Search results for: Sahil Kapahi
18 Development of Restricted Formula SAE Intake Manifold Using 1D and Flow Simulations Based on Track Analysis
Authors: Sahil Kapahi
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A Formula SAE competition is characterized by typical track layouts having slaloms, tight corners and short straights, which favor a particular range of engine speed for a given set of gear ratios. Therefore, it is imperative that the power-train is optimized for the corresponding engine rpm band. This paper describes the process of designing, simulating and validating an air intake manifold for an inline four cylinder four-stroke internal combustion gasoline engine based on analysis of required vehicle performance. The requirements for the design of subject intake were set considering the rules of FSAE competitions and analysis of engine performance patterns for typical competition scenarios, carried out using OPTIMUMLAP software. Manifold geometry was optimized using results of air flow simulations performed on ANSYS CFX, and subsequent effect of this geometry on the engine was modeled using 1D simulation on Ricardo WAVE. A design was developed to meet the targeted performance standards in terms of engine torque output and volumetric efficiency. Finally, the intake manifold was manufactured and assembled onto the vehicle, and the engine output of the vehicle with the designed intake was studied using a dynamometer. The results of the dynamometer testing were then validated against predicted values derived from the Ricardo WAVE modeling and benefits to performance of the vehicle were established.Keywords: 1 D Simulation, air flow simulation, ANSYS CFX, four-stroke engine, OPTIMUM LAP, Ricardo WAVE
Procedia PDF Downloads 24617 Video Processing of a Football Game: Detecting Features of a Football Match for Automated Calculation of Statistics
Authors: Rishabh Beri, Sahil Shah
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We have applied a range of filters and processing in order to extract out the various features of the football game, like the field lines of a football field. Another important aspect was the detection of the players in the field and tagging them according to their teams distinguished by their jersey colours. This extracted information combined about the players and field helped us to create a virtual field that consists of the playing field and the players mapped to their locations in it.Keywords: Detect, Football, Players, Virtual
Procedia PDF Downloads 33016 Mapping Tunnelling Parameters for Global Optimization in Big Data via Dye Laser Simulation
Authors: Sahil Imtiyaz
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One of the biggest challenges has emerged from the ever-expanding, dynamic, and instantaneously changing space-Big Data; and to find a data point and inherit wisdom to this space is a hard task. In this paper, we reduce the space of big data in Hamiltonian formalism that is in concordance with Ising Model. For this formulation, we simulate the system using dye laser in FORTRAN and analyse the dynamics of the data point in energy well of rhodium atom. After mapping the photon intensity and pulse width with energy and potential we concluded that as we increase the energy there is also increase in probability of tunnelling up to some point and then it starts decreasing and then shows a randomizing behaviour. It is due to decoherence with the environment and hence there is a loss of ‘quantumness’. This interprets the efficiency parameter and the extent of quantum evolution. The results are strongly encouraging in favour of the use of ‘Topological Property’ as a source of information instead of the qubit.Keywords: big data, optimization, quantum evolution, hamiltonian, dye laser, fermionic computations
Procedia PDF Downloads 19315 An Exploratory Research on Childhood Sexual Victimization and Its Psychological Impacts
Authors: Urwah Ali
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The aim of this study is to carry out a meta-analysis in order to establish an overall international figure and to summarize the evidence relating to the possible relationship between child sexual abuse and subsequent mental and physical health outcomes. A systematic review was conducted using the HEC Digital Library, Pub Med, PsycINFO and SAHIL databases published after 2010 containing empirical data pertaining to CSA. Out of 124 articles assessed for eligibility, 32 studies provided evidence of a relationship between sexual child maltreatment and various health outcomes for use in subsequent meta-analyses. Statistical significance associations were observed between childhood sexual victimization and psychological problems in their adulthood [odds ratio (OR) = 1.5; 95%Cl 3.07–4.43]. For most studies included for meta-analysis, the odds ratio falls above 1.00, indicating that patients having history of childhood sexual victimization were more likely to develop psychological disorders.Keywords: abuse, sexual abuse, childhood sexual abuse, mental health
Procedia PDF Downloads 40714 Designing Expressive Behaviors to Improve Human-Robot Relationships
Authors: Sahil Anand, John Luetke, Nikhil Venkatesh, Dorothy Wong
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Trust plays an important role in building and sustaining long-term relationships between people. In this paper, we present a robot that communicates using nonverbal behaviors such as facial expressions and body movements. Our study reports on an experiment in which participants were asked to team up with the robot to perform specific tasks. We varied the expressivity of the robot and measured the effects on trust, quality of interactions as well as on the praising and punishing behavior of the participant towards the robot. We found that participants developed a stronger affinity towards the expressive robot, but did not show any significant differences in the level of trust. When the same robot made mistakes, participants unconsciously punished it with lesser intensity compared to the neutral robot. The results emphasize the role of expressive behaviors on participant’s perception of the robot and also on the quality of interactions between humans and robots.Keywords: human-robot interaction, nonverbal communication, relationships, social robot, trust
Procedia PDF Downloads 37013 Enhancing the Work of Art through Fashion Attire
Authors: A. N. Roslen, S. A. Syed-Sahil, A. Musavir
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In Malaysia, there are only few fashion designers who are inspired by the work of artists when creating their collections. The researchers confirmed this statement by interviewing fashion experts in Malaysia. The objectives of this study are to: 1. Investigate the acceptance of fashion inspired by the work of art among consumers. 2. Encourage more designers to use work of art as their inspirations. 3. Promote Malaysian Artists through fashion. Thus, the researchers interviewed Malaysian fashion designers, image consultants, and one famous Malaysian Artist (Awang Damit). All of them had agreed that the fashion inspired by the work of art in Malaysia has a long way to go. Therefore, the researchers’ aim is to attract more fashion designers to use the work of local artists in their creations. The researchers had used interview, survey and experimentation as methods of this study. In the experimentation procedure, paintings of local artist, Awang Damit was used as a source of inspiration in creating a design Line. The result of this study had shown that fashion inspired by work of art is acknowledged and accepted by the designers and consumers.Keywords: art, fashion, inspiration, local artist
Procedia PDF Downloads 45912 Fundamental Research Dissension between Hot and Cold Chamber High Pressure Die Casting
Authors: Sahil Kumar, Surinder Pal, Rahul Kapoor
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This paper is focused on to define the basic difference between hot and cold chamber high pressure die casting process which is not fully defined in a research before paper which we have studied. The pressure die casting is basically defined into two types (1) Hot chamber Die Casting (2) Cold chamber Die Casting. Cold chamber die casting is used for casting alloys that require high pressure and have a high melting temperature, such as brass, aluminum, magnesium, copper based alloys and other high melting point nonferrous alloys. Hot chamber die casting is suitable for casting zinc, tin, lead, and low melting point alloys. In hot chamber die casting machine, the molten metal is an integral pan of the machine. It mainly consists of hot chamber and gooseneck type metal container made of cast iron. This machine is mainly used for low melting alloys and alloys of metals like zinc, lead etc. Metals and alloys having a high melting point and those which are having an affinity for iron cannot be cast by this machine, which could otherwise attack the shot sleeve and damage the machine.Keywords: hot chamber die casting, cold chamber die casting, metals and alloys, casting technology
Procedia PDF Downloads 61611 Comparison of User Experience in VR When Hand Gestures Are Used vs. Using Controller
Authors: Sanu Muhammed C., Nihal Vadakkan, Sahil Athrij, Sasi Gopalan
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Currently, the content entertainment industry is dominated by mobile devices. As the trends slowly shift towards AR/VR applications, the user experience on these devices should be considered, and we are trying to improve user experience in VR. This paper proposes a survey-based solution to improve user experience in VR. By creating a VR environment where users can move a ball from one position to another using a remote controller and another VR environment where users can move a ball from one place to another using hand gestures/ By allowing a set of audience to use these two environments, we can get their feedback. There are two steps in this comparison, 1) Using Hand Gestures To Move Ball In VR Environment: Here, we create a VR environment where two baskets are there, and one ball will be there in a basket. Here users can transfer the ball to another basket using hand gestures. They will be able to move the ball using hand gestures. 2) Using Remote Control To Move Ball In VR Environment: Here, we create a VR environment where two baskets are there, and one ball will be there in a basket. Here users can transfer the ball to another basket using a remote control. They will be able to move the ball using a remote controller. The above two environments are given to users to experience, and their responses will be recorded to compare the user experience in the above two environments.Keywords: virtual reality, user experience, hand gestures, remote control
Procedia PDF Downloads 14410 The Effect of the Addition of Additives on the Properties of Bisamide Organogels
Authors: Elmira Ghanbari, Jan Van Esch, Stephen J. Picken, Sahil Aggarwal
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Organogels are formed by the assembly of low molecular weight gelators (LMWG) into fibrous structures. The assembly of these molecules into crystalline fibrous structures occurs as a result of reversible interactions such as π-stacking, hydrogen-bonding, and van der Waals interactions. Bisamide organogelators with two amide groups have been used as one of LMWGs which show efficient assembly behavior via hydrogen bonding for network formation, the formation of a crystalline network for solvent entrapment. In this study, different bisamide gelators with different lengths of alkyl chains have been added to the bisamide parent gels. The effect of the addition of bisamide additives on the gelation of bisamide gels is described. Investigation of the thermal properties of the gels by differential scanning calorimetry and dropping ball techniques indicated that the bisamide gels can be formed by the addition of a high concentration of the second bisamide components. The microstructure of the gels with different gelator components has been visualized with scanning electron microscopy (SEM) which has shown systematic woven, platelet-like, and a combination of those morphologies for different gels. Examining the addition of a range of bisamide additives with different structural characteristics than the parent bisamide gels has confirmed the effect of the molecular structure on the morphology of the bisamide gels and their final properties.Keywords: bisamide organogelator additives, gel morphology, gel properties, self-assembly
Procedia PDF Downloads 2009 Gaits Stability Analysis for a Pneumatic Quadruped Robot Using Reinforcement Learning
Authors: Soofiyan Atar, Adil Shaikh, Sahil Rajpurkar, Pragnesh Bhalala, Aniket Desai, Irfan Siddavatam
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Deep reinforcement learning (deep RL) algorithms leverage the symbolic power of complex controllers by automating it by mapping sensory inputs to low-level actions. Deep RL eliminates the complex robot dynamics with minimal engineering. Deep RL provides high-risk involvement by directly implementing it in real-world scenarios and also high sensitivity towards hyperparameters. Tuning of hyperparameters on a pneumatic quadruped robot becomes very expensive through trial-and-error learning. This paper presents an automated learning control for a pneumatic quadruped robot using sample efficient deep Q learning, enabling minimal tuning and very few trials to learn the neural network. Long training hours may degrade the pneumatic cylinder due to jerk actions originated through stochastic weights. We applied this method to the pneumatic quadruped robot, which resulted in a hopping gait. In our process, we eliminated the use of a simulator and acquired a stable gait. This approach evolves so that the resultant gait matures more sturdy towards any stochastic changes in the environment. We further show that our algorithm performed very well as compared to programmed gait using robot dynamics.Keywords: model-based reinforcement learning, gait stability, supervised learning, pneumatic quadruped
Procedia PDF Downloads 3158 A Framework for Improving Trade Contractors’ Productivity Tracking Methods
Authors: Sophia Hayes, Kenny L. Liang, Sahil Sharma, Austin Shema, Mahmoud Bader, Mohamed Elbarkouky
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Despite being one of the most significant economic contributors of the country, Canada’s construction industry is lagging behind other sectors when it comes to labor productivity improvements. The construction industry is very collaborative as a general contractor, will hire trade contractors to perform most of a project’s work; meaning low productivity from one contractor can have a domino effect on the shared success of a project. To address this issue and encourage trade contractors to improve their productivity tracking methods, an investigative study was done on the productivity views and tracking methods of various trade contractors. Additionally, an in-depth review was done on four standard tracking methods used in the construction industry: cost codes, benchmarking, the job productivity measurement (JPM) standard, and WorkFace Planning (WFP). The four tracking methods were used as a baseline in comparing the trade contractors’ responses, determining gaps within their current tracking methods, and for making improvement recommendations. 15 interviews were conducted with different trades to analyze how contractors value productivity. The results of these analyses indicated that there seem to be gaps within the construction industry when it comes to an understanding of the purpose and value in productivity tracking. The trade contractors also shared their current productivity tracking systems; which were then compared to the four standard tracking methods used in the construction industry. Gaps were identified in their various tracking methods and using a framework; recommendations were made based on the type of trade on how to improve how they track productivity.Keywords: labor productivity, productivity tracking methods, trade contractors, construction
Procedia PDF Downloads 1917 Utilizing Laser Cutting Method in Men's' Custom-Made Casualwear
Authors: M A. Habit, S. A. Syed-Sahil, A. Bahari
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Abstract—Laser cutting is a method of manufacturing process that uses laser in order to cut materials. It provides and ensures extreme accuracy which has a clean cut effect, CO2 laser dominate this application due to their good- quality beam combined with high output power. It comes with a small scale and it has a limitation in cutting sizes of materials, therefore it is more appropriate for custom- made products. The same laser cutting machine is also capable in cutting fine material such as fine silk, cotton, leather, polyester, etc. Lack of explorations and knowledge besides being unaware about this technology had caused many of the designers not to use this laser cutting method in their collections. The objectives of this study are: 1) To identify the potential of laser cutting technique in Custom-Made Garments for men’s casual wear: 2) To experiment the laser cutting technique in custom made garments: 3) To offer guidelines and formula for men’s custom- made casualwear designs with aesthetic value. In order to achieve the objectives, this research has been conducted by using mixed methods which are interviews with two (2) local experts in the apparel manufacturing industries and interviews via telephone with five (5) local respondents who are local emerging fashion designers, the questionnaires were distributed to one hundred (100) respondents around Klang Valley, in order to gain the information about their understanding and awareness regarding laser cutting technology. The experiment was conducted by using natural and man- made fibers. As a conclusion, all of the objectives had been achieved in producing custom-made men’s casualwear and with the production of these attires it will help to educate and enhance the innovation in fine technology. Therefore, there will be a good linkage and collaboration between the design experts and the manufacturing companies.Keywords: custom-made, fashion, laser cut, men’s wear
Procedia PDF Downloads 4416 Hybrid Velocity Control Approach for Tethered Aerial Vehicle
Authors: Lovesh Goyal, Pushkar Dave, Prajyot Jadhav, GonnaYaswanth, Sakshi Giri, Sahil Dharme, Rushika Joshi, Rishabh Verma, Shital Chiddarwar
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With the rising need for human-robot interaction, researchers have proposed and tested multiple models with varying degrees of success. A few of these models performed on aerial platforms are commonly known as Tethered Aerial Systems. These aerial vehicles may be powered continuously by a tether cable, which addresses the predicament of the short battery life of quadcopters. This system finds applications to minimize humanitarian efforts for industrial, medical, agricultural, and service uses. However, a significant challenge in employing such systems is that it necessities attaining smooth and secure robot-human interaction while ensuring that the forces from the tether remain within the standard comfortable range for the humans. To tackle this problem, a hybrid control method that could switch between two control techniques: constant control input and the steady-state solution, is implemented. The constant control approach is implemented when a person is far from the target location, and error is thought to be eventually constant. The controller switches to the steady-state approach when the person reaches within a specific range of the goal position. Both strategies take into account human velocity feedback. This hybrid technique enhances the outcomes by assisting the person to reach the desired location while decreasing the human's unwanted disturbance throughout the process, thereby keeping the interaction between the robot and the subject smooth.Keywords: unmanned aerial vehicle, tethered system, physical human-robot interaction, hybrid control
Procedia PDF Downloads 975 Storm-water Management for Greenfield Area Using Low Impact Development Concept for Town Planning Scheme Mechanism
Authors: Sahil Patel
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Increasing urbanization leads to a concrete forest. The effects of new development practices occur in the natural hydrologic cycle. Here the concerns have been raised about the groundwater recharge in sufficient quantity. With further development, porous surfaces reduce rapidly. A city like Ahmedabad, with a non-perennial river, is 100% dependent on groundwater. The Ahmedabad city receives its domestic use water from the Narmada river, located about 200 km away. The expenses to bring water is much higher. Ahmedabad city receives annually 800 mm rainfall, and mostly this water increases the local level waterlogging problems; after that, water goes to the Sabarmati river and merges into the sea. The existing developed area of Ahmedabad city is very dense, and does not offer many chances to change the built form and increase porous surfaces to absorb storm-water. Therefore, there is a need to plan upcoming areas with more effective solutions to manage storm-water. This paper is focusing on the management of stormwater for new development by retaining natural hydrology. The Low Impact Development (LID) concept is used to manage storm-water efficiently. Ahmedabad city has a tool called the “Town Planning Scheme,” which helps the local body drive new development by land pooling mechanism. This paper gives a detailed analysis of the selected area (proposed Town Planning Scheme area by the local authority) in Ahmedabad. Here the development control regulations for individual developers and some physical elements for public places are presented to manage storm-water. There is a different solution for the Town Planning scheme than that of the conventional way. A local authority can use it for any area, but it can be site-specific. In the end, there are benefits to locals with some financial analysis and comparisons.Keywords: water management, green field development, low impact development, town planning scheme
Procedia PDF Downloads 1294 Wet Processing of Algae for Protein and Carbohydrate Recovery as Co-Product of Algal Oil
Authors: Sahil Kumar, Rajaram Ghadge, Ramesh Bhujade
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Historically, lipid extraction from dried algal biomass remained a focus area of the algal research. It has been realized over the past few years that the lipid-centric approach and conversion technologies that require dry algal biomass have several challenges. Algal culture in cultivation systems contains more than 99% water, with algal concentrations of just a few hundred milligrams per liter ( < 0.05 wt%), which makes harvesting and drying energy intensive. Drying the algal biomass followed by extraction also entails the loss of water and nutrients. In view of these challenges, focus has shifted toward developing processes that will enable oil production from wet algal biomass without drying. Hydrothermal liquefaction (HTL), an emerging technology, is a thermo-chemical conversion process that converts wet biomass to oil and gas using water as a solvent at high temperature and high pressure. HTL processes wet algal slurry containing more than 80% water and significantly reduces the adverse cost impact owing to drying the algal biomass. HTL, being inherently feedstock agnostic, i.e., can convert carbohydrates and proteins also to fuels and recovers water and nutrients. It is most effective with low-lipid (10--30%) algal biomass, and bio-crude yield is two to four times higher than the lipid content in the feedstock. In the early 2010s, research remained focused on increasing the oil yield by optimizing the process conditions of HTL. However, various techno-economic studies showed that simply converting algal biomass to only oil does not make economic sense, particularly in view of low crude oil prices. Making the best use of every component of algae is a key for economic viability of algal to oil process. On investigation of HTL reactions at the molecular level, it has been observed that sequential HTL has the potential to recover value-added products along with biocrude and improve the overall economics of the process. This potential of sequential HTL makes it a most promising technology for converting wet waste to wealth. In this presentation, we will share our experience on the techno-economic and engineering aspects of sequential HTL for conversion of algal biomass to algal bio-oil and co-products.Keywords: algae, biomass, lipid, protein
Procedia PDF Downloads 2133 The Effect of the Structural Arrangement of Binary Bisamide Organogelators on their Self-Assembly Behavior
Authors: Elmira Ghanbari, Jan Van Esch, Stephen J. Picken, Sahil Aggarwal
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Low-molecular-weight organogelators form gels by self-assembly into the crystalline network which immobilizes the organic solvent. For single bisamide organogelator systems, the effect of the molecular structure on the molecular interaction and their self-assembly behavior has been explored. The spatial arrangement of bisamide molecules in the gel-state is driven by a combination of hydrogen bonding and Van der Waals interactions. The hydrogen-bonding pattern between the amide groups of bisamide molecules is regulated by the number of methylene spacers; the even number of methylene spacers between two amide groups, in even-spaced bisamides, leads to the antiparallel position of amide groups within a molecule. An even-spaced bisamide molecule with antiparallel amide groups can make two pairs of hydrogen bonding with the molecules on the same plane. The odd-spaced bisamide with a parallel directionality of amide groups can form four independent hydrogen bonds with four other bisamide molecules on different planes. The arrangement of bisamide molecules in the crystalline state and the interaction of these molecules depends on the molecular structure, particularly the parity of the spacer length between the amide groups in the bisamide molecule. In this study, the directionality of amide groups has been exploited as a structural characteristic to affect the arrangement of molecules in the crystalline state and produce different binary bisamide gelators with different degrees of crystallinities. Single odd- and even-spaced single bisamides were synthesized and blended to produce binary bisamide organogelators to be characterized in order to understand the effect of the different directionality of amide groups on the molecular interaction in the crystalline state. The pattern of molecular interactions between these blended molecules, mixing or phase separation, has been monitored via differential scanning calorimetry (DSC) and crystallography techniques; X-ray powder diffraction (XRD) and Small-angle X-ray scattering (SAXS). The formation of lamellar structures for odd- and even-spaced bisamide gelators was confirmed by using SAXS and XRD techniques. DSC results have shown that binary bisamide organogelators with different parity of methylene spacers (odd-even binary blends) have a higher tendency for phase separation compared to the binary bisamides with the same parity (odd-odd or even-even binary blends). Phase separation in binary odd-even bisamides was confirmed by the presence of individual (100) reflections of odd and even lamellar structures. The structural characteristic of bisamide organogelators, the parity of spacer length in binary systems, is a promising tool to control the arrangement of molecules and their crystalline structure.Keywords: binary bisamide organogelators, crystalline structure, phase separation, self-assembly behavior
Procedia PDF Downloads 1852 Protocol for Dynamic Load Distributed Low Latency Web-Based Augmented Reality and Virtual Reality
Authors: Rohit T. P., Sahil Athrij, Sasi Gopalan
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Currently, the content entertainment industry is dominated by mobile devices. As the trends slowly shift towards Augmented/Virtual Reality applications the computational demands on these devices are increasing exponentially and we are already reaching the limits of hardware optimizations. This paper proposes a software solution to this problem. By leveraging the capabilities of cloud computing we can offload the work from mobile devices to dedicated rendering servers that are way more powerful. But this introduces the problem of latency. This paper introduces a protocol that can achieve high-performance low latency Augmented/Virtual Reality experience. There are two parts to the protocol, 1) In-flight compression The main cause of latency in the system is the time required to transmit the camera frame from client to server. The round trip time is directly proportional to the amount of data transmitted. This can therefore be reduced by compressing the frames before sending. Using some standard compression algorithms like JPEG can result in minor size reduction only. Since the images to be compressed are consecutive camera frames there won't be a lot of changes between two consecutive images. So inter-frame compression is preferred. Inter-frame compression can be implemented efficiently using WebGL but the implementation of WebGL limits the precision of floating point numbers to 16bit in most devices. This can introduce noise to the image due to rounding errors, which will add up eventually. This can be solved using an improved interframe compression algorithm. The algorithm detects changes between frames and reuses unchanged pixels from the previous frame. This eliminates the need for floating point subtraction thereby cutting down on noise. The change detection is also improved drastically by taking the weighted average difference of pixels instead of the absolute difference. The kernel weights for this comparison can be fine-tuned to match the type of image to be compressed. 2) Dynamic Load distribution Conventional cloud computing architectures work by offloading as much work as possible to the servers, but this approach can cause a hit on bandwidth and server costs. The most optimal solution is obtained when the device utilizes 100% of its resources and the rest is done by the server. The protocol balances the load between the server and the client by doing a fraction of the computing on the device depending on the power of the device and network conditions. The protocol will be responsible for dynamically partitioning the tasks. Special flags will be used to communicate the workload fraction between the client and the server and will be updated in a constant interval of time ( or frames ). The whole of the protocol is designed so that it can be client agnostic. Flags are available to the client for resetting the frame, indicating latency, switching mode, etc. The server can react to client-side changes on the fly and adapt accordingly by switching to different pipelines. The server is designed to effectively spread the load and thereby scale horizontally. This is achieved by isolating client connections into different processes.Keywords: 2D kernelling, augmented reality, cloud computing, dynamic load distribution, immersive experience, mobile computing, motion tracking, protocols, real-time systems, web-based augmented reality application
Procedia PDF Downloads 721 Optimized Electron Diffraction Detection and Data Acquisition in Diffraction Tomography: A Complete Solution by Gatan
Authors: Saleh Gorji, Sahil Gulati, Ana Pakzad
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Continuous electron diffraction tomography, also known as microcrystal electron diffraction (MicroED) or three-dimensional electron diffraction (3DED), is a powerful technique, which in combination with cryo-electron microscopy (cryo-ED), can provide atomic-scale 3D information about the crystal structure and composition of different classes of crystalline materials such as proteins, peptides, and small molecules. Unlike the well-established X-ray crystallography method, 3DED does not require large single crystals and can collect accurate electron diffraction data from crystals as small as 50 – 100 nm. This is a critical advantage as growing larger crystals, as required by X-ray crystallography methods, is often very difficult, time-consuming, and expensive. In most cases, specimens studied via 3DED method are electron beam sensitive, which means there is a limitation on the maximum amount of electron dose one can use to collect the required data for a high-resolution structure determination. Therefore, collecting data using a conventional scintillator-based fiber coupled camera brings additional challenges. This is because of the inherent noise introduced during the electron-to-photon conversion in the scintillator and transfer of light via the fibers to the sensor, which results in a poor signal-to-noise ratio and requires a relatively higher and commonly specimen-damaging electron dose rates, especially for protein crystals. As in other cryo-EM techniques, damage to the specimen can be mitigated if a direct detection camera is used which provides a high signal-to-noise ratio at low electron doses. In this work, we have used two classes of such detectors from Gatan, namely the K3® camera (a monolithic active pixel sensor) and Stela™ (that utilizes DECTRIS hybrid-pixel technology), to address this problem. The K3 is an electron counting detector optimized for low-dose applications (like structural biology cryo-EM), and Stela is also a counting electron detector but optimized for diffraction applications with high speed and high dynamic range. Lastly, data collection workflows, including crystal screening, microscope optics setup (for imaging and diffraction), stage height adjustment at each crystal position, and tomogram acquisition, can be one of the other challenges of the 3DED technique. Traditionally this has been all done manually or in a partly automated fashion using open-source software and scripting, requiring long hours on the microscope (extra cost) and extensive user interaction with the system. We have recently introduced Latitude® D in DigitalMicrograph® software, which is compatible with all pre- and post-energy-filter Gatan cameras and enables 3DED data acquisition in an automated and optimized fashion. Higher quality 3DED data enables structure determination with higher confidence, while automated workflows allow these to be completed considerably faster than before. Using multiple examples, this work will demonstrate how to direct detection electron counting cameras enhance 3DED results (3 to better than 1 Angstrom) for protein and small molecule structure determination. We will also show how Latitude D software facilitates collecting such data in an integrated and fully automated user interface.Keywords: continuous electron diffraction tomography, direct detection, diffraction, Latitude D, Digitalmicrograph, proteins, small molecules
Procedia PDF Downloads 106