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M. C. ADAMS A. MATOV D. YARAR S. L. GUPTON G. DANUSER & C. M. WATERMAN-STORER 《Journal of microscopy》2004,216(2):138-152
Fluorescent speckle microscopy (FSM) uses low levels of fluorescent proteins to create fluorescent speckles on cytoskeletal polymers in high‐resolution fluorescence images of living cells. The dynamics of speckles over time encode subunit turnover and motion of the cytoskeletal polymers. We sought to improve on current FSM technology by first expanding it to study the dynamics of a non‐polymeric macromolecular assembly, using focal adhesions as a test case, and second, to exploit for FSM the high contrast afforded by total internal reflection fluorescence microscopy (TIR‐FM). Here, we first demonstrate that low levels of expression of a green fluorescent protein (GFP) conjugate of the focal adhesion protein, vinculin, results in clusters of fluorescent vinculin speckles on the ventral cell surface, which by immunofluorescence labelling of total vinculin correspond to sparse labelling of dense focal adhesion structures. This demonstrates that the FSM principle can be applied to study focal adhesions. We then use both GFP‐vinculin expression and microinjected fluorescently labelled purified actin to compare quantitatively the speckle signal in FSM images of focal adhesions and the actin cytoskeleton in living cells by TIR‐FM and wide‐field epifluorescence microscopy. We use quantitative FSM image analysis software to define two new parameters for analysing FSM signal features that we can extract automatically: speckle modulation and speckle detectability. Our analysis shows that TIR‐FSM affords major improvements in these parameters compared with wide‐field epifluorescence FSM. Finally, we find that use of a crippled eukaryotic expression promoter for driving low‐level GFP‐fusion protein expression is a useful tool for FSM imaging. When used in time‐lapse mode, TIR‐FSM of actin and GFP‐conjugated focal adhesion proteins will allow quantification of molecular dynamics within interesting macromolecular assemblies at the ventral surface of living cells. 相似文献
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本文根据ITU-T最新的Q.2971协议,以及笔者所从事的实际课题,提出一整套B-ISDN第三层UNI信令的软件实现策略,其特点是能同时满足用户对点到点以及点到多点广播这两种通信方式的要求,而且可以动态配置。 相似文献
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介绍了最新的蓝牙技术规范4.0协议中低能耗部分的协议原理以及协议架构。讨论了在分层结构通信协议实现中遇到的主要问题和应对的策略。结合通信协议实现的通常策略,讨论了低能耗蓝牙4.0协议实现的方法与策略。 相似文献
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Miguel Martínez-Espronceda Jesús D. Trigo Santiago Led H. Gilberto Barrón-González Javier Redondo Alfonso Baquero Luis Serrano 《Computer methods and programs in biomedicine》2014
Experiences applying standards in personal health devices (PHDs) show an inherent trade-off between interoperability and costs (in terms of processing load and development time). Therefore, reducing hardware and software costs as well as time-to-market is crucial for standards adoption. The ISO/IEEE11073 PHD family of standards (also referred to as X73PHD) provides interoperable communication between PHDs and aggregators. Nevertheless, the responsibility of achieving inexpensive implementations of X73PHD in limited resource microcontrollers falls directly on the developer. Hence, the authors previously presented a methodology based on patterns to implement X73-compliant PHDs into devices with low-voltage low-power constraints. That version was based on multitasking, which required additional features and resources. This paper therefore presents an event-driven evolution of the patterns-based methodology for cost-effective development of standardized PHDs. The results of comparing between the two versions showed that the mean values of decrease in memory consumption and cycles of latency are 11.59% and 45.95%, respectively. In addition, several enhancements in terms of cost-effectiveness and development time can be derived from the new version of the methodology. Therefore, the new approach could help in producing cost-effective X73-compliant PHDs, which in turn could foster the adoption of standards. 相似文献
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HDLC协议一致性测试是保证不同厂家生产的HDLC通信设备之间实现互联、互通和互操作的前提;在对协议一致性测试基本工作原理进行简要分析的基础上,提出了HDLC协议一致性测试的基本方法以及测试过程中基于FSM模型的数据收发处理机制;并在实验室环境下构建了一致性测试硬件平台,设计了测试准备、测试执行和测试评估相关环节所涉及的基本软件功能模块,完成了HDLC协议一致性测试的基本功能;实际运行和测试结果表明,所设计的测试系统达到了预期结果;目前,该系统已成功应用于高速动车组上的通信测试,测试案例可以根据实际需求进行添加、删除和修改,提高了一致性测试的灵活性和完备性。 相似文献
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根据状态转换图输出属性为原态输出或次态输出,归纳和研究两类FSM电路设计方法.并从两个实例分析入手,明晰同步时序逻辑电路次态输出设计的可状态化简和寄存器输出特点,以及给出如何处理异步信号请求状态转换条件的通用方法.实际测试表明:系统具有稳定性好和结构简单等特点. 相似文献
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现有服务行为适配方法通常侧重于如何消除服务之间行为上的不相容而往往没有考虑用户的需求,因而需要对基于适配器的服务组合进行需求的可满足性验证,这使得基于适配器的服务组合过程复杂且低效.针对此问题,本文在适配器生成过程中考虑用户的需求,提出一种需求驱动的服务行为适配方法,并给出用于描述用户需求的形式模型的语法及语义.证明了利用本文方法所生成的适配器不仅可以消除服务之间行为上的不相容,并且保证基于适配器的服务组合的行为满足需求给出的时序约束,避免了对基于适配器的服务组合进行验证.通过一个贯穿全文的实例分析表明了方法的可行性和有效性 相似文献
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ContextFinite State Machine (FSM) inference from execution traces has received a lot of attention over the past few years. Various approaches have been explored, each holding different properties for the resulting models, but the lack of standard benchmarks limits the ability of comparing the proposed techniques. Evaluation is usually performed on a few case studies, which is useful for assessing the feasibility of the algorithm on particular cases, but fails to demonstrate effectiveness in a broad context. Consequently, understanding the strengths and weaknesses of inference techniques remains a challenging task.ObjectiveThis paper proposes CARE, a general, approach-independent, platform for the intensive evaluation of FSM inference techniques.MethodGrounded in a program specification scheme that provides a good control on the expected program structures, it allows the production of large benchmarks with well identified properties.ResultsThe CARE platform demonstrates the following features: (1) providing a benchmarking mechanism for FSM inference techniques, (2) allowing analysis of existing techniques w.r.t. a class of programs and/or behaviors, and (3) helping users in choosing the best suited approach for their objective. Moreover, our extensive experiments on different FSM inference techniques highlight that they do not behave in the same manner on every class of program. Characterizing different classes of programs thus helps understanding the strengths and weaknesses of the studied techniques.ConclusionExperiments reported in this paper show examples of use cases that demonstrate the ability of the platform to generate large and diverse sets of programs, which allows to carry out meaningful inference techniques analysis. The analysis strategies the CARE platform offers open new opportunities for program behavior learning, particularly in conjunction with model checking techniques. The CARE platform is available at http://care.lip6.fr. 相似文献
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领域外话语的开放性、口语化以及表达多样性,使得现有的限定领域口语对话系统不能很好地处理超出领域话语。该文提出了一种限定领域口语对话系统协处理方案,基于人工智能标记语言AIML,设计一套理解开放语义用户话语的理解模板,并对未匹配话语基于话语相似度进行理解模板分类,进而采用扩展有限状态自动机处理模式,结合对话流程上下文的状态及信息,实现理解模板到应答模板的转换,改变了单纯模板匹配方法在对话流程控制方面的相对缺失。中文手机导购领域的测试表明,该文所提出的协处理方法能有效地辅助口语对话系统完成限定领域完整对话流程,得到更好的用户满意度。
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