Studies of images of short-lived events using ERTS data
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The search for life in our solar system relies on our ability to measure key biomarkers in situ and identify them as likely of biotic origin. One class of biomarkers that would be indicative of life is cell-like features such as the colocalization of key biochemicals, spatial organization of native UV fluorescent compounds, and polarization across membrane compartments. The Europa Lander Science Definition Team has emphasized the need for a light microscope to search for these features on Europa, and the Europa Light Microscope (ELM) has been developed to address this need. This work presents the development towards high launch readiness of ELM by demonstrating its optical resolution under brightfield illumination, deep UV fluorescence imaging of unstained living samples, and multicolor imaging of samples stained with exogenous fluorophores targeting proteins, lipids, and nucleic acids. This work also seeks to extend the capabilities of ELM by incorporating fluorescence-based assays for membrane potential as a key biomarker for living cellular material.
The methods utilized to perform an infrared thermography inspection of the bondline of structural reinforcing caps on X-33 liquid hydrogen (LH2) tank during the manufacture are described in this presentation. A total of thirty-six caps were bonded to the tank using a wet cobond process with vacuum bagging and low temperature curing. The thermal map of each bondline was acquired through the vacuum bag by applying heat to the cap and then viewing it during cool down. Poorly bonded regions, or voids, were characterized by localized hot spots in the thermal image. These regions were marked on the tank by referring to a live thermal image, then hand massaged in an attempt to remove the entrapped air. The cap was then reinspected and, if it was determined that the entrapped air could not be removed, the cap was removed and the adhesive cleaned away before it cured. Measures were then taken to better match the surface of the cap to the tank and the process repeated. After the bondline had cured sufficiently to allow vacuum bag removal, a "flash heating" thermographic Investigation was performed to verify that the cap had bonded properly to the tank. Any regions that had opened up after the preliminary inspection were marked and filled by drilling small holes in the cap and injecting resin. A final inspection was then performed to ensure the quality of the bondline.
Voyager encounter data are presented in computer animation (CA) and real (R) animation. The highlights include a view of 2 full rotations of Neptune. It shows spacecraft trajectory 'diving' over Neptune and intercepting Triton's orbit, depicting radiation and occulation zones. Also shown are a renegade orbit of Triton and Voyager's encounter with Neptune's Magnetopause. A model of the spacecraft's complex maneuvers during close encounters of Neptune and Triton is presented. A view from Earth of Neptune's occulation experiment is is shown as well as a recreation of Voyager's final pass. There is detail of Voyager's Image Compensation technique which produces Voyager images. Eighteen images were produced on June 22 - 23, 1989, from 57 million miles away. A 68 day sequence which provides a stroboscopic view - colorization approximates what is seen by the human eye. Real time images recorded live from Voyager on 8/24/89 are presented. Photoclinometry produced the topography of Triton. Three images are used to create a sequence of Neptune's rings. The globe of Neptune and 2 views of the south pole are shown as well as Neptune rotating. The rotation of a scooter is frozen in images showing differential motion. There is a view of rotation of the Great Dark Spot about its own axis. Photoclinometry provides a 3-dimensional perspective using a color mosaic of Triton images. The globe is used to indicate the orientation of Neptune's crescent. The east and west plumes on Triton are shown.
High resolution B-mode ultrasound images of the common carotid artery are obtained with an ultrasound transducer using a standardized methodology. Subjects are supine with the head counter-rotated 45 degrees using a head pillow. The jugular vein and carotid artery are located and positioned in a vertical stacked orientation. The transducer is rotated 90 degrees around the centerline of the transverse image of the stacked structure to obtain a longitudinal image while maintaining the vessels in a stacked position. A computerized methodology assists operators to accurately replicate images obtained over several spaced-apart examinations. The methodology utilizes a split-screen display in which the arterial ultrasound image from an earlier examination is displayed on one side of the screen while a real-time live ultrasound image from a current examination is displayed next to the earlier image on the opposite side of the screen. By viewing both images, whether simultaneously or alternately, while manually adjusting the ultrasound transducer, an operator is able to bring into view the real-time image that best matches a selected image from the earlier ultrasound examination. Utilizing this methodology, measurement of vascular dimensions such as carotid arterial IMT and diameter, the coefficient of variation is substantially reduced to values approximating from about 1.0% to about 1.25%. All images contain anatomical landmarks for reproducing probe angulation, including visualization of the carotid bulb, stacking of the jugular vein above the carotid artery, and initial instrumentation settings, used at a baseline measurement are maintained during all follow-up examinations.
A standardized acquisition methodology assists operators to accurately replicate high resolution B-mode ultrasound images obtained over several spaced-apart examinations utilizing a split-screen display in which the arterial ultrasound image from an earlier examination is displayed on one side of the screen while a real-time "live" ultrasound image from a current examination is displayed next to the earlier image on the opposite side of the screen. By viewing both images, whether simultaneously or alternately, while manually adjusting the ultrasound transducer, an operator is able to bring into view the real-time image that best matches a selected image from the earlier ultrasound examination. Utilizing this methodology, dynamic material properties of arterial structures, such as IMT and diameter, are measured in a standard region over successive image frames. Each frame of the sequence has its echo edge boundaries automatically determined by using the immediately prior frame's true echo edge coordinates as initial boundary conditions. Computerized echo edge recognition and tracking over multiple successive image frames enhances measurement of arterial diameter and IMT and allows for improved vascular dimension measurements, including vascular stiffness and IMT determinations.
A multispectral scanner has been adapted to capture spectral images of living plants under various types of illumination for purposes of monitoring the health of, or monitoring the transfer of genes into, the plants. In a health-monitoring application, the plants are illuminated with full-spectrum visible and near infrared light and the scanner is used to acquire a reflected-light spectral signature known to be indicative of the health of the plants. In a gene-transfer- monitoring application, the plants are illuminated with blue or ultraviolet light and the scanner is used to capture fluorescence images from a green fluorescent protein (GFP) that is expressed as result of the gene transfer. The choice of wavelength of the illumination and the wavelength of the fluorescence to be monitored depends on the specific GFP.
Studying exogenously expressed tagged proteins in live cells has become a standard technique for evaluating protein distribution and function. Typically, expression levels of experimentally introduced proteins are not regulated, and high levels are often preferred to facilitate detection. However, overexpression of many proteins leads to mislocalization and pathologies. Therefore, for normative studies, moderate levels of expression may be more suitable. To understand better the dynamics of intermediate filament formation, transport, and stability in a healthy, living cell, we inserted neurofilament heavy chain (NFH)-green fluorescent protein (GFP) fusion constructs in adenoviral vectors with tetracycline (tet)-regulated promoters. This system allows for turning on or off the synthesis of NFH-GFP at a selected time, for a defined period, in a dose-dependent manner. We used this inducible system for live cell imaging of changes in filament structure and cell shape, motility, and transport associated with increasing NFH-GFP expression. Cells with low to intermediate levels of NFH-GFP were structurally and functionally similar to neighboring, nonexpressing cells. In contrast, overexpression led to pathological alterations in both filament organization and cell function. Copyright 2002 Wiley-Liss, Inc.
A unique subfamily of calmodulin-dependent Ca2+-ATPases was recently identified in plants. In contrast to the most closely related pumps in animals, plasma membrane-type Ca2+-ATPases, members of this new subfamily are distinguished by a calmodulin-regulated autoinhibitor located at the N-terminal instead of a C-terminal end. In addition, at least some isoforms appear to reside in non-plasma membrane locations. To begin delineating their functions, we investigated the subcellular localization of isoform ACA2p (Arabidopsis Ca2+-ATPase, isoform 2 protein) in Arabidopsis. Here we provide evidence that ACA2p resides in the endoplasmic reticulum (ER). In buoyant density sucrose gradients performed with and without Mg2+, ACA2p cofractionated with an ER membrane marker and a typical "ER-type" Ca2+-ATPase, ACA3p/ECA1p. To visualize its subcellular localization, ACA2p was tagged with a green fluorescence protein at its C terminus (ACA2-GFPp) and expressed in transgenic Arabidopsis. We collected fluorescence images from live root cells using confocal and computational optical-sectioning microscopy. ACA2-GFPp appeared as a fluorescent reticulum, consistent with an ER location. In addition, we observed strong fluorescence around the nuclei of mature epidermal cells, which is consistent with the hypothesis that ACA2p may also function in the nuclear envelope. An ER location makes ACA2p distinct from all other calmodulin-regulated pumps identified in plants or animals.
We developed a biochemical kinetics approach to describe the repair of double strand breaks (DSB) produced by low LET radiation by modeling molecular events associated with the mechanisms of non-homologous end-joining (NHEJ). A system of coupled non-linear ordinary differential equations describes the induction of DSB and activation pathways for major NHEJ components including Ku(sub 70/80), DNA-PK(sub cs), and the Ligase IV-XRCC4 hetero-dimer. The autophosphorylation of DNA-PK(sub cs and subsequent induction of gamma-H2AX foci observed after ionizing radiation exposure were modeled. A two-step model of DNA-PK(sub cs) regulation of repair was developed with the initial step allowing access of other NHEJ components to breaks, and a second step limiting access to Ligase IV-XRCC4. Our model assumes that the transition from the first to second-step depends on DSB complexity, with a much slower-rate for complex DSB. The model faithfully reproduced several experimental data sets, including DSB rejoining as measured by pulsed-field electrophoresis (PFGE), quantification of the induction of gamma-H2AX foci, and live cell imaging of the induction of Ku(sub 70/80). Predictions are made for the behaviors of NHEJ components at low doses and dose-rates, where a steady-state is found at dose-rates of 0.1 Gy/hr or lower.
The NASA-ISRO Synthetic Aperture Radar (NISAR) mission will carry L-band and S-band SAR instruments, each with a 240 km swath width. The L-band instrument has the capability to map the entire Earth’s land and ice covered surfaces from both ascending and descending orbit positions, continuously in an exact 12 day repeating cycle, given temporally dense (better than 6-day on average) sampling of Earth over the life of the mission. To achieve this swath coverage without loss of resolution or polarimetric capability, NISAR uses a reflector-feed based antenna system with scan-on-receive (“SweepSAR”) capability. One of the characteristics of SweepSAR is that the pulse repetition interval of the radar is shorter than the echo receive window for the 240 km swath. Therefore, transmit events occur during the receive window and the receivers must be blanked periodically, creating gaps in the coverage. For fixed pulse rate operations, these gaps are persistent strips of blanked ranges. NISAR is being designed to allow variation of the pulse rate in order to spread out these gaps throughout the synthetic aperture, but processing these variably-acquired data is more challenging and can lead to compromises in image quality. At the highest level NISAR places requirements on science measurements rather than image quality, so there is often a debate among science and engineering team members as to whether to operate with fixed pulse rates – leading to range strips consistently blank from cycle to cycle but with optimal image quality – or to vary the pulse rate, fill in the gaps, and live with degraded image quality. NISAR’s performance team has shown that science requirements can be met with the gaps. However, scientists are eager for maximum and coverage within a swath. A companion paper (Villano et al.) describes image quality for NISAR using a variable PRF approach, but does not go further to science requirements. This paper explores the issues when gaps are present in the swath due to fixed PRF operations.
Although not available to all patients with narrowed arteries, balloon angioplasty has expanded dramatically since its introduction with an estimated further growth to 562,000 procedures in the U.S. alone by 1992. Growth has fueled demand for higher quality imaging systems that allow the cardiologist to be more accurate and increase the chances of a successful procedure. A major advance is the Digital Cardiac Imaging (DCI) System designed by Philips Medical Systems International, Best, The Netherlands and marketed in the U.S. by Philips Medical Systems North America Company. The key benefit is significantly improved real-time imaging and the ability to employ image enhancement techniques to bring out added details. Using a cordless control unit, the cardiologist can manipulate images to make immediate assessment, compare live x-ray and roadmap images by placing them side-by-side on monitor screens, or compare pre-procedure and post procedure conditions. The Philips DCI improves the cardiologist's precision by expanding the information available to him.
Gravity-induced root curvature has long been considered to be regulated by differential distribution of the plant hormone auxin. However, the cells establishing these gradients, and the transport mechanisms involved, remain to be identified. Here, we describe a GFP-based auxin biosensor to monitor auxin during Arabidopsis root gravitropism at cellular resolution. We identify elevated auxin levels at the root apex in columella cells, the site of gravity perception, and an asymmetric auxin flux from these cells to the lateral root cap (LRC) and toward the elongation zone after gravistimulation. We differentiate between an efflux-dependent lateral auxin transport from columella to LRC cells, and an efflux- and influx-dependent basipetal transport from the LRC to the elongation zone. We further demonstrate that endogenous gravitropic auxin gradients develop even in the presence of an exogenous source of auxin. Live-cell auxin imaging provides unprecedented insights into gravity-regulated auxin flux at cellular resolution, and strongly suggests that this flux is a prerequisite for root gravitropism.
During the past two decades the use of live cytoskeletal probes has increased dramatically due to the introduction of the green fluorescent protein. However, to make full use of these live cell reporters it is necessary to implement simple methods to maintain plant specimens in optimal growing conditions during imaging. To image the cytoskeleton in living Arabidopsis roots, we rely on a system involving coverslips coated with nutrient supplemented agar where the seeds are directly germinated. This coverslip system can be conveniently transferred to the stage of a confocal microscope with minimal disturbance to the growth of the seedling. For roots with a larger diameter such as Medicago truncatula, seeds are first germinated in moist paper, grown vertically in between plastic trays, and roots mounted on glass slides for confocal imaging. Parallel with our live cell imaging approaches, we routinely process fixed plant material via indirect immunofluorescence. For these methods we typically use non-embedded vibratome-sectioned and whole mount permeabilized root tissue. The clearly defined developmental regions of the root provide us with an elegant system to further understand the cytoskeletal basis of plant development.
Tracker is an object-tracking and image-processing program designed and developed at the NASA Lewis Research Center to help with the analysis of images generated by microgravity combustion and fluid physics experiments. Experiments are often recorded on film or videotape for analysis later. Tracker automates the process of examining each frame of the recorded experiment, performing image-processing operations to bring out the desired detail, and recording the positions of the objects of interest. It can load sequences of images from disk files or acquire images (via a frame grabber) from film transports, videotape, laser disks, or a live camera. Tracker controls the image source to automatically advance to the next frame. It can employ a large array of image-processing operations to enhance the detail of the acquired images and can analyze an arbitrarily large number of objects simultaneously. Several different tracking algorithms are available, including conventional threshold and correlation-based techniques, and more esoteric procedures such as "snake" tracking and automated recognition of character data in the image. The Tracker software was written to be operated by researchers, thus every attempt was made to make the software as user friendly and self-explanatory as possible. Tracker is used by most of the microgravity combustion and fluid physics experiments performed by Lewis, and by visiting researchers. This includes experiments performed on the space shuttles, Mir, sounding rockets, zero-g research airplanes, drop towers, and ground-based laboratories. This software automates the analysis of the flame or liquid s physical parameters such as position, velocity, acceleration, size, shape, intensity characteristics, color, and centroid, as well as a number of other measurements. It can perform these operations on multiple objects simultaneously. Another key feature of Tracker is that it performs optical character recognition (OCR). This feature is useful in extracting numerical instrumentation data that are embedded in images. All the results are saved in files for further data reduction and graphing. There are currently three Tracking Systems (workstations) operating near the laboratories and offices of Lewis Microgravity Science Division researchers. These systems are used independently by students, scientists, and university-based principal investigators. The researchers bring their tapes or films to the workstation and perform the tracking analysis. The resultant data files generated by the tracking process can then be analyzed on the spot, although most of the time researchers prefer to transfer them via the network to their offices for further analysis or plotting. In addition, many researchers have installed Tracker on computers in their office for desktop analysis of digital image sequences, which can be digitized by the Tracking System or some other means. Tracker has not only provided a capability to efficiently and automatically analyze large volumes of data, saving many hours of tedious work, but has also provided new capabilities to extract valuable information and phenomena that was heretofore undetected and unexploited.
We examine Saturn's atmospheric dynamics with observations in the visible range from ground-based telescopes and Hubble Space Telescope (HST). We present a detailed analysis of observations acquired during 2018 obtaining drift rates of major meteorological systems from the equator to the North polar hexagon. A system of polar storms that appeared in the planet in March 2018 and remained active with a complex phenomenology at least until September is analyzed elsewhere [Sanchez-Lavega et al., 2019]. Many of the regular cloud features visible in 2018 are long-lived and can be identified in Saturn images in 2017, and in some cases, for up to a decade using also Cassini ISS images. Without considering the polar storms, the most interesting long-lived cloud systems are: i) A bright white spot in the Equatorial Zone that can be tracked continuously since 2014 with minimal changes in its zonal velocity, which was 444:3 +or- 3:1 ms(exp -1) in 2014 and 452:4 + or - 1:7 ms(exp -1) in 2018. This velocity is remarkably different from the zonal winds at the cloud level at its latitude during the Cassini mission, and is closer to zonal winds obtained at the time of the Voyagers flybys and to zonal winds from Cassini VIMS infrared images of the lower atmosphere. ii) A large long-lived Anticyclone Vortex, here AV, that formed after the Great White Spot of 2010-2011. This vortex has changed significantly in visual contrast, drift rate and latitude with minor changes in size over the last years. iii) A system of subpolar vortices at latitudes 60-65N present at least since 2011. These vortices and additional atmospheric features here studied follow drift rates consistent with zonal winds obtained by Cassini. We also present a study of the positions of the vertices of Saturn's North polar hexagon from 2015 to 2018. These measurements are compared with previous analyses during the Cassini mission (2007-2014), observations with HST in the 90s, and data from the Voyagers in 1980-1981 to explore the long term variability of the hexagon's drift rate. We find variations in the drift rate of the hexagon through these epochs that can not be fit by seasonal changes in the polar area. Instead, the different drift rates reinforce the role of the North Polar Spot that was present in the Voyager epoch and in the early 90s to cause a faster drift rate of the hexagon at that time compared with the current slower one.
Compact, reliable, rugged, automated cell-culture and frequency-agile microspectrofluorimetric apparatus developed to perform experiments involving photometric imaging observations of single live cells. In original application, apparatus operates mostly unattended aboard spacecraft; potential terrestrial applications include automated or semiautomated diagnosis of pathological tissues in clinical laboratories, biomedical instrumentation, monitoring of biological process streams, and portable instrumentation for testing biological conditions in various environments. Offers obvious advantages over present laboratory instrumentation.
Echocardiography is ideally suited for cardiovascular imaging in remote environments, but the expertise to perform it is often lacking. In 2001, an ATL HDI5000 was delivered to the International Space Station (ISS). The instrument is currently being used in a study to investigate the impact of long-term microgravity on cardiovascular function. The purpose of this report is to describe the methodology for remote guidance of echocardiography in space. Methods: In the year before launch of an ISS mission, potential astronaut echocardiographic operators participate in 5 sessions to train for echo acquisitions that occur roughly monthly during the mission, including one exercise echocardiogram. The focus of training is familiarity with the study protocol and remote guidance procedures. On-orbit, real-time guidance of in-flight acquisitions is provided by a sonographer in the Telescience Center of Mission Control. Physician investigators with remote access are able to relay comments on image optimization to the sonographer. Live video feed is relayed from the ISS to the ground via the Tracking and Data Relay Satellite System with a 2 second transmission delay. The expert sonographer uses these images along with two-way audio to provide instructions and feedback. Images are stored in non-compressed DICOM format for asynchronous relay to the ground for subsequent off-line analysis. Results: Since June, 2009, a total of 19 resting echocardiograms and 4 exercise studies have been performed in-flight. Average acquisition time has been 45 minutes, reflecting 26,000 km of ISS travel per study. Image quality has been adequate in all studies, but remote guidance has proven imperative for fine-tuning imaging and prioritizing views when communication outages limit the study duration. Typical resting studies have included 12 video loops and 21 still-frame images requiring 750 MB of storage. Conclusions: Despite limited crew training, remote guidance allows research-quality echocardiography to be performed by non-experts aboard the ISS. Analysis is underway and additional subjects are being recruited to define the impact of microgravity on cardiac structure and systolic and diastolic function.