电压钳控制电流钳的设计

研究证明,传统膜片钳放大器在电流钳模式下记录到的快速电压信号会存在失真,且造成失真的根本原因是由于膜片钳的探头电路设计.为了克服这些缺陷重新设计了一种探头,新探头电路不仅能像传统的电压跟随器一样测量瞬态电压,而且适用于传统的电压钳工作模式.此外,一种命名为电压钳控制的电流钳技术被应用来改进传统的膜片钳放大器.用可变的低通滤波器来调整电压钳模块的响应速度,从而在实现膜电位钳位的同时准确记录快速电压信号.桥平衡电路用来消除命令电流流过串联电阻时带来的电压误差.而传统膜片钳中的快电容补偿环节则被改进用来补偿电极分布电容和探头放大器输入电容并提高电流钳模式下系统的响应速度.细胞模型实验结果表明,改进后的膜片钳放大器能够满足电生理研究中快速电位变化测量的需要.     

用突起切断法分析MPP+对于多巴胺能神经细胞的损伤

对神经细胞在电压钳制方式下进行膜片钳记录受到胞体和突起空间钳位不一致的限制。本文尝试用突起切断方法区分胞体与突起的电压门控型钙电流,并据此对ＭＰＰ＋引起的钙电流变化进行分析。实验结果表明,突起上的电压门控钙通道的最大激活电压约为＋３０ｍＶ,而胞体的则为－１０～１０ｍＶ左右;ＭＰＰ＋损伤多巴胺能神经细胞后可导致电压门控型钙电流的抑制,这一抑制的Ｉ－Ｖ特性与突起钙电流Ｉ－Ｖ非常相近。由此提示,ＭＰＰ＋对于多巴胺能神经细胞的损伤更集中于其突起纤维。     

国产膜片钳仪研制成功

膜片钳(patch clamp)仪是从电压钳发展而来的新一代细胞电生理仪器。在直径约1μm的细胞膜片上,用膜片钳仪可以同时进行电压钳位和检测单个离子通道(ion channel)的电流(1 pA,10 kHz),这是迄今人们实现的最灵敏的仪器之一。1976年Neher & Sakmann首次实现单通道记录,Hamill等人(1981)对膜片钳的改进使之得以在世界推广,Sigworth(1983)和Rae & Levis(1984)对膜片钳仪的电子设计进行了深入的分析。从1988年起,我们根据Yale MK-V膜片钳仪电路图开始研制该仪器。     

利用膜片钳技术标记脑片神经元的方法

目的: 介绍一种利用膜片钳技术标记脑片神经元形态的方法。方法: 利用振动切片机切好实验目标部位的脑片,用含有Neurobiotin^TM Tracer的电极内液灌注玻璃微电极,并进行全细胞膜片钳记录;实验结束后将脑片先用4％多聚甲醛固定、漂洗,再用含有Streptavidin-Texas Red和Triton X-100的PB染色,2 h后即可用荧光显微镜观察着色的神经元。结果: 将细胞膜电压钳制在-70 mV,阶跃刺激后神经元表现为逐渐增大的膜电流。电流钳模式记录时,阶跃刺激使神经元去极化,达到阈电位后爆发动作电位。荧光显微镜下可看到胞体和主要突起清晰完整的神经元形态。结论: 本方法适用于在膜片钳实验后观察所记录的神经元的形态特征,操作方便,图像直观清晰。     

恒流灌注分离心肌细胞及膜片钳记录钾电流方法学探讨

目的探讨膜片钳实验中豚鼠、大鼠心肌细胞急性分离过程中的观察指标和影响因素,并进行膜片钳记录。方法使用改进的Langendorff恒流灌注法分离心肌细胞和全细胞膜片钳记录钾通道电流。结果分离即刻,存活细胞可达80%以上,复钙后约40%～50%呈静息状态,余出现自发性收缩,存活细胞能够进行膜片钳封接,于室温KB液中可成活8 h以上。结论使用恒流灌注法,观测灌注压的变化,可以较客观的掌握酶解消化的程度,利于获取生理状态良好的心肌细胞。且细胞能够进行膜片钳实验。     

小鼠腹腔巨噬细胞不完全失活的外向K~+电流的基本特性

采用膜片钳技术以全细胞方式在小鼠腹腔渗出巨噬细胞（ＰＥＭ）中记录到一种不完全失活的外向Ｋ＋电流（Ｉｏ）,该电流在膜电位正于－１０ｍＶ时激活,对Ｋ＋具有高度特异性,其半值电导电位Ｖ１／２为７９．５ｍＶ,在膜电位正于３０ｍＶ时,该电流失活,在６０－１２０ｍＶ的膜电位范围内,其失活时间常数τｉ与膜电位无关．随着胞外Ｋ＋离子浓度（［Ｋ＋］ｏ）升高,该电流的失活过程减慢。在生理电压范围内（－８０－０ｍＶ）,该电流缺乏稳态失活,且其失活不具有频率依赖性。胞外４－ＡＰ（３ｍｍｏｌ／Ｌ）、Ｂａ２＋（３ｍｍｏｌ／Ｌ）及ＴＥＡ（５ｍｍｏｌ／Ｌ）可抑制该电流,抑制率分别为８５％,６６％及３１％。胞外Ｚｎ２＋（１ｍｍｏｌ／Ｌ）可影响该电流活性,对该电流的抑制具有电压依赖性     

大鼠颈上神经节烟碱电流的整流及失敏

在培养的新生大鼠颈上神经节交感神经元标本上,用全细胞膜片钳技术研究了烟碱型乙酰胆碱受体（ｎＡＣｈＲ）通道的整流及失敏现象。烟碱激动剂引起的全细胞电流在膜电位为负值时随膜电位呈线性改变,而在膜电位达＋６０ｍＶ时仍不出现外向电流,表现出强烈的内向整流。ｎＡＣｈＲ通道电流存在失敏现象,即持续恒压喷射激动剂所引起的全细胞电流随时间呈指数衰减,不能保持在峰值水平,失敏随激剂浓度呈量效关系,膜电位的超极化也加     

脊神经损伤后钠电流的变化及其离子通道机制

目的：检测脊神经切断大鼠背根节（DRG）神经元重复放电能力和钠电流的变化,并研究介导其电流变化的钠通道亚型的表达情况。方法：脊神经切断术后2～8d慢性痛大鼠模型背根节急性分离,对中等直径DRG神经元运用全细胞膜片钳技术记录神经元放电和钠电流的变化。对背根节神经元进行RT-PCR检测,分析其钠通道亚型的表达情况。结果：电流钳下,实验组DRG神经元在电流刺激下产生重复放电,而对照组神经元多诱发单个动作电位,电压钳记录发现实验组背根节神经元快钠电流和持续性钠电流幅值均明显大于对照组,PCR结果显示,Nav1.3、Nav1.7和Nav1.8通道亚型mRNA表达显著增高。结论：钠通道介导了脊神经受损模型的DRG神经元兴奋性增高,持续性钠电流可能通过调节阈下膜电位振荡的产生调节神经元兴奋性。     

脊神经损伤后钠电流的变化及其离子通道机制

目的:检测脊神经切断大鼠背根节(DRG)神经元重复放电能力和钠电流的变化,并研究介导其电流变化的钠通道亚型的表达情况。方法:脊神经切断术后2～8d慢性痛大鼠模型背根节急性分离,对中等直径DRG神经元运用全细胞膜片钳技术记录神经元放电和钠电流的变化。对背根节神经元进行RT-PCR检测,分析其钠通道亚型的表达情况。结果:电流钳下,实验组DRG神经元在电流刺激下产生重复放电,而对照组神经元多诱发单个动作电位,电压钳记录发现实验组背根节神经元快钠电流和持续性钠电流幅值均明显大于对照组,PCR结果显示,Nav1.3、Nav1.7和Nav1.8通道亚型mRNA表达显著增高。结论:钠通道介导了脊神经受损模型的DRG神经元兴奋性增高,持续性钠电流可能通过调节阈下膜电位振荡的产生调节神经元兴奋性。     

Adaptive methods for growing electronic circuits on an imperfect synthetic matrix

Living systems can adapt to injuries and even heal themselves, an ability desirable also in synthetic systems. A method is presented for dynamically adapting the construction of an electronic circuit to hardware defects by formulating the process as a series of interactions between identical but specialized structures called supercells. The circuit components, including wires, can occupy any place in the hardware that has been determined to be free of defects. The circuit specification is reduced to a connected graph, with no positional information, and provided as a code repeated in each supercell. Using the code, supercells differentiate into circuit components in a late stage of the process, with highly adaptable physical location and organization; supercells also form the wires between circuit components. The structure and function of the system at three major levels is presented, the lowest cellular level, the supercell, and the target circuit level. Adaptation of circuit construction to defective hardware was observed for this method. Results obtained from this development process on simulated and real hardware with a variety of defect types and defect patterns are presented, as well as higher level simulations of the algorithm and its response to a wider range of defect patterns, amounts of hardware, and hardware to fault ratios.     

Circuit for the electromanipulation of plant protoplasts

An electric circuit for plant protoplast manipulation is described. The circuit used readily available materials and was designed for use in teaching. This integrated circuit can be placed in a single small box with controls for the aligning voltage, the aligning frequency, the pulse voltage, and the pulse timing. The circuit can be supplied by any suitable source of dc power and can be easily altered for individual requirements. The circuit, as presented here, can be assembled for less than $250.     

Equivalent Circuits as Related to Ionic Systems

The purpose of this paper is to clarify the relationship between certain “equivalent circuits” and the fundamental flux equations of Nernst and Planck. It is shown that as a direct algebraic consequence of these equations one may construct two types of equivalent circuits for a homogeneous (charged or uncharged) membrane. The one, which we term the “pure electrical equivalent circuit,” correctly predicts all of the electrical properties of the membrane for both steady and transient states. The other, which we call the “mixed equivalent circuit,” predicts the steady state I, Ψ characteristics of the membrane and the steady state ionic fluxes; it is not applicable to non-steady state properties or measurements. We emphasize that with regard to the portrayal of the physical basis of the properties of a homogeneous membrane, the mixed equivalent circuit can be misleading. This is particularly significant because this same circuit can also be used to depict a mosaic membrane, in which case the circuit gives a realistic pictorialization of the physical origin of the membrane properties. It is hoped that our analysis will be of aid to workers in electrophysiology who make use of equivalent circuit terminology in discussing the behavior of the plasma membrane.     

Clinical evaluation of an air embolism detection device

The clinical evaluation of the Sarns Air-Bubble Detector System in over 4,000 assorted operative procedures using cardiopulmonary bypass is described in this report. The system is designed to produce an alarm when a bolus of air enters the circuit. False alarms caused by electrical static within the operating room can be eliminated with a filter incorporated into the electrical circuit.     

Emergence of complex behaviour from simple circuit structures

The set of (feedback) circuits of a complex system is the machinery that allows the system to be aware of the levels of its crucial constituents. Circuits can be identified without ambiguity from the elements of the Jacobian matrix of the system. There are two types of circuits: positive if they comprise an even number of negative interactions, negative if this number is odd. The two types of circuits play deeply different roles: negative circuits are required for homeostasis, with or without oscillations, positive circuits are required for multistationarity, and hence, in biology, for differentiation and memory. In non-linear systems, a circuit can positive or negative (an 'ambiguous circuit', depending on the location in phase space. Full circuits are those circuits (or unions of disjoint circuits) that imply all the variables of the system. There is a tight relation between circuits and steady states. Each full circuit, if isolated, generates steady state(s) whose nature (eigenvalues) is determined by the structure of the circuit. Multistationarity requires the presence of at least two full circuits of opposite Eisenfeld signs, or else, an ambiguous circuit. We show how a significant part of the dynamical behaviour of a system can be predicted by a mere examination of its Jacobian matrix. We also show how extremely complex dynamics can be generated by such simple logical structures as a single (full and ambiguous) circuit.     

A structural and a functional aspect of stable information processing by the brain

Brain is an expert in producing the same output from a particular set of inputs, even from a very noisy environment. In this article a model of neural circuit in the brain has been proposed which is composed of cyclic sub-circuits. A big loop has been defined to be consisting of a feed forward path from the sensory neurons to the highest processing area of the brain and feed back paths from that region back up to close to the same sensory neurons. It has been mathematically shown how some smaller cycles can amplify signal. A big loop processes information by contrast and amplify principle. How a pair of presynaptic and postsynaptic neurons can be identified by an exact synchronization detection method has also been mentioned. It has been assumed that the spike train coming out of a firing neuron encodes all the information produced by it as output. It is possible to extract this information over a period of time by Fourier transforms. The Fourier coefficients arranged in a vector form will uniquely represent the neural spike train over a period of time. The information emanating out of all the neurons in a given neural circuit over a period of time can be represented by a collection of points in a multidimensional vector space. This cluster of points represents the functional or behavioral form of the neural circuit. It has been proposed that a particular cluster of vectors as the representation of a new behavior is chosen by the brain interactively with respect to the memory stored in that circuit and the amount of emotion involved. It has been proposed that in this situation a Coulomb force like expression governs the dynamics of functioning of the circuit and stability of the system is reached at the minimum of all the minima of a potential function derived from the force like expression. The calculations have been done with respect to a pseudometric defined in a multidimensional vector space.     

基于生理解剖知识的入睡机制神经元群网络模型研究

以生理解剖知识为基础,在已有的丘脑网状核细胞和丘脑皮质细胞间组成的入睡机制的两细胞环路模型［１］和由此两细胞环路组成的网络模型［２］的基础上,提出了增加皮层细胞在内的三种细胞组成的环路模型和网络模型,以使模型更符合近来认为睡眠机制是皮层和丘脑环路中出现特定的同步振荡的看法［３］。并能使模型的仿真结果可以和规定人体睡眠分期的脑电特征波相对应。这一网络模型的仿真结果,在一定条件下,确能在皮层细胞处出现符合睡眠分期中规定的标志入睡的纺锤波,这一初步结果,启示我们用模型仿真方法来进一步探讨睡眠机制和用模型仿真方法来进一步探讨人脑的微观神经元的电活动是如何通过同步振荡整合到宏观功能状态的某些信息处理过程的可能性。     

Functional Analysis of the Larval Feeding Circuit in Drosophila

The serotonergic feeding circuit in Drosophila melanogaster larvae can be used to investigate neuronal substrates of critical importance during the development of the circuit. Using the functional output of the circuit, feeding, changes in the neuronal architecture of the stomatogastric system can be visualized. Feeding behavior can be recorded by observing the rate of retraction of the mouth hooks, which receive innervation from the brain. Locomotor behavior is used as a physiological control for feeding, since larvae use their mouth hooks to traverse across an agar substrate. Changes in feeding behavior can be correlated with the axonal architecture of the neurites innervating the gut. Using immunohistochemistry it is possible to visualize and quantitate these changes. Improper handling of the larvae during behavior paradigms can alter data as they are very sensitive to manipulations. Proper imaging of the neurite architecture innervating the gut is critical for precise quantitation of number and size of varicosities as well as the extent of branch nodes. Analysis of most circuits allow only for visualization of neurite architecture or behavioral effects; however, this model allows one to correlate the functional output of the circuit with the impairments in neuronal architecture.     

Enabling flexibility on a dual head placement machine by optimizing platform-tray-feeder picking operations

The typical circuit card assembly line includes one or more dual head placement machines (DHPMs), which are capable of highly accurate placement and promise the flexibility to assemble a broad variety of circuit card types, each in minimal time. Each DHPM stages components for picking from feeder racks and platform tray feeders (PTFs), which are well suited for components that are large or odd-shaped, contributing to DHPM flexibility. The purpose of this paper is to present a model that can optimize operations that pick from a PTF and/or a feeder rack with the goal of enabling the flexibility needed to assemble a variety of circuit card types efficiently. The primary objective of this paper is a model to optimize operations that pick from a PTF and/or a feeder rack; a secondary objective is computational testing to evaluate the solvability of the model within reasonable run times.     

高频高压发生器逆变电路的控制方法

高频高压发生器是X线设备的核心部件之一。根据西班牙SEDECAL、美国QUANTUM、日本SHIMADZU与加拿大CPI四大公司生产的高频高压发生器的高压主回路逆变电路,结合高频高压发生器主电路的结构,分析了串联谐振逆变电路的各种控制方式及特点。它将为该类设备的维修、维护提供相应的技术指导。