Live cell tracking based on cellular state recognition from microscopic images

The analysis of cell motion is an essential process in fundamental medical studies because most active cellular functions involve motion. In this paper, a computer-assisted motion analysis system is proposed for cell tracking. In the proposed tracking process, unlike in conventional tracking methods, cellular states referring to the cellular life cycle are defined and appropriate strategies are adopted for cells at different states. The use of cellular state recognition allows detection of possible cell division and hence can improve the robustness of cell tracking. Experimental results show that cells can be successfully segmented and tracked over a long period of time, and the proposed system is found to be as accurate as manual tracking. Various quantitative analyses and visualizations are used to represent cell motion, which demonstrates the usefulness of the proposed system in the study of cell dynamics.     

变胞机构的基因建模理论与构态进化分析

变胞机构的英文首创是“Metamorphic mechanisms”。这一概念借用了生物学中“Metamorphosis”,即变形、变拓扑态、可生长并进化演化。基于这一基本概念,并结合机构的基本杆组组成原理,首次提出变胞机构具有生物特性的组成原理,定义变胞元与变胞基因,并基于变胞基本单元建立变胞机构生物特性结构模型,分析基因进化引导的构型综合思想。通过分析变胞机构的构态转化特点,讨论变胞源机构及生成方法与过程,得出变胞机构构态演变过程生物学进化特性。深入研究变胞机构在变胞进化生长过程中构型要素的转化方式与蜕化方法,建立变胞机构学具有生物演变特征的机构组成原理及进化构态建模理论。在此基础上结合传统基本机构进化生成变胞机构构型给出分析示例,并基于工作阶段子机构之间进化为变胞机构过程中不同的自由度跃迁结果,对变胞源机构的进化计算分别进行讨论。     

Video-rate confocal reflection microscopy of neoplastic cells: Rate of intracellular movement and peripheral motility characteristic of neoplastic cell line (RSK4) with high degree of growth independence in vitro

Video-rate laser confocal interference reflection microscopy was used to demonstrate rapid motion of intracellular organelles and features at the cell periphery in a fully transformed neoplastic cell line, RSK4, and in four other neoplastic cell populations. In the RSK4 cells, vibrational and trafficking movements of intracellular particles at a rate greater than 25 Hz and ranging down to 5 Hz were recorded. Rapidly moving processes changed to ruffles, then microspikes, and previously undetectable ephemeral intercellular contacts were seen. Dynamic cyclical changes were revealed in the sizes of the podosomal close contacts of the transformed cells. The visibility of such features and the temporal and spatial resolution are improved over earlier methods. The fact that fast cellular and intracellular movements can be detected with this microscopic technique offers new possibilities in attempting to recognise differences between unimpaired living cells, and it may prove useful in the identification of malignant cells.     

大视场空间相机的像移速度场模型及卫星三轴姿态稳定度分析

针对大视场空间相机的像移补偿,建立了基于坐标变换和姿态动力学的离轴三反大视场空间相机通用像移速度场模型。建模过程中考虑了离轴三反光学系统的离轴角对像移模型的影响,推导了离轴三反大视场空间相机的像速场解析式。以某大视场空间相机为例,分析了3种典型成像姿态下焦面像移速度和偏流角的分布特点,研究了卫星姿态稳定度对相机成像质量的影响。分析表明,卫星三轴姿态稳定度的降低会导致相机焦面动态传递函数(MTF)下降,其中俯仰姿态稳定度对焦面动态MTF的影响最大;并且随着积分级数增加,下降会愈发明显。相机侧摆姿态成像时,对卫星姿态稳定度的要求更高。以传递函数下降5%为限,积分级数为96级的大视场空间相机,要求卫星姿态稳定度控制在0.001(°)/s以内。实验结果验证了文中对卫星姿态稳定度的分析,证明了像移速度场模型的准确性,为大视场空间相机像移补偿提供了可靠依据。     

Automatic detection and analysis of cell motility in phase‐contrast time‐lapse images using a combination of maximally stable extremal regions and Kalman filter approaches

Phase‐contrast illumination is simple and most commonly used microscopic method to observe nonstained living cells. Automatic cell segmentation and motion analysis provide tools to analyze single cell motility in large cell populations. However, the challenge is to find a sophisticated method that is sufficiently accurate to generate reliable results, robust to function under the wide range of illumination conditions encountered in phase‐contrast microscopy, and also computationally light for efficient analysis of large number of cells and image frames. To develop better automatic tools for analysis of low magnification phase‐contrast images in time‐lapse cell migration movies, we investigated the performance of cell segmentation method that is based on the intrinsic properties of maximally stable extremal regions (MSER). MSER was found to be reliable and effective in a wide range of experimental conditions. When compared to the commonly used segmentation approaches, MSER required negligible preoptimization steps thus dramatically reducing the computation time. To analyze cell migration characteristics in time‐lapse movies, the MSER‐based automatic cell detection was accompanied by a Kalman filter multiobject tracker that efficiently tracked individual cells even in confluent cell populations. This allowed quantitative cell motion analysis resulting in accurate measurements of the migration magnitude and direction of individual cells, as well as characteristics of collective migration of cell groups. Our results demonstrate that MSER accompanied by temporal data association is a powerful tool for accurate and reliable analysis of the dynamic behaviour of cells in phase‐contrast image sequences. These techniques tolerate varying and nonoptimal imaging conditions and due to their relatively light computational requirements they should help to resolve problems in computationally demanding and often time‐consuming large‐scale dynamical analysis of cultured cells.     

并联运动模拟系统的关键技术及研究现状

针对并联运动模拟系统的关键性技术进行了详细探讨,剖析了并联运动系统机构结构学理论、运动学理论、动力学理论以及控制策略的研究热点及难点,探讨了国内外对这些关键性技术的研究现状以及目前我国研制并联运动模拟系统所需研究的重点。本文的分析不仅提出了研制并联运动模拟系统的关键性技术方向,而且可以为并联运动模拟平台及其运动系统的进一步研究提供参考。     

旋转周期为曲柄二整周的平面双曲柄四杆机构

Grashof Neutral机构存在切换点(Change point)位置。通常认为机构在其切换点位置具有运动不确定性,其输入输出角速度比为0/0型不定式。根据运动连续性原理和洛毕达极限法则,首次导出了确定此类机构切换点(Change point)角速度比的解析公式。并通过机构运动分析、仿真,以及机构原型的制作和实验,证明:GrashofNeutral机构旋转通过切换点(Change point)时的运动是完全确定的,具其运动周期为驱动曲柄旋转二整周。特别是,当机构相对杆长差取很小值时,将输出有反向自锁特征和近似长停歇期的间歇运动,具有工业实用价值。     

3-PRC三维平移并联机器人机构的运动学分析

讨论了具有三平移自由度的3—PRC并联机器人机构的运动分析。基于机构运动约束方程,得到机构位置逆解的解析表达式;通过对刚体平面运动和点的合成运动分析,建立机构控制构件与动平台速度和加速度关系,并以显函数的形式表示;运动学解形式简单,便于实时控制。     

Development of a Leg Mechanism for Soft Landing Based on Biological Motion

With jumping mechanisms,soft landing motion is important to protect loads and the mechanisms.This study proposes a leg mechanism for soft landing based on biological motion.Human jumping motion with a load suggests a unique motion for soft landing.The landing model consists of two periods.Jerk is minimized in the first period and force is minimized in the second period.In comparison with other landing models,this model is specialized for soft landing motion protecting an objective part.Given all mechanisms ...     

相关系数分析在模糊图像参数识别中的应用

介绍一种在图像频域应用相关系数分析方法来识别模糊参数的方法。该方法用一系列同一类型但不同参数的模糊模型来第二次模糊图像,然后在频域中分析两次模糊图像之间的相关系数。实验结果表明,这种方法既适合于散焦模糊模型也适合于运动模糊模型的参数识别,有一定的准确性和计算上的简便性。     

Real-time quantitative elemental analysis and mapping: microchemical imaging in cell physiology

Recent advances in widely available microcomputers have made the acquisition and processing of digital quantitative X-ray maps of one to several cells readily feasible. Here we describe a system which uses a graphics-based microcomputer to acquire spectrally filtered X-ray elemental image maps that are fitted to standards, to display the image in real time, and to correct the post-acquisition image map with regard to specimen drift. Both high-resolution quantitative energy-dispersive X-ray images of freeze-dried cyrosections and low-dose quantitative bright-field images of frozen-hydrated sections can be acquired to obtain element and water content from the same intracellular regions. The software programs developed, together with the associated hardware, also allow static probe acquisition of data from selected cell regions with spectral processing and quantification performed on-line in real time. In addition, the unified design of the software program provides for off-line processing and analysing by several investigators at microcomputers remote from the microscope. The overall experimental strategy employs computer-aided imaging, combined with static probes, as an essential interactive tool of investigation for biological analysis. This type of microchemical microscopy facilitates studies in cell physiology and pathophysiology which focus on mechanisms of ionic (elemental) compartmentation, i.e. structure-function correlation at cellular and subcellular levels; it allows investigation of intracellular concentration gradients, of the heterogeneity of cell responses to stimuli, of certain fast physiological events in vivo at ultrastructural resolution, and of events occurring with low incidence or involving cell-to-cell interactions.     

平面3-RRR并联机器人运动分析的计算机模拟逼近法

提出一种新的平面3自由度并联机器人运动分析方法,计算机模拟逼近法。运用该方法进行了速度和加速度分析。先采用CAD的几何约束、尺寸约束、尺寸方程和尺寸驱动技术,构造平面3—RRR3自由度机器人的模拟机构。再用两个相同且彼此靠近的平面3自由度机器人模拟机构,构造速度模拟机构,求解动平台上确定点的速度和角速度。最后用两个相同且彼此靠近的平面3—RRR机器人速度模拟机构,构造加速度模拟机构,求解动平台上确定点的加速度和角加速度。计算机模拟结果与解析法的结果比较表明,该方法具有简单、快捷、直观、求解精度高的优点,而且可以完成复杂机构的运动分析,为多自由度机构运动分析提供了更有效的工具。     

Comparison of two automatic cell‐counting solutions for fluorescent microscopic images

Cell counting in microscopic images is one of the fundamental analysis tools in life sciences, but is usually tedious, time consuming and prone to human error. Several programs for automatic cell counting have been developed so far, but most of them demand additional training or data input from the user. Most of them do not allow the users to online monitor the counting results, either. Therefore, we designed two straightforward, simple‐to‐use cell‐counting programs that also allow users to correct the detection results. In this paper, we present the Cellcounter and Learn 123 programs for automatic and semiautomatic counting of objects in fluorescent microscopic images (cells or cell nuclei) with a user‐friendly interface. Although Cellcounter is based on predefined and fine‐tuned set of filters optimized on sets of chosen experiments, Learn 123 uses an evolutionary algorithm to determine the adapt filter parameters based on a learning set of images. Cellcounter also includes an extension for analysis of overlaying images. The efficiency of both programs was assessed on images of cells stained with different fluorescent dyes by comparing automatically obtained results with results that were manually annotated by an expert. With both programs, the correlation between automatic and manual counting was very high (R² < 0.9), although Cellcounter had some difficulties processing images with no cells or weakly stained cells, where sometimes the background noise was recognized as an object of interest. Nevertheless, the differences between manual and automatic counting were small compared to variations between experimental repeats. Both programs significantly reduced the time required to process the acquired images from hours to minutes. The programs enable consistent, robust, fast and accurate detection of fluorescent objects and can therefore be applied to a range of different applications in different fields of life sciences where fluorescent labelling is used for quantification of various phenomena. Moreover, Cellcounter overlay extension also enables fast analysis of related images that would otherwise require image merging for accurate analysis, whereas Learn 123's evolutionary algorithm can adapt counting parameters to specific sets of images of different experimental settings.     

软性磨粒流磨粒入射壁面过程及其加工特性研究

针对利用两相流模型无法计算高浓度固—液两相流固相颗粒撞击壁面时颗粒速度的问题,提出一种边界层内颗粒运动轨迹计算模型,基于Mixture两相流模型和Realizable k-ε湍流模型仿真计算结果,通过分析提取颗粒入射前速度、计算边界层厚度、建立边界层内速度场和颗粒运动分析可以得到颗粒撞击壁面时的速度和入射角度。分析加工表面动压力分布和磨粒体积分数分布,结合两种结构约束流道验证仿真结果与加工效果的对应关系。通过对试验结果的分析,为约束模块的设计提供依据。研究结果表明:磨粒入射速度、磨粒体积分数和加工表面所受动压力大小直接影响工件加工效果,并与材料去除量成正相关关系;在本次试验中选择的工艺参数导致加工材料去除量小,适合初始粗糙度低的工件表面加工,对于此次试验的初始粗糙度应在0.2μm以下;约束模块的设计除了要考虑磨粒流流场特性之外,还要对加工表面的原始加工痕迹作详细了解,为约束模块的设计及加工工艺参数提供参考。     

基于带漂移布朗运动的滚轮滑轨可靠性预测方法研究

针对滚轮滑轨的高耐磨性、少失效信息的特点,运用性能退化可靠性理论和随机过程方法,对滚轮滑轨的可靠性进行了研究。通过分析滚轮滑轨的磨损失效过程,建立了基于带漂移布朗运动的滚轮滑轨可靠度模型,并利用布朗运动特性研究了基于当前状态的寿命预测方法,从而实现滚轮滑轨可靠性的实时预测。实例证明了该方法的合理性和有效性。研究结果为滚轮滑轨的维修决策提供了理论方法。     

考虑混合阶次约束和驱动能力的伺服冲压运动设计

伺服冲压运动设计是伺服压力机实施伺服冲压作业的核心技术之一。提出NURBS运动曲线混合阶次插值方法,能任意给定冲压运动关键点上位置、速度或加速度中的一个或多个约束,以同时满足伺服冲压运动精确性和灵活性要求;提出快速生成给定时间的保压段冲压运动曲线算法;给出构造插值方程过程中合理设置切矢模长的取值方法,以避免运动曲线畸变;提出适当改变速度或加速度约束调节运动曲线性能的方法;在此基础上引入板料成形和伺服电动机驱动能力等约束,提出伺服冲压运动设计的一般流程,开发出交互式伺服冲压运动设计软件;以某型汽车翼子板拉深冲压运动设计为例进行了验证。     

Three‐dimensional cellular distribution in polymeric scaffolds for bone regeneration: a microCT analysis compared to SEM,CLSM and DNA content

In orthopaedic surgery the tissues damaged by injury or disease could be replaced using constructs based on biocompatible materials, cells and growth factors. Scaffold design, porosity and early colonization are key components for the implant success. From biological point of view, attention may be also given to the number, type and size of seeded cells, as well as the seeding technique and cell morphological and volumetric alterations. This paper describes the use of the microCT approach (to date used principally for mineralized matrix quantification) to observe construct colonization in terms of cell localization, and make a direct comparison of the microtomographic sections with scanning electron microscopy images and confocal laser scanning microscope analysis. Briefly, polycaprolactone scaffolds were seeded at different cell densities with MG63 osteoblastic‐like cells. Two different endpoints, 1 and 2 weeks, were selected for the three‐dimensional colonization and proliferation analysis of the cells. By observing all images obtained, in addition to a more extensive distribution of cells on scaffolds surfaces than in the deeper layers, cell volume increased at 2 weeks compared to 1 week after seeding. Combining the cell number quantification by deoxyribonucleic acid analysis and the single cell volume changes by confocal laser scanning microscope, we validated the microCT segmentation method by finding no statistical differences in the evaluation of the cell volume fraction of the scaffold. Furthermore, the morphological results of this study suggest that an effective scaffold colonization requires a precise balance between different factors, such as number, type and size of seeded cells in addition to scaffold porosity.     

航空摆扫相机转弯成像像移分析及补偿

为保证航空摆扫相机转弯成像过程中的成像质量,对其像移计算及补偿方法进行了研究。根据航空摆扫相机的成像原理,利用几何建模及速度矢量分解建立了转弯成像像移计算模型,给出了基于均值补偿的转弯前向像移补偿方法。转弯前向像移补偿分析表明:相机焦距为500 mm,曝光时间为0.01 s,速高比为0.02 rad/s,纵向视场角为10°,转弯角速度为0.5(°)/s时,最大前向像移补偿残差量为2.22μm;转弯角速度为1.5(°)/s时,最大前向像移补偿残差量为3.36μm。另外,转弯横向像移补偿分析表明:横向像移量随纵向视场角幅值的增加而增大,曝光时间为0.005 s,横向视场角为30(°),转弯角速度为1(°)/s时,横向像移量在纵向视场角为4.5°时达到3μm。转弯成像试飞实验结果表明:得到的图像像质优良,无几何形变,前向像移补偿良好,验证了本文提出的转弯成像像移补偿方法的正确性。     

An image processing system for cell behaviour studies in subconfluent cultures

DRIMAPS (digitally recorded interference microscopy with automatic phase-shifting) is a system that we have developed for quantifying the behaviour of cells in subconfluent cultures. The primary data generated by the system consist of phase-shifted interference (PSI) images which are accurate density maps of the distribution of dry mass (non-aqueous material) inside cells. Time-lapse sequences of PSI images may be viewed as movie sequences or processed in various ways to reveal many different aspects of the dynamics of cell growth and motile behaviour. Here we describe the image processing routines that are an integral part of the system and are required for four main functions: 1 initial calculation of the PSI images; 2 compensation of these images for instrumental distortion and instability; 3 identification and tracking of individual cells in a time-lapse sequence of PSI images; 4 extraction of cell behavioural data from a time-lapse sequence of PSI images. The first function converts standard interference microscope images into an image that accurately represents the optical phase-difference introduced by the specimen. The second function recalibrates a sequence of images by taking the cell-free region in each image as a reference plane of zero phase-difference. This is particularly necessary to compensate for the long-term instability of the Horn type of double-beam interference microscope, which has several advantages over other types of interference microscope for studying cell behaviour. The third function compares consecutive images in a sequence in order to trace the identity of individual cells throughout the sequence. Semi-automatic tracking, which allows close interaction with a human operator, is less prone to error than fully automatic tracking. The fourth function automatically extracts dynamic data from the identified cells. These data may include the true mass centroids of cells for translocation analysis and robust morphometric parameters for cell morphology examination. The integrated intensity of a cell is an accurate measure of cell mass and allows the growth (increase in dry mass) of individual cells to be studied. These data may be entered into a relational database of cell behaviour and a rule-based system allows efficient data access and analysis. Experiments with phase-contrast microscopy have revealed that many of these image processing methods are generally useful for cell behaviour studies using more conventional forms of microscopy.