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High Temporal Resolution (high + temporal_resolution)
Selected AbstractsBall-Pen Probe Measurements in L-Mode and H-Mode on ASDEX UpgradeCONTRIBUTIONS TO PLASMA PHYSICS, Issue 9 2010J. Adamek Abstract Experimental investigations of the plasma potential, poloidal electric field and electron temperature during L-mode and ELMy H-mode were performed on ASDEX Upgrade by means of a probe head containing four ball-pen probes and four Langmuir probes. This allows to measure simultaneously the floating and plasma potential at the same time which are related by the electron temperature. Thus a combination of ball-pen probes and Langmuir probes offers the possibility to determine the electron temperature directly with high temporal resolution. This novel temperature measurement method is compared to standard techniques. The influence of the electron temperature on the usual calculation of the poloidal electric field from the gradient of the floating potential is determined by a comparison to the poloidal electric field derived from the plasma potential (© 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] High speed sliding of axonemal microtubules produced by outer arm dyneinCYTOSKELETON, Issue 2 2005Raviraja N. Seetharam Abstract To study dynein arm activity at high temporal resolution, axonemal sliding was measured field by field for wild type and dynein arm mutants of Tetrahymena thermophila. For wt SB255 cells, when the rate of data acquisition was 60 fps, about 5× greater than previously published observations, sliding was observed to be discontinuous with very high velocity sliding (average 196 ,m/sec) for a few msec (1 or 2 fields) followed by a pause of several fields. The sliding velocities measured were an order of magnitude greater than rates previously measured by video analysis. However, when the data were analyzed at 12 fps for the same axonemes, consistent with previous observations, sliding was linear as the axonemes extended several times their original length with an average velocity of ,10 ,m/sec. The pauses or stops occurred at approximately 200 and 300% of the initial length, suggesting that dynein arms on one axonemal doublet were initially active to the limit of extension, and then the arms on the next doublet became activated. In contrast, in a mutant where OADs are missing, sliding observed at 60 fps was continuous and slow (5 ,m/sec), as opposed to the discontinuous high-velocity sliding of SB255 and of the mutant at the permissive temperature where OADs are present. High-velocity step-wise sliding was also present in axonemes from an inner arm dynein mutant (KO6). These results indicate that the high-speed discontinuous pattern of sliding is produced by the mechanochemical activity of outer arm dynein. The rate of sliding is consistent with a low duty ratio of the outer arm dynein and with the operation of each arm along a doublet once per beat. Cell Motil. Cytoskeleton 60:96,103, 2005. © 2004 Wiley-Liss, Inc. [source] Measurement of barbed ends, actin polymerization, and motility in live carcinoma cells after growth factor stimulation,CYTOSKELETON, Issue 4 2004Mike Lorenz Abstract Motility is associated with the ability to extend F-actin-rich protrusions and depends on free barbed ends as new actin polymerization sites. To understand the function and regulation of different proteins involved in the process of generating barbed ends, e.g., cofilin and Arp2/3, fixed cell approaches have been used to determine the relative barbed end concentration in cells. The major disadvantages of these approaches are permeabilization and fixation of cells. In this work, we describe a new live-cell time-lapse microscopy assay to determine the increase of barbed ends after cell stimulation that does not use permeabilization and provides a better time resolution. We established a metastatic carcinoma cell line (MTLn3) stably expressing GFP-,-actin at physiological levels. Stimulation of MTLn3 cells with epidermal growth factor (EGF) causes rapid and transient lamellipod protrusion along with an increase in actin polymerization at the leading edge, which can be followed in live cell experiments. By measuring the increase of F-actin at the leading edge vs. time, we were able to determine the relative increase of barbed ends after stimulation with a high temporal resolution. The F-actin as well as the barbed end concentration agrees well with published data for this cell line. Using this newly developed assay, a decrease in lamellipod extension and a large reduction of barbed ends was documented after microinjecting an anti-cofilin function blocking antibody. This assay has a high potential for applications where rapid changes in the dynamic filament population are to be measured. Cell Motil. Cytoskeleton 57:207,217, 2004. © 2004 Wiley-Liss, Inc. [source] Event-related delta oscillatory responses of Alzheimer patientsEUROPEAN JOURNAL OF NEUROLOGY, Issue 6 2008G. Yener Background and purpose:, Alzheimer type of dementia (AD) is the most common neuropsychiatric morbidity in elderly individuals. Event-related oscillations (ERO) provide an useful tool for detecting subtle abnormalities of cognitive processes with high temporal resolution. Methods:, In the present report, event-related oscillations of patients with AD were analyzed by using a visual oddball paradigm. A total of 22 mild probable AD subjects according to NINCDS-ADRDA criteria and 20 age-, gender-, and education-matched healthy control subjects were compared. AD group consisted from 11 untreated patients and 11 patients treated with cholinesterase inhibitor. Oscillatory responses were recorded from 13 scalp electrodes. Results:, Significant differences in delta frequency range were seen between the groups by using repeated measures of anova analysis [F(9.120) = 2.228; P = 0.022]. Post-hoc analyses using Wilcoxon test showed that at mid- and left central regions, (Cz, C3) peak amplitudes of delta responses of healthy subjects were significantly higher than either group. Also cholinesterase inhibitors did not have effect on delta oscillatory responses. Conclusions:, Our findings imply that the delta oscillatory responses at central locations are highly instable in mild probable AD patients regardless of treatment when compared to the healthy aged controls. This study supports the importance of oscillatory event-related potentials for investigating AD brain dynamics. [source] Drifting grating stimulation reveals particular activation properties of visual neurons in the caudate nucleusEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 7 2008Attila Nagy Abstract The role of the caudate nucleus (CN) in motor control has been widely studied. Less attention has been paid to the dynamics of visual feedback in motor actions, which is a relevant function of the basal ganglia during the control of eye and body movements. We therefore set out to analyse the visual information processing of neurons in the feline CN. Extracellular single-unit recordings were performed in the CN, where the neuronal responses to drifting gratings of various spatial and temporal frequencies were recorded. The responses of the CN neurons were modulated by the temporal frequency of the grating. The CN units responded optimally to gratings of low spatial frequencies and exhibited low spatial resolution and fine spatial frequency tuning. By contrast, the CN neurons preferred high temporal frequencies, and exhibited high temporal resolution and fine temporal frequency tuning. The spatial and temporal visual properties of the CN neurons enable them to act as spatiotemporal filters. These properties are similar to those observed in certain feline extrageniculate visual structures, i.e. in the superior colliculus, the suprageniculate nucleus and the anterior ectosylvian cortex, but differ strongly from those of the primary visual cortex and the lateral geniculate nucleus. Accordingly, our results suggest a functional relationship of the CN to the extrageniculate tecto-thalamo-cortical system. This system of the mammalian brain may be involved in motion detection, especially in velocity analysis of moving objects, facilitating the detection of changes during the animal's movement. [source] Visualization of local Ca2+ dynamics with genetically encoded bioluminescent reportersEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 3 2005Kelly L. Rogers Abstract Measurements of local Ca2+ signalling at different developmental stages and/or in specific cell types is important for understanding aspects of brain functioning. The use of light excitation in fluorescence imaging can cause phototoxicity, photobleaching and auto-fluorescence. In contrast, bioluminescence does not require the input of radiative energy and can therefore be measured over long periods, with very high temporal resolution. Aequorin is a genetically encoded Ca2+ -sensitive bioluminescent protein, however, its low quantum yield prevents dynamic measurements of Ca2+ responses in single cells. To overcome this limitation, we recently reported the bi-functional Ca2+ reporter gene, GFP-aequorin (GA), which was developed specifically to improve the light output and stability of aequorin chimeras [V. Baubet, et al., (2000) PNAS, 97, 7260,7265]. In the current study, we have genetically targeted GA to different microdomains important in synaptic transmission, including to the mitochondrial matrix, endoplasmic reticulum, synaptic vesicles and to the postsynaptic density. We demonstrate that these reporters enable ,real-time' measurements of subcellular Ca2+ changes in single mammalian neurons using bioluminescence. The high signal-to-noise ratio of these reporters is also important in that it affords the visualization of Ca2+ dynamics in cell,cell communication in neuronal cultures and tissue slices. Further, we demonstrate the utility of this approach in ex-vivo preparations of mammalian retina, a paradigm in which external light input should be controlled. This represents a novel molecular imaging approach for non-invasive monitoring of local Ca2+ dynamics and cellular communication in tissue or whole animal studies. [source] Mechanisms of time-based figure,ground segregationEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 10 2003Farid I. Kandil Abstract Figure,ground segregation can rely on purely temporal information, that is, on short temporal delays between positional changes of elements in figure and ground (Kandil, F.I. & Fahle, M. (2001) Eur. J. Neurosci., 13, 2004,2008). Here, we investigate the underlying mechanisms by measuring temporal segregation thresholds for various kinds of motion cues. Segregation can rely on monocular first-order motion (based on luminance modulation) and second-order motion cues (contrast modulation) with a high temporal resolution of ,,20 ms. The mechanism can also use isoluminant motion with a reduced temporal resolution of 60 ms. Figure,ground segregation can be achieved even at presentation frequencies too high for human subjects to inspect successive frames individually. In contrast, when stimuli are presented dichoptically, i.e. separately to both eyes, subjects are unable to perceive any segregation, irrespective of temporal frequency. We propose that segregation in these displays is detected by a mechanism consisting of at least two stages. On the first level, standard motion or flicker detectors signal local positional changes (flips). On the second level, a segregation mechanism combines the local activities of the low-level detectors with high temporal precision. Our findings suggest that the segregation mechanism can rely on monocular detectors but not on binocular mechanisms. Moreover, the results oppose the idea that segregation in these displays is achieved by motion detectors of a higher order (motion-from-motion), but favour mechanisms sensitive to short temporal delays even without activation of higher-order motion detectors. [source] Glycolytic activation at the onset of contractions in isolated Xenopus laevis single myofibresEXPERIMENTAL PHYSIOLOGY, Issue 9 2008Brandon Walsh Intracellular pH (pHi) was measured in isolated Xenopus laevis single myofibres at the onset of contractions, with and without glycolytic blockade, to investigate the time course of glycolytic activation. Single myofibres (n= 8; CON) were incubated in 2,,7,-bis(carboxyethyl)-5(6)-carboxyfluorescein acetoyxmethyl ester (10 ,m; for fluorescence measurement of pHi) and stimulated for 15 s at 0.67 Hz in anoxia in the absence (control condition; CON) and presence of a glycolytic inhibitor (1 mm iodoacetic acid; IAA). Intracellular pHi and tension were continuously recorded, and the differences in pHi between conditions were used to estimate the activation time of glycolysis. An immediate and steady increase in pHi (initial alkalosis) at the onset of contractions was similar between CON and IAA trials for the first 9 s of the contractile bout. However, from six contractions (,10 s) throughout the remainder of the bout, IAA demonstrated a continued rise in pHi, in contrast to a progressive decrease in pHi in CON (P < 0.05). These results demonstrate, with high temporal resolution, that glycolysis is activated within six contractions (10 s at 0.67 Hz) in single Xenopus skeletal muscle fibres. [source] The timing of cognitive control in partially incongruent categorizationHUMAN BRAIN MAPPING, Issue 9 2008Antao Chen Abstract We designed a novel task, partially incongruent categorization (PIC), to examine the timing of cognitive control. In the PIC task, participants categorized the probe stimulus according to a specific concept, and the number of features corresponding to the concept was varied. When there was one feature (c1 condition), the probe would elicit only categorization, but when there was more than one feature (c2 and c3 conditions), the probe would also elicit cognitive control. Here, the high temporal resolution of event-related potentials (ERPs) was utilized to investigate the temporal patterns of activity during conflict detection and control. Cognitive control elicited a N2 that was much larger in response to c2 and c3 than c1 in stimulus-locked waveforms, and no difference was evident between c2 and c3. The N2 was followed by a P3 that was much less on c2 and c3 than c1 trials, with no difference between c2 and c3. A dipole source analysis for two difference waves, c2,c1 and c3,c1, further showed that the corresponding dipoles of the N2 and P3 in the cognitive control conditions were in the anterior cingulate cortex (ACC) and prefrontal cortex (PFC), respectively. Taken together, the present findings support that ERP components in response to the PIC task reflect the time course of cognitive control: the N2 responds to conflict information and subsequently activates the P3 to control this conflict. The connection between the ACC and PFC is supported by their sequential activation within trials. Hum Brain Mapp, 2008. © 2007 Wiley-Liss, Inc. [source] The surface radiation budget over North America: gridded data assessment and evaluation of regional climate modelsINTERNATIONAL JOURNAL OF CLIMATOLOGY, Issue 15 2009Marko Markovic Abstract While surface station observations of downwelling radiation offer accuracy at high temporal resolution, they do not easily allow an evaluation of model surface radiation budgets (SRB) over a wide geographical area. We evaluate three gridded SRB data sets against detailed observations from six surface radiation sites from the US surface radiation (SURFRAD) network. We subsequently use the most accurate surrogate observational data set for evaluation of model-simulated SRB. The data sets assessed are: ERA40,reanalysis of European Centre for Medium-Range Weather Forecasts (ECMWF), North American Regional Reanalysis (NARR),regional reanalysis of National Centres for Environmental Prediction (NCEP) and the surface radiative budget (SRB) from the International Satellite Cloud Climatology Project (ISCCP). Due to varying constraints with respect to temporal coverage of each data set, the evaluation period used in this study is 1996,2001, inclusive. The ERA40 downwelling longwave radiation (DLR) appears the most accurate surrogate observation, while both ERA40 and ISCCP show accurate results when the incoming shortwave radiation (ISR) is considered across the annual cycle. Winter DLR is less accurate in ISCCP with a positive bias and lack of very low (<200 Wm,2) flux values. The NARR SRB shows a large positive bias in the ISR throughout the annual cycle, linked to a significant underestimate of cloud cover. The ERA40 data are subsequently used to evaluate the simulated SRB in three regional climate models across North America. With respect to solar radiation, cloud cover biases are seen to be crucial, while for longwave fluxes both cloud fraction and in-cloud water content are important to simulate correctly. Inclusion of trace gases beyond H2O, CO2 and O3 appears necessary for an accurate calculation of clear-sky longwave radiation. Error compensation frequently occurs between the various components contributing to a model total-sky SRB. This is important to consider when trying to identify the underlying causes of errors in the simulated total SRB. Copyright © 2009 Royal Meteorological Society [source] High temporal resolution SSFP cine MRI for estimation of left ventricular diastolic parametersJOURNAL OF MAGNETIC RESONANCE IMAGING, Issue 4 2010Ramkumar Krishnamurthy MS Abstract Purpose: To obtain high temporal resolution (HTR) magnetic resonance (MR) steady-state free-precession (SSFP) cine cardiac images by using multichannel radiofrequency (RF) hardware and parallel imaging techniques; to study the effect of temporal resolution; and to compare the derived left ventricular (LV) diastolic filling parameters with echocardiographic results. Materials and Methods: HTR images were acquired in 13 healthy volunteers using a 1.5 T scanner with 32 RF channels and sensitivity encoding (SENSE) and k-t broad-use linear-acquisition speedup technique (k-t BLAST) imaging techniques. LV diastolic parameters were calculated and compared to conventional echocardiographic indices such as the isovolumic relaxation time (IVRT) and E/A ratio. The need for HTR was assessed and the MR results were compared with echocardiographic results. Results: The HTR (,6-ms) images yielded higher peak filling rates, peak ejection rates, and peak atrial filling rates. A progressive decline in filling and ejection rates was observed with worsening temporal resolution. The IVRTs and E/A ratios measured with MR versus echocardiography were in broad agreement. Also, SENSE and k-t BLAST yielded similar diastolic functional parameters. Conclusion: With SENSE or k-t BLAST and modern hardware, HTR cine images can be obtained. The lower temporal resolutions (30,50 ms) used in clinical practice reduce LV filling rates by ,30% and may hinder characterization of transient phenomena such as the IVRT. J. Magn. Reson. Imaging 2010;31:872,880. ©2010 Wiley-Liss, Inc. [source] CBF, BOLD, CBV, and CMRO2 fMRI signal temporal dynamics at 500-msec resolutionJOURNAL OF MAGNETIC RESONANCE IMAGING, Issue 3 2008Qiang Shen PhD Abstract Purpose To investigate the temporal dynamics of blood oxygenation level-dependent (BOLD), cerebral blood flow (CBF), cerebral blood volume (CBV), and cerebral metabolic rate of oxygen (CMRO2) changes due to forepaw stimulation with 500-msec resolution in a single setting. Materials and Methods Forepaw stimulation and hypercapnic challenge on rats were studied. CBF and BOLD functional MRI (fMRI) were measured using the pseudo-continuous arterial spin-labeling technique at 500-msec resolution. CBV fMRI was measured using monocrystalline iron-oxide particles following CBF and BOLD measurements in the same animals. CMRO2 change was estimated via the biophysical BOLD model with hypercapnic calibration. Percent changes and onset times were analyzed for the entire forepaw somatosensory cortices and three operationally defined cortical segments, denoted Layers I,III, IV,V, and VI. Results BOLD change was largest in Layers I,III, whereas CBF, CBV, and CMRO2 changes were largest in Layers IV,V. Among all fMRI signals in all layers, only the BOLD signal in Layers I,III showed a poststimulus undershoot. CBF and CBV dynamics were similar. Closer inspection showed that CBV increased slightly first (P < 0.05), but was slow to peak. CBF increased second, but peaked first. BOLD significantly lagged both CBF and CBV (P < 0.05). Conclusion This study provides important temporal dynamics of multiple fMRI signals at high temporal resolution in a single setting. J. Magn. Reson. Imaging 2008. © 2008 Wiley-Liss, Inc. [source] Real-time MRI of joint movement with trueFISPJOURNAL OF MAGNETIC RESONANCE IMAGING, Issue 6 2002Harald H. Quick MSc Abstract Purpose To develop a technique for dynamic magnetic resonance imaging (MRI) of joint motion based on a combination of real-time TrueFISP (fast imaging with steady state precession) imaging with surface radiofrequency (RF) coils. Materials and Methods The metacarpal, elbow, tarsal, and knee joint of five volunteers and the knees of four patients were examined with a real-time TrueFISP sequence during movement of the joints. Results All examined joints could be assessed under dynamic conditions with high image contrast and high temporal resolution. Conclusion Dynamic MRI of joints with TrueFISP is feasible and can provide information supplemental to static joint examinations. J. Magn. Reson. Imaging 2002;15:710,715. © 2002 Wiley-Liss, Inc. [source] Evaluation of rapid volume changes of substrate-adherent cells by conventional microscopy 3D imagingJOURNAL OF MICROSCOPY, Issue 3 2004F. BOUDREAULT Summary Precise measurement of rapid volume changes of substrate-adherent cells is essential to understand many aspects of cell physiology, yet techniques to evaluate volume changes with sufficient precision and high temporal resolution are limited. Here, we describe a novel imaging method that surveys the rapid morphology modifications of living, substrate-adherent cells based on phase-contrast, digital video microscopy. Cells grown on a glass substrate are mounted in a custom-designed, side-viewing chamber and subjected to hypotonic swelling. Side-view images of the rapidly swelling cell, and at the end of the assay, an image of the same cell viewed from a perpendicular direction through the substrate, are acquired. Based on these images, off-line reconstruction of 3D cell morphology is performed, which precisely measures cell volume, height and surface at different points during cell volume changes. Volume evaluations are comparable to those obtained by confocal laser scanning microscopy (,Volume , 14%), but our method has superior temporal resolution limited only by the time of single-image acquisition, typically ,100 ms. The advantages of using standard phase-contrast microscopy without the need for cell staining or intense illumination to monitor cell volume make this system a promising new tool to investigate the fundamentals of cell volume physiology. [source] Removing undersampling artifacts in DCE-MRI studies using independent components analysisMAGNETIC RESONANCE IN MEDICINE, Issue 4 2008A.L. Martel Abstract In breast MRI mammography both high temporal resolution and high spatial resolution have been shown to be important in improving specificity. Adaptive methods such as projection reconstruction time-resolved imaging of contrast kinetics (PR-TRICKS) allow images to be reconstructed at various temporal and spatial resolutions from the same data set. The main disadvantage is that the undersampling, which is necessary to produce high temporal resolution images, leads to the presence of streak artifacts in the images. We present a novel method of removing these artifacts using independent components analysis (ICA) and demonstrate that this results in a significant improvement in image quality for both simulation studies and for patient dynamic contrast-enhanced (DCE)-MRI images. We also investigate the effect of artifacts on two quantitative measures of contrast enhancement. Using simulation studies we demonstrate that streak artifacts lead to pronounced periodic oscillations in pixel concentration curves which, in turn, lead to increased errors and introduce bias into heuristic measurements. ICA filtering significantly reduces this bias and improves accuracy. Pharmacokinetic modeling was more robust and there was no evidence of bias due to the presence of streak artifacts. ICA filtering did not significantly reduce the errors in the estimated pharmacokinetic parameters; however, the chi-squared error was greatly reduced after ICA filtering. Magn Reson Med, 2008. © 2008 Wiley-Liss, Inc. [source] High temporal and spatial resolution 4D MRA using spiral data sampling and sliding window reconstructionMAGNETIC RESONANCE IN MEDICINE, Issue 1 2004He Zhu Abstract Contrast-enhanced magnetic resonance angiography (CE-MRA) requires high spatial resolution to demonstrate detailed vasculature and high temporal resolution to capture the contrast bolus. Sparse bright voxels in MRA permit substantial undersampling in MRI data acquisition, allowing simultaneous high temporal and spatial resolution. We developed a time-resolved 3D MRA technique using the efficient spiral sampling trajectory, and performed off-resonance corrections using inhomogeneity field maps. View sharing and sliding window reconstruction were utilized to generate high temporal resolution. High-resolution 3D angiograms were generated at 1,2 s per frame, with a 5,8 ml gadolinium dose, in patients with vascular disease. Magn Reson Med 52:14,18, 2004. © 2004 Wiley-Liss, Inc. [source] In Vivo Phosphorescence Imaging of pO2 Using Planar Oxygen SensorsMICROCIRCULATION, Issue 6 2005PHILIPP BABILAS ABSTRACT Objective: Oxygen-dependent quenching of luminescence of metal porphyrin complexes has been used to image the pO2 distribution over tumor and normal tissue. Methods: An experimental setup is described using a platinum(II),octaethyl,porphyrin immobilized in a polystyrene matrix as transparent planar sensor. Results: Sensitivity over a broad range is high at low pO2 values (± 0.2 mm Hg at 0 mm Hg; ± 1.5 mm Hg at 160 mm Hg pO2). Due to intrinsically referencing via lifetime encoding there was no modification of the sensor response in vivo in the dorsal skinfold chamber model with amelanotic melanoma (A-MEL-3) in awake hamsters when compared to the in vitro calibration. pO2 measurements over normal tissue (25.8 ± 5.1 mm Hg) and tumor tissue (9.2 ± 5.1 mm Hg) were in excellent agreement with previous results obtained in this model using a surface multiwire electrode. Conclusions: Using the presented method the surface pO2 distribution can be mapped with a high temporal resolution of approximately 100 ms and a spatial resolution of at least 25 , m. Moreover, the transparent sensor allows the simultaneous visualization of the underlying microvasculature. [source] Towards metabolic mapping of the human retinaMICROSCOPY RESEARCH AND TECHNIQUE, Issue 5 2007D. Schweitzer Abstract Functional alterations are first signs of a starting pathological process. A device that measures parameter for the characterization of the metabolism at the human eye-ground would be a helpful tool for early diagnostics in stages when alterations are yet reversible. Measurements of blood flow and of oxygen saturation are necessary but not sufficient. The new technique of auto-fluorescence lifetime measurement (FLIM) opens in combination with selected excitation and emission ranges the possibility for metabolic mapping. FLIM not only adds an additional discrimination parameter to distinguish different fluorophores but also resolves different quenching states of the same fluorophore. Because of its high sensitivity and high temporal resolution, its capability to resolve multi-exponential decay functions, and its easy combination with laser scanner ophthalmoscopy, multi-dimensional time-correlated single photon counting was used for fundus imaging. An optimized set up for in vivo lifetime measurements at the human eye-ground will be explained. In this, the fundus fluorescence is excited at 446 or 468 nm and the time-resolved autofluorescence is detected in two spectral ranges between 510 and 560 nm as well as between 560 and 700 nm simultaneously. Exciting the fundus at 446 nm, several fluorescence maxima of lifetime t1 were detected between 100 and 220 ps in lifetime histograms of 40° fundus images. In contrast, excitation at 468 nm results in a single maximum of lifetime t1 = 190 ± 16 ps. Several fundus layers contribute to the fluorescence intensity in the short-wave emission range 510,560 nm. In contrast, the fluorescence intensity in the long-wave emission range between 560 and 700 nm is dominated by the fluorescence of lipofuscin in the retinal pigment epithelium. Comparing the lateral distribution of parameters of a tri-exponential model function in lifetime images of the fundus with the layered anatomical fundus structure, the shortest component (t1 = 190 ps) originates from the retinal pigment epithelium and the second lifetime (t2 = 1,000 ps) from the neural retina. The lifetime t3 , 5.5 ns might be influenced by the long decay of the fluorescence in the crystalline lens. In vitro analysis of the spectral properties of expected fluorophores under the condition of the living eye lightens the interpretation of in vivo measurements. Taking into account the transmission of the ocular media, the excitation of NADH is unlikely at the fundus. Microsc. Res. Tech., 2007. © 2007 Wiley-Liss, Inc. [source] An Improved Algorithm for Satellite-derived UV Radiation,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 1 2003Sigrid Wuttke ABSTRACT The improved algorithm surface irradiance derived from a range of satellite-based sensors (SIDES) is presented in this article. It calculates various types of surface UV intensities, such as biologically weighted or unweighted UV spectra, integrated doses or irradiance at specific wavelengths, using data from satellite instruments. These surface UV data are mainly useful for environmental impact or process studies where high accuracy or a high temporal resolution is required. In contrast to several previous studies, SIDES has been validated with spectral measurements. By this method an averaging of positive or negative deviations over the complete wavelength range is avoided. This is especially important for UV wavelengths around 300 nm where biological effectiveness is highest. The results of SIDES deviate less than 7% from ground-based observations for wavelengths between 295 and 400 nm. In contrast, the corresponding deviations of the joint research center algorithm escalate for shorter wavelengths, reaching 35% at 295 nm. This large deviation is due to an inaccurate interpolation procedure that has been detected by spectral analysis. Thus, spectral validation is demonstrated to be an appropriate tool to detect weaknesses in such an algorithm and provides information essential for improvement. [source] Internalization of Aggregated Photosensitizers by Tumor Cells: Subcellular Time-resolved Fluorescence Spectroscopy on Derivatives of Pyropheophorbide-a Ethers and Chlorin e6 under Femtosecond One- and Two-photon Excitation,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 6 2002L. Kelbauskas ABSTRACT Amphiphilic sensitizers self-associate in aqueous environments and form aggregated species that exhibit no or only negligible photodynamic activity. However, amphiphilic photosensitizers number among the most potent agents of photodynamic therapy. The processes by which these sensitizers are internalized into tumor cells have yet to be fully elucidated and thus remain the subject of debate. In this study the uptake of photosensitizer aggregates into tumor cells was examined directly using subcellular time-resolved fluorescence spectroscopy with a high temporal resolution (20,30 ps) and high sensitivity (time-correlated single-photon counting). The investigations were performed on selected sensitizers that exhibit short fluorescence decay times (<50 ps) in aggregated form. Derivatives of pyropheophorbide-a ether and chlorin e6 with varying lipophilicity were used for the study. The characteristic fluorescence decay times and spectroscopic features of the sensitizer aggregates measured in aqueous solution also could be observed in A431 human endothelial carcinoma cells administered with these photosensitizers. This shows that tumor cells can internalize sensitizers in aggregated form. Uptake of aggregates and their monomerization inside cells were demonstrated directly for the first time by means of fluorescence lifetime imaging with a high temporal resolution. Internalization of the aggregates seems to be endocytosis mediated. The degree of their monomerization in tumor cells is strongly influenced by the lipophilicity of the compounds. [source] Fine-root respiration in a loblolly pine (Pinus taeda L.) forest exposed to elevated CO2 and N fertilizationPLANT CELL & ENVIRONMENT, Issue 11 2008JOHN E. DRAKE ABSTRACT Forest ecosystems release large amounts of carbon to the atmosphere from fine-root respiration (Rr), but the control of this flux and its temperature sensitivity (Q10) are poorly understood. We attempted to: (1) identify the factors limiting this flux using additions of glucose and an electron transport uncoupler (carbonyl cyanide m-chlorophenylhydrazone); and (2) improve yearly estimates of Rr by directly measuring its Q10in situ using temperature-controlled cuvettes buried around intact, attached roots. The proximal limits of Rr of loblolly pine (Pinus taeda L.) trees exposed to free-air CO2 enrichment (FACE) and N fertilization were seasonally variable; enzyme capacity limited Rr in the winter, and a combination of substrate supply and adenylate availability limited Rr in summer months. The limiting factors of Rr were not affected by elevated CO2 or N fertilization. Elevated CO2 increased annual stand-level Rr by 34% whereas the combination of elevated CO2 and N fertilization reduced Rr by 40%. Measurements of in situ Rr with high temporal resolution detected diel patterns that were correlated with canopy photosynthesis with a lag of 1 d or less as measured by eddy covariance, indicating a dynamic link between canopy photosynthesis and root respiration. These results suggest that Rr is coupled to daily canopy photosynthesis and increases with carbon allocation below ground. [source] Elucidation of structure,function relationships in the lung: contributions from hyperpolarized 3helium MRICLINICAL PHYSIOLOGY AND FUNCTIONAL IMAGING, Issue 6 2002Hans-Ulrich Kauczor Summary Magnetic resonance imaging (MRI) using hyperpolarized 3helium (He) gas as the source of signal provides new physiological insights into the structure,function relationships of the lung. Traditionally, lung morphology has been visualized by chest radiography and computed tomography, whereas lung function was assessed by using nuclear medicine. As all these techniques rely on ionizing radiation, MRI has some inherent advantages. 3He MRI is based on ,optical pumping' of the 3He gas which increases the nuclear spin polarization by four to five orders of magnitude translating into a massive gain in signal. Hyperpolarized 3He gas is administered as an inhaled ,contrast agent' and allows for selective visualization of airways and airspaces. Straightforward gas density images demonstrate the homogeneity of ventilation with high spatial resolution. In patients with lung diseases 3He MRI has shown a high sensitivity to depict ventilation defects. As 3He has some more exciting properties, a comprehensive four-step functional imaging protocol has been established. The dynamic distribution of ventilation during continuous breathing can be visualized after inhalation of a single breath of 3He gas using magnetic resonance (MR) sequences with high temporal resolution. Diffusion weighted 3He MRI provides a new measure for pulmonary microstructure because the degree of restriction of the Brownian motion of the 3He atoms reflects lung structure. Since the decay of 3He hyperpolarization is dependent on the ambient oxygen concentration, regional and temporal analysis of intrapulmonary pO2 becomes feasible. Thus, pulmonary perfusion, ventilation,/perfusion ratio and oxygen uptake can be indirectly assessed. Further research will determine the significance of the functional information with regard to physiology and patient management. [source] |