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Real-time Measurement (real-time + measurement)
Selected AbstractsReal-time measurement of nitric oxide in single mature mouse skeletal muscle fibres during contractionsTHE JOURNAL OF PHYSIOLOGY, Issue 1 2007Deborah Pye Nitric oxide (NO) is thought to play multiple roles in skeletal muscle including regulation of some adaptations to contractile activity, but appropriate methods for the analysis of intracellular NO activity are lacking. In this study we have examined the intracellular generation of NO in isolated single mature mouse skeletal muscle fibres at rest and following a period of contractile activity. Muscle fibres were isolated from the flexor digitorum brevis muscle of mice and intracellular NO production was visualized in real-time using the fluorescent NO probe 4-amino-5-methylamino-2,,7,-difluorofluorescein diacetate (DAF-FM DA). Some leakage of DAF-FM was apparent from fibres loaded with the probe, but they retained sufficient probe to respond to changes in intracellular NO following addition of the NO donor 3-(2-hydroxy-1-methyl-2-nitrosohydrazino)- N -methyl-1-propanamine (NOC-7) up to 30 min after loading. Electrically stimulated contractions in isolated fibres increased the rate of change in DAF-FM fluorescence by ,48% compared to non-stimulated fibres (P < 0.05) and the rate of change in DAF-FM fluorescence in the stimulated fibres returned to control values by 5 min after contractions. Treatment of isolated fibres with the NO synthase inhibitors NG -nitro- l -arginine methyl ester hydrochloride (l -NAME) or NG -monomethyl- l -arginine (l -NMMA) reduced the increase in DAF-FM fluorescence observed in response to contractions of untreated fibres. Treatment of fibres with the cell-permeable superoxide scavenger 4,5-dihydroxy-1,3-benzenedisulphonic acid (Tiron) also reduced the increase in fluorescence observed during contractions suggesting that superoxide, or more probably peroxynitrite, contributes to the fluorescence observed. Thus this technique can be used to examine NO generation in quiescent and contracting skeletal muscle fibres in real time, although peroxynitrite and other reactive nitrogen species may potentially contribute to the fluorescence values observed. [source] Real-time measurement of serotonin release and motility in guinea pig ileumTHE JOURNAL OF PHYSIOLOGY, Issue 2 2006Paul P. Bertrand Enterochromaffin (EC) cells are sensors that detect chemical or mechanical stimuli and respond with release of serotonin (5-HT). 5-HT activates local motor reflexes, but whether local motor reflexes also evoke 5-HT release is unknown. The aim of the present study was to establish the relationship between the release of 5-HT and the enteric neural circuits controlling the movements of the intestine. Recordings were made from full-thickness preparations of guinea pig ileum using electrochemical techniques with carbon fibre electrodes to measure local concentrations of 5-HT. The tension in the circular muscle (CM) and longitudinal muscle (LM) was recorded with force transducers. The release of 5-HT from the EC cells was detected selectively and the timing of the events quantified. Pressure-evoked peristalsis caused detectable 5-HT release only when the recording site was invaded by a ring of CM contraction. Spontaneous and stretch-evoked reflex contraction of the CM and LM occurred simultaneously with 5-HT release. Paralysis of the smooth muscle significantly reduced the stretch-evoked release. Muscarinic agonists evoked reflexes that were associated with increases in tension in CM and LM simultaneous with 5-HT release. Tetrodotoxin abolished the coordination between the CM contraction and 5-HT release but not the direct activation of the CM and EC cells by the agonists. In conclusion, the correlation between local motor reflexes and 5-HT release observed in the present study is caused primarily by the contraction of the smooth muscle and subsequent deformation of the mucosa. The EC cell is, thus, a site of convergence for mechanical forces that contribute to the release of 5-HT during motor reflexes. [source] Real-time measurement of protein leaching from micro-particulate larval fish feedsAQUACULTURE RESEARCH, Issue 16 2008Peter M Nicklason Abstract The small size and high surface area to volume ratio of larval fish feed presents challenges for nutrient retention in micro-particulate diets. A method for the accurate and rapid measurement of nutrient retention or loss from micro-particulate feed in water is needed to help develop micro-particulate feeds with good nutrient retention characteristics. The present study developed and validated an instrument method using fibre optic technology that measures protein leaching in real time. Larval fish feed particles of different sizes (100,500 ,m) and formulations were measured. Under consistent experimental conditions, a feed could be assayed for the rate of mass loss and the half-life or time of 50% total soluble mass loss. The results closely approximated natural decay models with coefficients of determination (r2) >0.95. The end result is a fast and accurate method to quantify and provide solid reference data for a feed formulation or particle size. Using this method allows different feeds to be compared and conclusions drawn for relative performance. [source] Real-time measurement of cytosolic free calcium concentration in DEM-treated HL-60 cells during static magnetic field exposure and activation by ATPBIOELECTROMAGNETICS, Issue 3 2009Camilla Rozanski Abstract This study investigated whether glutathione depletion affected the sensitivity of HL-60 cells to static magnetic fields. The effect of Diethylmaleate (DEM) on static magnetic field induced changes in cytosolic free calcium concentration ([Ca2+]c) was examined. Cells were loaded with a fluorescent dye and exposed to a uniform static magnetic field at a strength of 0 mT (sham) or 100 mT. [Ca2+]c was monitored during field and sham exposure using a ratiometric fluorescence spectroscopy system. Cells were activated by the addition of ATP. Metrics extracted from the [Ca2+]c time series included: average [Ca2+]c during the Pre-Field and Field Conditions, peak [Ca2+]c following ATP activation and the full width at half maximum (FWHM) of the peak ATP response. Comparison of each calcium metric between the sham and 100 mT experiments revealed the following results: average [Ca2+]c measured during the Field condition was 53,±,2 nM and 58,±,2 nM for sham and 100 mT groups, respectively. Average FWHM was 51,±,3 s and 54,±,3 s for sham and 100 mT groups, respectively. An effect of experimental order on the peak [Ca2+]c response to ATP in sham/sham experiments complicated the statistical analysis and did not allow pooling of the first and second order experiments. No statistically significant difference between the sham and 100 mT groups was observed for any of the calcium metrics. These data suggested that manipulation of free radical buffering capacity in HL-60 cells did not affect the sensitivity of the cells to a 100 mT static magnetic field. Bioelectromagnetics 30:213,221, 2009. © 2008 Wiley-Liss, Inc. [source] Influence of bone density on the cement fixation of femoral hip resurfacing componentsJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 8 2010Rudi G. Bitsch Abstract In clinical outcome studies, small component sizes, female gender, femoral shape, focal bone defects, bad bone quality, and biomechanics have been associated with failures of resurfacing arthroplasties. We used a well-established experimental setup and human bone specimens to analyze the effects of bone density on cement fixation of femoral hip resurfacing components. Thirty-one fresh frozen femora were prepared for resurfacing using the original instruments. ASRÔ resurfacing prostheses were implanted after dual-energy X-ray densitometer scans. Real-time measurements of pressure and temperature during implantation, analyses of cement penetration, and measurements of micro motions under torque application were performed. The associations of bone density and measurement data were examined calculating regression lines and multiple correlation coefficients; acceptability was tested with ANOVA. We found significant relations between bone density and micro motion, cement penetration, cement mantle thickness, cement pressure, and interface temperature. Mean bone density of the femora was 0.82,±,0.13,g/cm2, t- score was ,0.7,±,1.0, and mean micro motion between bone and femoral resurfacing component was 17.5,±,9.1,µm/Nm. The regression line between bone density and micro motion was equal to ,56.7,×, bone density,+,63.8, R,=,0.815 (p,<,0.001). Bone density scans are most helpful for patient selection in hip resurfacing, and a better bone quality leads to higher initial component stability. A sophisticated cementing technique is recommended to avoid vigorous impaction and incomplete seating, since increasing bone density also results in higher cement pressures, lower cement penetration, lower interface temperatures, and thicker cement mantles. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:986,991, 2010 [source] A chip-based miniaturized format for protein-expression profiling: The exploitation of comprehensively produced antibodiesELECTROPHORESIS, Issue 18 2006Hisashi Koga Dr. Abstract Numerous antibodies have been developed and validated in recent years, and show promise for use in novel functional protein assays. Such assays would be an alternative to pre-existing comprehensive assays, such as DNA microarrays. Antibody microarrays are thought to represent those functional protein assays. While a variety of attempts have been made to apply DNA microarray technology to antibody microarrays, a fully optimized protocol has not been established. We have been conducting a project to comprehensively produce antibodies against mouse KIAA ("KI" stands for "Kazusa DNA Research Institute" and "AA" are reference characters) proteins. Using our library of antibodies, we established a novel antibody microarray format that utilizes surface plasmon resonance (SPR) technology. A label-free real-time measurement of protein expression in crude cell lysates was achieved by direct readout of the bindings using SPR. Further refinement of the antibody microarray format enabled us to detect a smaller quantity of target proteins in the lysate without the bulk effect. In this review, we first summarize available antibody array formats and then describe the above-mentioned format utilizing updated SPR technology. [source] Automatic Delineation of Capture Zones for Pump and Treat Systems: A Case Study in Piedmont, ItalyGROUND WATER MONITORING & REMEDIATION, Issue 2 2010Tiziana Tosco The design of a pump and treat (P&T) system for the hydraulic control of a contaminated plume in a confined aquifer is presented here. Being the system designed for the emergency containment of a nonaqueous phase liquid plume, the evaluation of the system's short-term efficiency was considered an important issue. For this reason, both time-related and ultimate capture zones were defined. They were traced using the automatic protection area (APA) model, a capture-zone delineation tool based on a hybrid forward-backward particle tracking algorithm, that provides an automatic post-processing encirclement of capture zones. Two simple indexes are here proposed for the evaluation of the performance of the hydraulic barrier, that is, the efficacy and efficiency indexes, calculated from the capture areas provided by APA. The discharge rates of the wells were dimensioned applying the APA algorithm, maximizing efficacy and efficiency of the barrier. Results proved both visually (via plotting of capture zones) and numerically (via calculation of the indexes) that the P&T system can provide a complete capture of the contaminated area and minimizes the volume of extracted water. Consequently, the APA algorithm was proved to be a useful tool in capture zone delineation. As a future perspective, it could be coupled with the real-time measurement of pumping rates and water levels and be implemented as a part of a tuning tool for the management of the hydraulic barrier. [source] Experimental study of feasibility in kinetically-controlled reactive distillationAICHE JOURNAL, Issue 2 2005Madhura Chiplunkar Abstract Bifurcation studies predict limited ranges of feasibility for products in certain reactive distillations. These are closely related to the bifurcations in the singular points of dynamic models for simple reactive distillation (isobaric open evaporation with liquid phase reaction). A new dynamic model is described with constant vapor rate together with an experimental study for the reactive distillation of acetic acid with isopropanol to produce isopropyl acetate, catalyzed by Amberlyst-15 ion-exchange resin. An experimental apparatus with real-time measurement of liquid compositions based on Fourier transform infrared (FTIR) spectroscopy is described, and used to follow the composition dynamics at several initial conditions and Damköhler numbers (Da). The experimental results match model predictions that show four regions of behavior. For Da , 1, these show a stable node at acetic acid and several other fixed points as saddles. However, near Da , 2, both isopropanol and acetic acid are stable nodes and a quaternary singular point appears. The presence of two stable nodes requires the presence of a distillation boundary and, therefore, a limited feasibility for the bottom product compositions from continuous reactive distillation. For the reaction rates studied, the model predictions are closely consistent with the experimental findings, and are robust to variations in the vapor rate. These experiments are among the first to analyze the dynamics and feasibility in a kinetically-controlled reactive distillation and are consistent with previous studies for the reaction equilibrium limit, indicating the formation of a reactive azeotrope. © 2005 American Institute of Chemical Engineers AIChE J, 51: 464,479, 2005 [source] Evaluation of a Novel Real-Time Continuous Glucose-Monitoring System for Use in CatsJOURNAL OF VETERINARY INTERNAL MEDICINE, Issue 1 2010S. Moretti Background: The Guardian REAL-Time is a continuous glucose-monitoring system (CGMS) recently developed to provide instantaneous interstitial glucose concentrations; the system does not require a monitor being fixed to the animal. Hypothesis: The CGMS provides accurate and reproducible real-time readings of glucose concentration in cats. Animals: Thirty-two diabetic cats, 2 cats with suspected insulinoma, and 5 healthy cats. Methods: Prospective, observational study. CGMS accuracy was compared with a reference glucose meter at normal, high, and low blood glucose concentrations using error grid analysis. Reading variability of 2 simultaneously used CGMS was determined in diabetic cats by calculating correlation and percentage of concordance of paired data at different glycemic ranges. The time interval between increasing glycemia and a rise in interstitial fluid glucose measured by the CGMS was assessed in healthy cats receiving glucose IV; the time point of maximal increase in interstitial glucose concentrations was calculated. Results: The CGMS was 100, 96.1, and 91.0% accurate at normal, high, and low blood glucose concentrations. Measurements deviated from reference by ,12.7 ± 70.5 mg/dL at normal, ,12.1 ± 141.5 mg/dL at high, and ,1.9 ± 40.9 mg/dL at low glucose concentrations. Overall, paired CGMS readings correlated significantly (r= 0.95, P < .0001) and concordance was 95.7%. The median delay after IV administration of glucose to an increase in interstitial glucose was 11.4 minutes (range: 8.8,19.7 minutes). Conclusions and Clinical Importance: Although some readings substantially deviated from reference values, the CGMS yields reproducible results, is clinically accurate in cats with hyperglycemia and euglycemia, and is slightly less accurate if blood glucose concentrations are low. Rapidly increasing interstitial glucose after a glycemic rise suggests that the CGMS is suitable for real-time measurement under clinical conditions. [source] Expediting sustainable brownfields redevelopment by applying Triad using the membrane interface probeREMEDIATION, Issue 4 2010Maria D. Watt Redevelopment and reuse plans are often based upon an expedited delineation and remediation life cycle, since delayed reuse usually has economic consequences. It has also become increasingly important to utilize sustainable practices to achieve investigation and remediation goals. In this article, the Triad approach is used to expedite the delineation of a source area within a municipal landfill to complete the remedial effort prior to construction of an urban civic center. The Triad approach uses the three elements of systematic project planning, dynamic work strategy, and real-time measurement to expedite site characterization (Interstate Technology and Regulatory Council, 2003). In this article, the Triad sampling strategy consisted of two phases. The first phase included in situ screening of soil and groundwater using the membrane interface probe (MIP), and the second phase included confirmatory sampling via vertical profiles in the soil and groundwater. This study found that, using the MIP in a dynamic sampling strategy, a critical element of the Triad approach, combined with the proper placement of confirmatory samples, significantly reduced overall project cost and will expedite the site redevelopment. The use of the Triad approach also contributed to the integration of green and sustainable practices into the project. © 2010 Wiley Periodicals, Inc. [source] Release of ATP in the central nervous system during systemic inflammation: real-time measurement in the hypothalamus of conscious rabbitsTHE JOURNAL OF PHYSIOLOGY, Issue 1 2007Alexander V. Gourine Receptors for extracellular ATP (both ionotropic and metabotropic) are widely expressed in the CNS both in neurones and glia. ATP can modulate neuronal activity in many parts of the brain and contributes to the central nervous control of several physiological functions. Here we show that during the systemic inflammatory response the extracellular concentrations of ATP increase in the anterior hypothalamus and this has a profound effect on the development of the thermoregulatory febrile response. In conscious rabbits we measured ATP release in real time with novel amperometric biosensors and monitored a marked increase in the concentration of ATP (4.0 ± 0.7 ,m) in the anterior hypothalamus in response to intravenous injection of bacterial endotoxin , lipopolysaccharide (LPS). No ATP release was observed in the posterior hypothalamus. The release of ATP coincided with the development of the initial phase of the febrile response, starting 18 ± 2 min and reaching its peak 45 ± 2 min after LPS injection. Application of the ATP receptor antagonists pyridoxal-5,-phosphate-6-azophenyl-2,,4,-disulphonic acid, Brilliant Blue G or periodate oxidized ATP dialdehyde to the site of ATP release in the anterior hypothalamus markedly augmented and prolonged the febrile response. These data indicate that during the development of the systemic inflammation, ATP is released in the anterior hypothalamus to limit the magnitude and duration of fever. This release may also have a profound effect on the hypothalamic control of other physiological functions in which ATP and related purines have been implicated to play modulatory roles, such as food intake, hormone secretion, cardiovascular activity and sleep. [source] Estimation of the base line imagined as a muscular background for myoglobin determination by reflectance spectrometryANIMAL SCIENCE JOURNAL, Issue 6 2004Masatoshi IZUMIMOTO ABSTRACT Liquid extraction method for determination of myoglobin in muscle is not suitable because of changes in the derivatives and time required. Optical reflectance method is useful as a non-destructive and real-time measurement. However, the base line of reflectance method, the muscular reflectance background (MRB) imagined as colorless structural elements, has been unknown because of non-existence. The spectrum of the base line was analyzed as the analogy of bleached meat. A formula was derived for reflectance that had a linear relationship to the reciprocal of wavelength to the power 4. The reflectance characteristic of the MRB was not constant and the curve tended to rise with increasing wavelength. The reflectance at 700 nm was estimated to be 50%. By the characteristic analysis on Commission Internationale l'Eclairage color values and reciprocal reflectance, it was concluded that the reflectance spectrum of the muscular background obtained in this research was fitted as a base line for the reflectance spectrometry. [source] Venous Pressure Regulation During Pulsatile Extracorporeal Life SupportARTIFICIAL ORGANS, Issue 10 2008Seong Wook Choi Abstract During the use of pulsatile extracorporeal life support (pulsatile-ECLS), the patient's venous pressure near the inlet venous catheter tip must be monitored to maintain sufficient blood flow and to prevent vein collapse. However, direct measurement of the venous pressure and the estimate of suction using measured blood inflow and prepump pressure are not practical because of setup difficulties during emergency treatments and in cardiovascular operations. In this article, we describe a new method for estimating the venous pressure that can be implemented in the controller of the pulsatile-ECLS system, the T-PLS. It uses real-time measurement of the electric current and actuator motion. The current waveform of the T-PLS is used to determine the outflow amount and the volume remaining in the pulsatile pumps. Previously measured values of the pulsatile-ECLS compliance and the hemodynamic resistance along the inflow path are used to evaluate venous pressure with estimated blood flow. Estimated prepump pressure, inflow, and venous pressure were compared to the measured data in a series of in vitro experiments. The estimated venous pressure was used to avoid vein collapse and to increase the reliability in animal experiments. [source] State estimation of networks with distributed generationEUROPEAN TRANSACTIONS ON ELECTRICAL POWER, Issue 1 2007Iñigo Cobelo Abstract The nature and control of existing distribution networks limits the amount of distributed generation that can be connected. To increase the penetration of distributed generation a distribution management system controller (DMSC) can be used. The use of a DMSC requires a state estimator algorithm that provides an estimate of the network state in real time. In this paper, a state estimation algorithm based on the methods used on transmission networks is presented. The application of these methods into distribution networks requires existing measuring systems to be supplemented with the addition of new real-time measurements, and the use of load estimates. Copyright © 2006 John Wiley & Sons, Ltd. [source] An adaptive resource reservation for vehicular mobile networksINTERNATIONAL JOURNAL OF NETWORK MANAGEMENT, Issue 5 2009I. Ben Hamida This paper presents the time-based bandwidth reservation (TBR) algorithm, suitable for handoff management in cellular systems. TBR is based on real-time measurements of mobile stations (position, velocity and acceleration). The scheme consists in sending reservation requests to the neighboring cells based on an extrapolation of the user's motion. The originality of our approach lies in dynamically adjusting the amount of time for which bandwidth has to be allocated and reserved in a cell. In addition, we propose an optimal channel requests arrangement (CRA) algorithm in order to improve the performance of TBR in terms of resource utilization. Finally, we propose VTBR, an adapted and extended version of TBR for better support of vehicular network specificities where service degradation or forced call termination may occur owing to frequent handoffs. Detailed simulation results for TBR and VTBR schemes and a comparison with the guard channel scheme are presented. The results show that TBR and VTBR can efficiently improve the flow dropping probability. Copyright © 2008 John Wiley & Sons, Ltd. [source] Assessing arterial blood flow and vessel area variations using real-time zonal phase-contrast MRIJOURNAL OF MAGNETIC RESONANCE IMAGING, Issue 3 2006Markus Oelhafen PhD Abstract Purpose To measure peripheral artery function using a real-time phase-contrast (PC)-MRI sequence with tailored image-processing algorithms for flow computation. Materials and Methods An approach to real-time flow measurements was developed based on two-dimensional spatially selective excitation pulses and consecutive tailored processing of the data to derive blood flow and vessel area variations. The data acquisition strategy allows for flow measurements at high spatial and temporal resolutions of 1 mm2 and 50 msec, respectively. In postprocessing the vessel area is automatically extracted using correlation measures in conjunction with morphological image operators. By means of in vitro and in vivo validations, it is shown that the current methods provide accurate and reproducible measurements of flow and vessel area variations. Results In vitro the comparison between the lumen area measured with the presented method and the values obtained by calliper gauge measurement showed a difference of 3.4% ± 3.4% (mean ± 2 SD). Similarly, the comparison between the stroke volumes determined with the presented method and by stopwatch and bucket measurements yielded a difference of 6.1% ± 2.1%. In vivo the results from the real-time measurements for lumen area and stroke volume were compared with those from a gated PC-MRI technique with differences of 4.8% ± 14% and 3.0% ± 24.7%, respectively. Conclusion The presented method constitutes a reliable tool set for quantifying the variations of blood flow and lumen area in the superficial femoral artery during reactive hyperemia and for studying their correlation with cardiovascular risk factors. J. Magn. Reson. Imaging 2006. © 2006 Wiley-Liss, Inc. [source] EVIDENCE OF A LATENT OXIDATIVE BURST IN RELATION TO WOUND REPAIR IN THE GIANT UNICELLULAR CHLOROPHYTE DASYCLADUS VERMICULARIS,JOURNAL OF PHYCOLOGY, Issue 3 2005Cliff Ross We investigated the kinetics and composition of the second phase of the wound repair process of Dasycladus vermicularis ([Scropoli] Krasser) using fluorescent probes, chromatography, UV spectroscopy, and histochemistry. Our new evidence supports the hypothesis that the second phase of wound repair (initiated at approximately 35,45 min postinjury) is based on the activation of an oxidative burst that produces micromolar H2O2 levels. These results provide evidence of peroxidase activity at the wound site, real-time measurements of an oxidative burst, and catechol localization in wound plugs. Strong evidence is presented indicating that the biochemical machinery exists for oxidative cross-linking to ensue in the wound-healing process of D. vermicularis. [source] Systematic planning for Triad projectsREMEDIATION, Issue 1 2004Robert Howe This article examines specific systematic planning steps that can be used for designing and controlling Triad projects. Triad work strategies act to limit decision uncertainty, expedite schedules to meet project milestones, and reduce costs associated with cleanup activities. As a result, the Triad approach is rapidly increasing in popularity. Good project planning has always been seen as the cornerstone of successful Triad projects. However, the specific steps in the systematic planning process have not been extensively published. Demands of Triad projects, which attempt to make maximum use of innovative technologies and sequencing of activities in a learn- as-you-go framework, put new demands on regulators and project managers alike. Specific activities and relationships are identified to assist project managers with dynamic work strategies and real-time measurements to support improved decision making. These include: assembly of stakeholders, a core technical team, and key decisions; development and refinement of a site model; use of demonstrations of methods applicability; development of dynamic work strategies and project sequencing; real-time data management assessment and presentation; and unitized procurement of technologies and services. © 2004 Wiley Periodicals, Inc. [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] |