Cell analysis system based on immunomagnetic cell selection and alignment followed by immunofluorescent analysis using compact disk technologies

BACKGROUND: Although the flow cytometer has become the standard in cell analysis, it has limitations. Recently, we introduced a new cell analysis method based on immunomagnetic selection and aligning of cells. No flow system is needed and cell analysis can be performed in whole blood. METHODS: Whole blood is incubated with fluorescent labels and immunomagnetic nanoparticles. The blood is injected into a capillary that is in a strong magnetic field. The immunomagnetic-labeled cells move upward and align themselves along ferromagnetic lines present on the upper surface of the capillary. An optical focus and tracking system analogous to that used in a conventional compact disk player focuses a 635-nm laser-diode on the magnetically aligned cells. The emitted fluorescence signals are projected on two photomultipliers. Allophycocyanin (APC)-labeled CD4 (CD4-APC) and Cyanin5.5 (Cy5.5)-labeled CD8 (CD8-Cy5.5) antibodies and Oxazine750, all red excited, are used as fluorescent labels. RESULTS: A differential white blood cell count performed in whole blood is obtained using the CD4-APC in combination with Oxazine750. The results are compared with the Technicon-H1 hematology analyzer. Correlation coefficients of 0.91 for neutrophilic granulocytes, 0.93 for lymphocytes, 0.93 for monocytes, and 0.96 for eosinophilic granulocytes were obtained. Immunofluorescence is demonstrated using CD4-APC and CD8-Cy5.5. The absolute counts obtained for CD4+ and CD8+ are compared with the Coulter Epics XL flow cytometer. Correlation coefficients of, respectively, 0.91 and 0.94 were obtained. CONCLUSION: We conclude that our system is as capable as a standard flow cytometer or hematology analyzer for a reliable routine white blood cell analysis, including immunophenotyping, and can be used as an easy-to-handle disposable white blood cell test.     

Microfluidic analysis of antibody specificity in a compact disk format

A new and flexible technology for high throughput analysis of antibody specificity and affinity is presented. The method is based on microfluidics and takes advantage of compact disks (CDs) in which the centrifugal force moves fluids through microstructures containing immobilized metal affinity chromatography columns. Analyses are performed as a sandwich assay, where antigen is captured to the column via a genetically attached His6-tag. The antibodies to be analyzed are applied onto the columns. Thereafter, fluorescently labeled secondary antibodies recognize the bound primary antibodies, and detection is carried out by laser-induced fluorescence. The CDs contain 104 microstructures enabling analysis of antibodies against more than 100 different proteins using a single CD. Importantly, through the three-dimensional visualization of the binding patterns in a column it is possible to separate high affinity from low affinity binding. The method presented here is shown to be very sensitive, flexible and reproducible.     

Cell deposition system based on laser guidance

We have designed a laser cell deposition system that employs the phenomenon of laser guidance to place single cells at specific points in a variety of in vitro environments. Here, we describe the components of the system: the laser optics, the deposition chamber, the microinjection cell feeding system and our custom system control software application. We discuss the requirements and challenges involved in laser guidance of cells and how our present system overcomes these challenges. We demonstrate that the patterning system is accurate within one micrometer by repeatedly depositing polymer microspheres and measuring their position. We demonstrate its ability to create highly defined living patterns of cells by creating a defined pattern of neurons with neurite extensions displaying normal function. We found that the positional accuracy of our system is smaller than the variations in cell size and pattern disruptions that occur from normal cell movement during substrate adhesion. The laser cell deposition system is a potentially useful tool that can be used to achieve site- and time-specific placement of an individual cell in a cell culture for the systematic investigation of cell-cell and cell-extracellular matrix interactions.     

Workflow management system based on WEB technology

Through the analysis of workflow management system based on Web, the work, following WFMC workflow model, proposed a universal workflow management system with the combination of specific application requirements of enterprises. Workflow management system based on Web was described from system function, software structure, system structure, workflow engine and security. The actual case of the design of aeronautical structure showed that workflow system performance can be greatly improved by using Web technology.     

基于文献计量的微藻收获技术研究分析

微藻是能以自养模式固定二氧化碳,生成生物能源的原料,对可持续发展具有重要意义。微藻也能以异养模式生长,用于废水处理和积累高附加值物质。目前,微藻收获的成本占总成本的20%~30%。微藻收获技术已经成为研究热点。本文从文献计量的角度分析了各国微藻收获的研究进展以及我国的研究现状,并展望了微藻收获技术发展趋势,为进一步研究提供参考。     

A new cancer diagnostic system based on a CDK profiling technology

A series of molecular pathological investigations of the molecules that stimulate the cyclin dependent kinases (CDK1, 2, 4, and 6) have led to enormous accumulation of knowledge of the clinical significance of these molecules for cancer diagnosis. However, the molecules have yet to be applied to clinical cancer diagnosis, as there is no available technology for application of the knowledge in a clinical setting. We hypothesized that the direct measurement of CDK activities and expressions (CDK profiling) might produce clinically relevant values for the diagnosis. This study investigated the clinical relevance of CDK profiling in gastrointestinal carcinoma tissues by using originally developed expression and activity analysis methods. We have established novel methods and an apparatus for analyzing the expression and activities of the CDK molecules in lysate of tumor tissue in a clinical setting, and examined 30 surgically dissected gastrointestinal carcinomas and corresponding normal mucosal specimens. We demonstrate here that remarkably elevated CDK2 activity is evident in more than 70% of carcinoma tissues. Moreover, a G1-CDK activity profiling accurately mirrored the differences in proliferation between tumor and normal colonic tissues. Our results suggest that CDK profiling is a potent molecular-clinical approach to complement the conventional pathological diagnosis, and to further assist in the individualized medications.     

An analytical system based on a compact flow cytometer for DNA fragment sizing and single-molecule detection.

BACKGROUND: Previous reports have demonstrated accurate DNA fragment sizing of linear DNA fragments, from 564 to approximately 4 x 10(5) bp, in a flow system. B-phycoerythrin (B-PE), commonly used in conventional cytometric applications that require high-sensitivity, was the first fluorophore detected in flow at the single-molecule level. METHODS: Dilute solutions of stained DNA fragments or B-PE were analyzed in a simplified, compact flow system, with enhanced performance and lower cost, utilizing a solid-state laser and a single-photon sensing avalanche photodiode detector (SSAPD). Extensive data processing and display software, developed specifically for the photon-counting data stream, extracts correlated height, width, and area features from bursts of photons due to discrete molecules passing through the sensing region in the flow channel. RESULTS: DNA fragment sizing in flow has now been demonstrated for SYTOX-orange-stained fragments ranging in size over 3.4 orders of magnitude, from 125 to 5 x 10(5) bp. For Lambda bacteriophage DNA (lambda DNA; 48.5 kbp) a CV of 1.2 % has been achieved. Analysis of a femtomolar B-PE solution demonstrates that the bursts of photons from individual molecules can be baseline-resolved with 0.5 mW of laser power at a signal to noise ratio (SNR) of approximately 30, with approximately 100 photons detected from each molecule. CONCLUSIONS: A compact, low-power, high-sensitivity system detects DNA fragments as small as 125 bp or individual B-PE molecules in a flowing liquid stream. Demonstrated linearity, sensitivity, and resolution indicate that <1.0 mW of laser power is optimal, permitting further miniaturization of the system and additional cost reduction. Comprehensive analytical software exploits the standard cytometric paradigm of multiple 2D graphs and gating to extract features from classes of individually analyzed biomolecules. This complete system is thus poised to engage high-sensitivity applications not amenable to conventional flow cytometric instrumentation.     

A brain-inspired compact cognitive mapping system

In many simultaneous localization and mapping (SLAM) systems, the map of the environment grows over time as the robot explores the environment. The ever-growing map prevents long-term mapping, especially in large-scale environments. In this paper, we develop a compact cognitive mapping approach inspired by neurobiological experiments. Mimicking the firing activities of neighborhood cells, neighborhood fields determined by movement information, i.e. translation and rotation, are modeled to describe one of the distinct segments of the explored environment. The vertices with low neighborhood field activities are avoided to be added into the cognitive map. The optimization of the cognitive map is formulated as a robust non-linear least squares problem constrained by the transitions between vertices, and is numerically solved efficiently. According to the cognitive decision-making of place familiarity, loop closure edges are clustered depending on time intervals, and then batch global optimization of the cognitive map is performed to satisfy the combined constraint of the whole cluster. After the loop closure process, scene integration is performed, in which revisited vertices are removed subsequently to further reduce the size of the cognitive map. The compact cognitive mapping approach is tested on a monocular visual SLAM system in a naturalistic maze for a biomimetic animated robot. Our results demonstrate that the proposed method largely restricts the growth of the size of the cognitive map over time, and meanwhile, the compact cognitive map correctly represents the overall layout of the environment. The compact cognitive mapping method is well suitable for the representation of large-scale environments to achieve long-term robot navigation. Electronic supplementary materialThe online version of this article (10.1007/s11571-020-09621-6) contains supplementary material, which is available to authorized users.     

Transcriptome analysis of Verticillium lecanii based on RNA-Seq technology

[目的]蜡蚧轮枝菌是一种防治植物病原物和害虫的生防菌,弄清其致病的分子基础具有重要的意义。[方法]利用Illumina HiSeq技术测序蜡蚧轮枝菌的cDNA文库,并进行RNA-Seq序列拼接和功能注释。[结果]共获得1634670条高品质reads,碱基GC含量48.50%。14856条Unigene经组装拼接后,在Nr、Swiss-Prot、COG和KEGG数据库中分别比对得到1467、9379、6518和4188条Unigenes。采用GO分类工具将21403条Unigenes分成24个功能组,主要包含在细胞组成（1369）、分子功能（1679）和生物学过程（1928）3个大类里,在COG数据库注释的基础上,把6518条Unigenes分成38个功能组,通过GO和COG丰度识别分类,聚类的主要是一般功能预测、次生代谢产物的合成、运输和分解代谢以及信号转导机制起作用的独立基因。能与KEGG路径匹配的108条Unigenes中大部分和新陈代谢途径及次生代谢产物的合成有关。[结论]这些结果将有助于找寻蜡蚧轮枝菌的致病因子,从而丰富其致病机理。     

基于SLAF-seq技术的苗药八爪金龙遗传分析

为探究苗药八爪金龙群体遗传进化和亲缘关系的远近,该研究利用简化基因组测序技术(SLAFseq)对42份八爪金龙样品进行测序,获得多态性SLAF标签。同时采用GATK和SAMtools软件在多态性SLAF中检测单核苷酸多态性(SNP)分子标记,并利用SNP分子标记分析八爪金龙样品间的遗传分化关系。结果表明:(1) 42份八爪金龙共获得246.35 Mb reads,测序质量值Q30的平均值为95.66%,GC含量的平均值为41.14%。(2)通过生物信息学的分析,获得1 769 265个SLAF标签,其中379 829个多态性SLAF标签,共开发2 299 640个SNPs分子标记。(3)利用开发的SNPs数据构建八爪金龙的系统发育树,42份八爪金龙分成两个大的类群。第一类群为细柄百两和原变种百两金;第二类群由贵州朱砂根、红凉伞、湖北朱砂根和江西朱砂根组成。江西朱砂根与其余群体关系较远,有明显的分群现象。该研究从基因组水平揭示不同地区八爪金龙资源之间的遗传关系,为八爪金龙种质资源的鉴定和遗传多样性分析提供了理论基础,所开发的SNP位点可进一步用于挖掘与品质、抗逆性等相关的基因。     

Cell lineage analysis of the Drosophila peripheral nervous system

The peripheral nervous system (PNS) of Drosophila provides a very well-characterized model system for studying the genes involved in basic processes of neurogenesis. Because of its simplicity and stereotyped pattern, each cell of the PNS can be individually identified and the phenotypic consequences of mutations can be studied in detail. Thus, some of the genetic mechanisms leading to the formation of type I sensory organs, the external, bristle-type sensory organs (es), and the internal, stretch-receptive chordotonal organs (ch) have been elucidated. Each sensory organ seems to be generated by a stereotyped pattern of cell division of individual ectodermal precursor cells. Recent advances in cell lineage analysis of the PNS have provided a detailed picture of almost all the lineages in the PNS, including those giving rise to the type II sensory neurons, also known as multiple dendritic (md) neurons. This knowledge will be instrumental in the precise characterization of the phenotypes associated with mutations in known and new genes and their interactions which determine cell fate decisions during neurogenesis. Here, we describe and compare three recently developed methods by which cell lineages have been assessed: single cell transplantation, bromodeoxyuridine (BrdU) incorporation studies, and the flp/FRT recombinase system from yeast. In the light of a more complete knowledge of the PNS lineages, we will discuss the effects of known mutations that alter neuronal cell fates. © 1996 Wiley-Liss, Inc.     

A compact fiber‐optic probe‐based singlet oxygen luminescence detection system

This paper presents a novel compact fiberoptic based singlet oxygen near‐infrared luminescence probe coupled to an InGaAs/InP single photon avalanche diode (SPAD) detector. Patterned time gating of the single‐photon detector is used to limit unwanted dark counts and eliminate the strong photosensitizer luminescence background. Singlet oxygen luminescence detection at 1270 nm is confirmed through spectral filtering and lifetime fitting for Rose Bengal in water, and Photofrin in methanol as model photosensitizers. The overall performance, measured by the signal‐to‐noise ratio, improves by a factor of 50 over a previous system that used a fiberoptic‐coupled superconducting nanowire single‐photon detector. The effect of adding light scattering to the photosensitizer is also examined as a first step towards applications in tissue in vivo.

     

基于空间技术的烤烟种植生态环境综合评价分析

光照、气温、土壤类型、降水量、土壤含水量、海拔、坡度、重金属污染等自然生态环境对烤烟的分布、生长发育及品质有重要影响, 同时也是烤烟种植优化配置必不可少的基础信息。基于空间技术的烤烟种植生态环境综合评价分析的目的就是应用遥感、地理信息等空间技术, 实现烤烟种植区域生态环境信息的三维立体采集及烤烟种植生态环境适宜性空间分布信息评价。主要研究内容是分析影响烤烟种植及品质的生态因子, 应用ARCGIS空间分析功能获得海拔、坡度因子, 应用常规的遥感模型反演植被指数、地表温度、土壤含水量和太阳辐射因子, 应用Kriging插值加密地球化学数据建立烤烟种植生态环境综合评价指标体系。通过基于空间技术的烤烟种植生态环境适宜性等级空间分布优化分析结果与实验区烤烟种植示范基地和烤烟种植区域的调整结果进行对比, 可知该方法是有效的。     

基于能值的污水处理系统环境影响分析

污水处理厂及其“产物”(处理水和脱水污泥)的处理处置系统共同构成了污水处理综合系统,它从整体上涵盖了污水处理的全过程.污水处理综合系统在构建和运行的过程中主要包括两方面的环境影响:1)物质资源的消耗;2)排放污染物对环境的危害,而后者在既往有关的污水处理系统分析中往往被忽略.为了更全面地了解污水处理过程的环境影响状况,本文以能值分析理论为基础对污水处理综合系统进行分析,同时,结合生态足迹理论对系统的可持续性进行分析.结果表明:水体污染物的环境影响能值远大于大气污染物,且主要受氨氮排放量的影响;污水处理综合系统的能值消耗主要为废物流能值和本地可更新资源能值;“处理厂系统+填埋系统”的能值利用效率最高,“处理厂系统+中水回用系统+焚烧系统”的能值利用效率最低;“处理厂系统+中水回用系统+填埋系统”的环境可持续性最好,而“处理厂系统+焚烧系统”的可持续性最差.     

基于能值分析的广东省生态经济系统综合研究

运用能值理论与分析方法,从自然子系统、社会子系统、经济子系统3个侧面,系统与子系统2个层面,对广东省生态经济系统1990年以来的发展态势进行了定量分析.结果表明,广东省属于高度发展地区,系统高度依赖于反馈能值的投入,虽然污染治理卓有成效,但并未从整体上解决环境压力不断增大的矛盾,区域自身可持续发展能力低.广东省对于国际市场的信赖性日益提高,但在大量资源型产品出口和高技术产品进口的不等价交换中处于实际的亏损地位.依靠科技,提高产出品附加值,发挥落后地区自然和人力资源优势,以及发达地区的经济拉动力,是全面提高区域可持续发展能力的关键.     

基于3S技术的广州市生态安全景观格局分析

生态安全已经成为国家安全的组成部分,成为区域、国家可持续发展研究的重要内容.基于生态安全的数字评价图,从生态安全的等级配置和空间结构的角度,分别运用景观指数和半变异函数的方法探讨广州市生态安全的景观格局与动态特征.景观指数法分析结果显示,广州市生态安全现状以一般安全为主,生态安全曾在1995年受创,但其总的趋势良好;从景观破碎化、形态复杂性以及景观多样性来看,生态安全的时间与空间分异明显.变异函数的结果表明,生态安全的可预测性高,研究区生态安全可能存在等级结构.     

A high-temporal resolution technology for dynamic proteomic analysis based on 35S labeling

As more and more research efforts have been attracted to dynamic or differential proteomics, a method with high temporal resolution and high throughput is required. In present study, a (35)S in vivo Labeling Analysis for Dynamic Proteomics (SiLAD) was designed and tested by analyzing the dynamic proteome changes in the highly synchronized A549 cells, as well as in the rat liver 2/3 partial hepatectomy surgery. The results validated that SiLAD technique, in combination with 2-Dimensional Electrophoresis, provided a highly sensitivity method to illustrate the non-disturbed endogenous proteins dynamic changes with a good temporal resolution and high signal/noise ratio. A significant number of differential proteins can be discovered or re-categorized by this technique. Another unique feature of SiLAD is its capability of quantifying the rate of protein expression, which reflects the cellular physiological turn points more effectively. Finally, the prescribed SiLAD proteome snapshot pattern could be potentially used as an exclusive symbol for characterizing each stage in well regulated biological processes.     

基于图像识别技术的金针菇表型高通量采集与分析

金针菇Flammulina filiformis作为国内产销量大、工厂化生产程度高的食用菌种类之一,其育种工作因表型信息采集效率低下而受到限制.本研究以金针菇子实体可见光图像为信息来源,基于育种工作对表型信息的需求,利用图像识别技术和深度学习模型,提出了金针菇高通量表型信息采集分析方法,并开发了相应分析系统软件.利用该...