. These changes in the cytoskeleton are spatially and temporally correlated with the observed growth changes of the root hairs. This dynamic change of the actin filament and endoplasmic reticulum and associated secretory vesicles in these root hairs suggests a mechanism for the observed root hair growth changes.

Actins↗

Time-resolved imaging of optical coefficients through murine chest cavities.

As small animal optical imaging and tomography are gaining popularity for interrogating functional and molecular events in vivo, it becomes increasingly necessary to gain knowledge of the optical properties of the species investigated to better understand and describe photon propagation through their tissues. To achieve characterization of the spatial variation of average optical properties through murine chest cavities, time- and spatially resolved measurements of femto-second laser pulse transmission are performed through mice using a high-speed gated image intensifier. Application of time-resolved diffusion theory for finite slab geometry is first confirmed on phantoms and then applied to in vivo measurements for spatially resolving and quantifying mouse optical properties. Photon transmission images through mouse chest cavities are further obtained at different time gates to visualize the spatial variation observed and confirm the optical coefficient patterns calculated.

Animals↗

Computational methods for analysis of human breast tumor tissue in optical coherence tomography images.

Optical coherence tomography (OCT) has been demonstrated as a promising means of identifying the boundaries between normal and diseased breast tissue. This capability has yielded promise for the development of OCT techniques for biopsy guidance, surgical margin assessment, and minimally invasive evaluation of disease states. We present methods for the assessment of human breast tissue based on spatial and Fourier-domain analysis. Derived from preliminary OCT data, these methods are aimed at the development of automated diagnostic tools that will aid in the translation of this technology into the clinical environment.

Algorithms↗

Bedside imaging of intracranial hemorrhage in the neonate using light: comparison with ultrasound, computed tomography, and magnetic resonance imaging.

Medical optical imaging (MOI) uses light emitted into opaque tissues to determine the interior structure. Previous reports detailed a portable time-of-flight and absorbance system emitting pulses of near infrared light into tissues and measuring the emerging light. Using this system, optical images of phantoms, whole rats, and pathologic neonatal brain specimens have been tomographically reconstructed. We have now modified the existing instrumentation into a clinically relevant headband-based system to be used for optical imaging of structure in the neonatal brain at the bedside. Eight medical optical imaging studies in the neonatal intensive care unit were performed in a blinded clinical comparison of optical images with ultrasound, computed tomography, and magnetic resonance imaging. Optical images were interpreted as correct in six of eight cases, with one error attributed to the age of the clot, and one small clot not seen. In addition, one disagreement with ultrasound, not reported as an error, was found to be the result of a mislabeled ultrasound report rather than because of an inaccurate optical scan. Optical scan correlated well with computed tomography and magnetic resonance imaging findings in one patient. We conclude that light-based imaging using a portable time-of-flight system is feasible and represents an important new noninvasive diagnostic technique, with potential for continuous monitoring of critically ill neonates at risk for intraventricular hemorrhage or stroke. Further studies are now underway to further investigate the functional imaging capabilities of this new diagnostic tool.

Animals↗

In vivo detection of small subsurface melanomas in athymic mice using noninvasive fiber optic confocal imaging.

Fiber optic confocal imaging, following intravenous administration of fluorescently labeled antibodies and Texas Red-dextran, enabled in vivo detection of melanoma and surrounding blood vessels in athymic mice. Human melanoma cells (three cell lines) and cultured normal human skin cells were implanted intradermally into the haunch skin of anesthetized athymic BALB/C mice and allowed to grow to a maximum size of 2 mm diameter. Using three different fluorescein-isothiocyanate-labeled antimelanoma antibodies, single channel confocal images of melanoma cells were obtained in vivo. Using noninvasive techniques, the overall in vivo melanoma detection rate for tumors within 0.2 mm of the skin surface was 84% (27 of 32 tumors). Normal cultured human skin cells were found to have little or no fluorescence after administration of the fluorescein-isothiocyanate-labeled antibodies and tumors were not labeled by an isotype control antibody. Dual channel imaging of the implanted melanoma tumor and surrounding dermal vasculature in vivo showed increased blood vessel density at the melanoma site. Conventional immunoperoxidase histology confirmed that fiber optic confocal imaging was able to detect melanoma tumors up to 0.2 mm below the skin surface, in vivo.

Animals↗

Optical biopsy and imaging using optical coherence tomography.

Optical coherence tomography is a new imaging technique that can perform high-resolution, micrometre-scale, cross-sectional imaging in biological systems. The technology has been developed, and reduced to, preliminary clinical practice in ophthalmology. The challenging problem that OCT may address is the development of 'optical biopsy' techniques. These techniques can provide diagnostic imaging of tissue morphology without the need for excision of specimens. Many investigations remain to identify optimal areas for clinical application, and additional engineering must be done to integrate vertically the technology and to reduce it to clinical practice. Nevertheless, preliminary studies indicate the feasibility of developing this technology for a wide range of clinical and research diagnostic imaging applications. The ability to non-excisionally evaluate tissue morphology using a catheter or an endoscope could have a significant impact on the diagnosis and management of a wide range of diseases.

Arteries↗

Overview of functional imaging.

Provides an overview of the many different modalities of imaging techniques, including positron emission tomography, single-photon emission computed tomography, functional magnetic resonance imaging, optical imaging, and magnetic source imaging.

Brain Neoplasms↗

Optical coherence tomography for ultrahigh resolution in vivo imaging.

Optical coherence tomography (OCT) is an emerging biomedical optical imaging technique that performs high-resolution, cross-sectional tomographic imaging of microstructure in biological systems. OCT can achieve image resolutions of 1-15 microm, one to two orders of magnitude finer than standard ultrasound. The image penetration depth of OCT is determined by the optical scattering and is up to 2-3 mm in tissue. OCT functions as a type of 'optical biopsy' to provide cross-sectional images of tissue structure on the micron scale. It is a promising imaging technology because it can provide images of tissue in situ and in real time, without the need for excision and processing of specimens.

Arteries↗

Molecular imaging: state of the art.

Molecular imaging includes all imaging methods applied in the identification, characterization and assessment "in vivo" of biological processes which occur at the cellular and molecular level. Molecular imaging parallels the remarkable advances achieved in the medical field, culminated in the sequencing of human DNA, the genome project. The understanding of the genetic basis of diseases and of human biology in general, together with the development of new drugs, led to a growing need for novel, sensitive and safe imaging technology to be rapidly translated from animal models into patients. The present evaluation of presence/absence of disease is based on anatomic and morphologic changes, the result of underlying molecular alterations. Direct "in vivo" visualization of these alterations allows early diagnosis before the onset of typical pathologic manifestations. Similarly, short-term effects of therapy can be directly visualized. In a near future, "evidence-based" medicine will become "presymptomatic" medicine. Histopathology will be replaced by "molecular pathology" with genomic implications in disease classification. In the near future the conventional morphologic methods should be supported and then replaced by the new functional and molecular methods with additional information proved useful for the diagnostic approach. In this article, the potentialities and applications of Nuclear Medicine, Magnetic Resonance Imaging, Optical Imaging and Contrast-enhanced Ultrasound are reviewed.

Diagnostic Imaging↗

Molecular imaging perspectives.

Molecular imaging is an emerging technology at the life science/physical science interface which is set to revolutionize our understanding and treatment of disease. The tools of molecular imaging are the imaging modalities and their corresponding contrast agents. These facilitate interaction with a biological target at a molecular level in a number of ways. The diverse nature of molecular imaging requires knowledge from both the life and physical sciences for its successful development and implementation. The aim of this review is to introduce the subject of molecular imaging from both life science and physical science perspectives. However, we will restrict our coverage to the prominent in vivo molecular imaging modalities of magnetic resonance imaging, optical imaging and nuclear imaging. The physical basis of these imaging modalities, the use of contrast agents and the imaging parameters of sensitivity, temporal resolution and spatial resolution are described. Then, the specificity of contrast agents for targeting and sensing molecular events, and some applications of molecular imaging in biology and medicine are given. Finally, the diverse nature of molecular imaging and its reliance on interdisciplinary collaboration is discussed.

Contrast Media↗

Tomorrow's radiologist: what future?

Today's radiology is experiencing two major trends, one negative and one positive. The first is the so-called turf war, in other words, the progressive invasion of the imaging domain by other specialists such as cardiologists, urologists, gastroenterologists, gynaecologists etc. who are taking over various techniques from ultrasonography (US) to computed tomography (CT) to magnetic resonance imaging (MRI). In this process, they are aided by new technologies such as picture archiving and communication systems (PACS) and computed-aided diagnosis CAD and by radiology technologists who collaborate with them, replacing radiologists. The positive aspect is the outstanding technological evolution: the advent of molecular imaging, optical imaging, nanotechnologies, teleradiology and percutaneous gene therapy. While dramatically expanding the diagnostic possibilities down to the subcellular level, these techniques demand new forms of training in radiology and interdisciplinary cooperation. Tomorrow's radiologist will need to acquire appropriate clinical knowledge, restore contact with the patient to take on a prominent role in the diagnostic process, learn the basic sciences, foster a multidisciplinary approach and finally be able to use the Internet for learning and continuing education. Tomorrow's radiologists will survive if they learn to reinvent themselves.

Biomedical Research↗

Optimizing imaging polarimeters constructed with imperfect optics.

Imaging polarimeters are often designed and optimized by assuming that the polarization properties of the optics are nearly ideal. For example, we often assume that the linear polarizers have infinite extinction ratios. It is also usually assumed that the retarding elements have retardances that do not vary either spatially or with the angle of incidence. We consider the case where the polarization optics used to develop an imaging polarimeter are imperfect. Specifically, we examine the expected performance of a system as the extinction ratio of the diattenuators degrades, as the retardance varies spatially, and as the retardance varies with incidence angle. It is found that the penalty in the signal-to-noise ratio for using diattenuators with low extinction ratios is not severe, as an extinction ratio of 5 causes only a 2.0 dB increase in the noise in the reconstructed Stokes parameter images compared with an ideal diattenuator. Likewise, we find that a system can be optimized in the presence of spatially varying retardance, but that angular positioning error is far more important in rotating retarder imaging polarimeters.

Journal Article↗

Optic nerve diffusion tensor imaging in optic neuritis.

Diffusion tensor magnetic resonance imaging (DT-MRI) provides in vivo information about the pathology of multiple sclerosis lesions. Increases in mean diffusivity (MD) and reductions in fractional anisotropy (FA) have been found and may represent axonal disruption. The optic nerve is an ideal structure for study by DT-MRI but previous clinical studies did not obtain the full diffusion tensor necessary to calculate MD and FA. In this study, a technique that was specifically developed to achieve full diffusion tensor measurements from the optic nerve (zonal oblique multislice (ZOOM) echoplanar imaging) was applied to 25 patients with a single unilateral episode of optic neuritis at least one year previously, and 15 controls. The intraorbital nerves were segmented on non-diffusion-weighted images and the regions of interest transferred to MD, FA, and eigenvalue maps to obtain quantitative data. Quantitative visual testing and electrophysiology were also performed. In affected nerves, mean MD and mean orthogonal eigenvalue lambda( perpendicular) were elevated, and mean FA reduced compared with clinically unaffected contralateral nerves (P < 0.001) and control nerves (P < 0.001). The mean principal eigenvalue lambda\\ was significantly increased in affected nerves compared to contralateral unaffected nerves (P = 0.04) but not compared to control nerves (P = 0.13). There was no association of clinical measures of visual function in affected eyes with the DT-MRI parameters but there was a significant correlation of the whole field visual evoked potential (VEP) amplitude with MD (r = -0.57, P = 0.006) and lambda( perpendicular) (r = -0.56, P = 0.007). These findings suggest that optic nerve DT-MRI measures provide an indication of the structural integrity of axons.

Adult↗

Review of tissue simulating phantoms for optical spectroscopy, imaging and dosimetry.

Optical spectroscopy, imaging, and therapy tissue phantoms must have the scattering and absorption properties that are characteristic of human tissues, and over the past few decades, many useful models have been created. In this work, an overview of their composition and properties is outlined, by separating matrix, scattering, and absorbing materials, and discussing the benefits and weaknesses in each category. Matrix materials typically are water, gelatin, agar, polyester or epoxy and polyurethane resin, room-temperature vulcanizing (RTV) silicone, or polyvinyl alcohol gels. The water and hydrogel materials provide a soft medium that is biologically and biochemically compatible with addition of organic molecules, and are optimal for scientific laboratory studies. Polyester, polyurethane, and silicone phantoms are essentially permanent matrix compositions that are suitable for routine calibration and testing of established systems. The most common three choices for scatters have been: (1.) lipid based emulsions, (2.) titanium or aluminum oxide powders, and (3.) polymer microspheres. The choice of absorbers varies widely from hemoglobin and cells for biological simulation, to molecular dyes and ink as less biological but more stable absorbers. This review is an attempt to indicate which sets of phantoms are optimal for specific applications, and provide links to studies that characterize main phantom material properties and recipes.

Biomimetics↗