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1.
Object tracking is an instrumental tool supporting studies of cellular trafficking. There are three challenges in object tracking: the identification of targets; the precise determination of their position and boundaries; and the assembly of correct trajectories. This last challenge is particularly relevant when dealing with densely populated images with low signal‐to‐noise ratios—conditions that are often encountered in applications such as organelle tracking, virus particle tracking or single‐molecule imaging. We have developed a set of methods that can handle a wide variety of signal complexities. They are compiled into a free software package called Diatrack. Here we review its main features and utility in a range of applications, providing a survey of the dynamic imaging field together with recommendations for effective use. The performance of our framework is shown to compare favorably to a wide selection of custom‐developed algorithms, whether in terms of localization precision, processing speed or correctness of tracks.   相似文献   

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The use of radio frequency identification (RFID) technology is common in animal‐monitoring applications in the wild and in zoological and agricultural settings. RFID is used to track animals and to collect information about movements and other behaviors, as well as to automate or improve husbandry. Disney's Animal Kingdom® uses passive RFID technology to monitor nest usage by a breeding colony of northern carmine bee‐eaters. We implemented RFID technologies in various equipment configurations, initially deploying low‐frequency (LF) 125 kHz RFID and later changing to high‐frequency (HF) 13.56 MHz RFID technology, to monitor breeding behavior in the flock. We installed antennas connected to RFID readers at the entrances of nest tunnels to detect RFID transponders attached to leg bands as birds entered and exited tunnels. Both LF‐RFID and HF‐RFID systems allowed the characterization of nest visitation, including the timing of nest activity, breeding pair formation, identification of egg‐laying females, participation by nonresidents, and detection of nest disruptions. However, we collected a substantially larger volume of data using the increased bandwidth and polling speed inherent with HF‐RFID, which permitted tag capture of multiple birds simultaneously and resulted in fewer missed nest visits in comparison to LF‐RFID. Herein, we describe the evolution of the RFID setups used to monitor nest usage for more than 7 years, the types of data that can be gained using RFID at nests, and how we used these data to gain insights into carmine bee‐eater breeding behavior and improve husbandry.  相似文献   

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Understanding subcellular dynamic processes governing pathogenic mechanisms is a necessary step towards the development of new drugs and strategies against infectious diseases. Subcellular pathogenic mechanisms, such as viral invasion processes involve highly dynamic nanometric-scale objects and rapid molecular interactions that require the study of individual particle paths. Single-particle tracking methods allow visualizing and characterizing the dynamics of biological objects, and provide a straightforward and accurate means to understand subcellular processes. This review describes a number of particle-tracking methods in time-lapse microscopy sequences and provides examples of using such techniques to investigate mechanisms of host-pathogen interactions.  相似文献   

4.
Owing to the small size of mitochondria and the complexity of their motility patterns, mitochondrial tracking is technically challenging. Mitochondria are often tracked manually; however, this is time‐consuming and prone to measurement error. Here, we examined the suitability of four commercial and open‐source software alternatives for automated mitochondrial tracking in neurons compared with manual measurements. We show that all the automated tracking tools dramatically underestimated track length, mitochondrial displacement and movement duration, with reductions ranging from 45 to 77% of the values obtained manually. In contrast, mitochondrial velocity was generally overestimated. Only the number of motile mitochondria and their directionality were similar between strategies. Despite these discrepancies, we show that automated tools successfully detected transport alterations after applying an oxidant agent. Thus, automated methods appear to be suitable for assessing relative transport differences between experimental groups, but not for absolute quantification of mitochondrial dynamics. Although useful for objective and time‐efficient measurements of mitochondrial movements, results provided by automated methods should be interpreted with caution.   相似文献   

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In tracking analysis, the movement of cargos by motor proteins in axons is often represented by a time‐space plot termed a ‘kymograph’. Manual creation of kymographs is time‐consuming and complicated for cell biologists. Therefore, we developed KYMOMAKER, a simple system that automatically creates a kymograph from a movie without generating multiple time‐dissected movie stacks. In addition, KYMOMAKER can automatically extract faint vesicle traces, and can thereby effectively analyze cargos expressed at low levels in axons. A filter can be applied to remove traces of non‐physiological movements and to extract meaningful traces of anterograde or retrograde cargo transport. For example, only cargos that move at a speed of >0.4 µm/second for a distance of >1 µm can be included. Another function of KYMOMAKER is to create a color kymograph in which the color of the trace varies according to the position of the fluorescent particle in the axis perpendicular to the long axis of the axon. Such positional information is completely lost in conventional kymographs. KYMOMAKER is an open access program that can be easily used to analyze vesicle transport in axons by cell biologists who do not have specific knowledge of bioimage informatics .   相似文献   

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Seabirds spend most of their life at sea and have to possess a waterproof plumage to be able to sit on water for extended periods. We tracked juvenile Barau's petrels for the first time, when they leave their birth colony and found that half of the transmitters stopped soon after they first landed on the water off Réunion Island. We suspected from observation at sea that birds may have problems with the waterproofness of their plumage. Therefore during the next season we set up a simple protocol to assess waterproofness of the plumage of the birds just before they fledge. This protocol is based on the calculation of a wettability index expressed as the mass of water logged in the plumage after simulating the bird sitting on the sea surface. We found that at least one third of chicks ready to fledge gained more than 4 g of water in 20 s, indicating that plumage was not waterproof. Within a sample of birds having fledged and before reaching the sea surface, a similar proportion of birds had their plumage not waterproof. For the birds tracked, only those with an index indicating a waterproof plumage successfully dispersed after their first touch on the sea surface. We provide evidence of a possible major cause of mortality of juvenile seabirds that has not been described previously in wild seabirds, but could exist in other species. This issue may be a major cause of threat to the endangered Barau's petrel.  相似文献   

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This paper presents a method of monocular human motion tracking for estimation of hurdle clearance kinematic parameters. The analysis involved 10 image sequences of five hurdlers at various training levels. Recording of the sequences was carried out under simulated starting conditions of a 110 m hurdle race. The parameters were estimated using the particle swarm optimization algorithm and they are based on analysis of the images recorded with a 100 Hz camera. The proposed method does not involve using any special clothes, markers, inertial sensors, etc. As the quality criteria, the mean absolute error and mean relative error were used. The level of computed errors justifies the use of this method to estimate hurdle clearance parameters.  相似文献   

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