一种新的在线细胞显微观察仪及其在生化反应器上的应用研究

针对生化反应器应用条件,提出了用于生化反应器的在线细胞观察仪的基本技术要求。在比较了国内外现有的细胞在线显微工作原理后,研制了一种基于暗视场的新的显微细胞观察仪,介绍了其关键技术及结构。另外,原位在线显微细胞观察仪应用于酿酒酵母和哺乳动物细胞HEK293细胞的培养试验,在线细胞计数结果与离线细胞计数和细胞干重相比较,均有很好的相关性,表明此仪器基本满足使用要求。     

丝带凤蝶卵的显微观察

在丝带凤蝶生物学观察及人工饲养的实验研究中,采用体视显微镜对卵的外部形态特征进行显微摄影观察,了解丝带凤蝶不同季节产卵的不同特点及孵化前后颜色的变化,分析对其生活方式的影响,为丝带风蝶的饲养和分类等研究积累基础数据。     

烟草雄性不育的细胞形态学及生理生化研究进展(综述)

本文综述烟草雄性不育系及其同型保持系的花器官分化发育及形态特征、花药绒毡层特性、线粒体蛋白质差异及多种酶活性等细胞形态学和生理生化方面的研究进展。     

非常规介质中细胞生化反应研究进展

对完整细胞在非常规介质中的生物催化反应进行了回顾 ,分别总结了产物为醇 ,甾体 ,有机酸 ,生物大分子及其它各类反应的研究进展。并从溶剂和细胞两种角度对主要的研究方法进行了阐述。     

诊断桑蚕微粒子病的新仪器:显微直读仪

简介了显微直读仪的原理和结构,并介绍了应用显微直读仪代替普通显微镜检查桑蚕微粒子孢子的方法.     

新型多功能流式细胞仪的研制

一种新型流式细胞仪,利用一台装配有稳定的高压氙灯的落射荧光显微镜和光电倍增管以及计算机多道分析器组成的具有高分辨率和稳定性的流式细胞仪。其波长可在近紫外到近红外波段任意选择。既可用于流动样品,又可用于静止样品测定。用于流动样品测定时,喷嘴把经流体动力学方法会聚的样品流喷射到置于落射荧光显微镜物镜前方的显微镜盖玻片上,进行单细胞的快速定量分析。用于静止样品时,可用来精确测定细胞内各组分的含量。这种新型流式细胞仪可以在生物学研究和临床医学研究中得到广泛应用,有很好的发展前景。     

显微摄像条件对细胞形态测试结果的影响

目的:研究显微摄像条件对细胞形态测试结果的影响情况。方法:以苏木精-伊红(HE)染色的胃壁细胞为研究对象,分别在不同光亮度、对比度、饱和度和锐度的成像条件下进行显微摄像,采用图像分析软件(Image-Pro Plus 6.0)测试胃壁细胞的色度学和几何形态学参数,并对测试结果进行分析比较。结果:不同光亮度、对比度、饱和度组的胃壁细胞红、绿、蓝三基色差异均有统计学意义(P<0.05),其中红、绿、蓝基色值在高光亮度和高对比度组最高,同时红基色值在高饱和组最高,而绿、蓝基色值在高饱和组最低(P<0.05);不同锐度的胃壁细胞红、绿、蓝三基色差异没有统计学意义(P>0.05)。不同光亮度、对比度、饱和度和锐度的胃壁细胞的面积、周长、平均直径差异均没有统计学意义(P>0.05)。结论:光亮度、对比度和饱和度对细胞色度学参数影响明显,而对几何形态学参数无明显影响。     

玉米根冠脱落细胞中微丝分布的荧光显微观察（简报）

The detached root cap cells from maize seedlings comprised of round-shaped cells, ellipse-shaped cells and longitude-shaped cells. By using fluorescein-iso-thiocyanate-phalloidin(FITC-Ph) as fluorescent probe and treatment with cytochalasin B (CB) or HMC toxin, a host-specific toxin from Bipolaris (Helminthosporium) maydis race C, the distribution and variation of microfilaments (MFs) in detached cells were investigated. The results were as follows (i) the round-shaped cells had intense fluorescence, but the network of MFs was not distinct. There was clear MFs network in the cytoplasm of both ellipse-shaped and longitude-shaped cells. The distribution of MFs in detached cells seemed to be relevant to their shape and vigor. (ii) the fluorescence of detached cells of Charrua cytoplasmic male sterility(cms-C) maize was decreased after treatment with HMC-toxin. The cause of this outcome was unclear. We only obtained the pictures of the distorted protoplast membrane and dead cells owing to treatment with HMC-toxin. The distribution of MFs of detached cells of Normal(N) cytoplasmic maize was not affected by HMC-toxin, and their protoplasts shank slightly. (iii) CB could change the distribution of MFs in detached cells of both cms-C maize and N maize to disordered arrangement.     

孔雀鱼胚胎心血管发育的显微观察

心血管系统的组织分化研究是近年热点课题之一1.胚体透明且发育速度快的鱼类胚胎由于具备便于观察的特性2,正广泛用于脊椎动物造血系统的发生、血细胞生成、及血循环系统分化时序、调控因子等研究3-5.孔雀鱼(Poecilia reticulata Peters)是卵胎生(Ovoviviparity)鱼类的代表种,其卵子在雌鱼的泄殖腔内受精发育,长成鱼后再出生 ;同时孔雀鱼又是一种小型观赏鱼类,在实验室易于饲养,能不断提供实验用卵和胚胎。因此,在本实验室长期进行鱼类组织细胞培养的基础上6-11,制备孔雀鱼胚胎组织标本,并对早期胚胎及心血管系统进行了体外培养, 观察和比较了孔雀鱼胚胎心血管系统在体内、外的形成过程。       

一种便于丝状真菌显微观察的培养方法

在真菌培养过程中,对其个体形态与群体（菌落）形态进行实时观察与鉴定是很必要的。本文利用半培养基培养法结合显微操作技术,对丝状真菌个体与群体进行形态学观察。结果表明,该方法无需染色与制片,不破坏菌丝正常生长状态,可实时进行形态学检测,对多个菌种在自然生长状态下的菌丝与菌落特征进行观察,操作简单、方便、快捷,从而降低了成本及工作量。     

Beagle犬血清与胎牛血清生化特性及其对体外培养的人肺癌细胞生长的影响

目的比较Beagle犬血清与胎牛血清生化特性,探讨它们对体外培养的人肺癌细胞生长的影响。方法收集、制备Beagle犬血清与胎牛血清,采用全自动血液生化分析仪测定23项血清生化指标,比较分析两组血清生化特征差异。取对数期人肺癌细胞QG-56,分别培养于含10%FBS或Beagle-S的RPMI-1640合成培养基中,于第0、24、48、72、96、120和144 h各取出一块培养板进行检测,观察不同血清对细胞生长MTT曲线、细胞形态学的影响。结果FBS中CK、CK-MB、LDH、HBDH、GGT水平明显高于Beagle-S（P〈0.05）;CHE水平却明显低于Beagle-S,两者之间比较,均具有显著统计学意义。血清中的BUN、CRE、GLU、TP、ALB、ALT、ALP、TBIL、DBIL、TG、K^＋、Na^＋、Cl^-、Ca^2＋、Pi^3-水平在Beagle犬与胎牛间无显著性差异。MTT曲线和细胞形态学观察,两组于干预后24、48、72、96、120、144 h对QG-56细胞株生长的影响无显著性差异（P〈0.05）。结论Beagle犬的血清生化特性及其对体外肺癌细胞生长的影响与胎牛相似,可作为血清药理研究的优选动物之一。     

激光扫描共聚焦显微镜在植物学中的应用

激光扫描共聚焦显微镜(LSCM)是普通光学显微镜与激光和计算机及其相应的软件技术组合的产物,实现了连续光学切片,能在亚细胞水平观察细胞骨架的动态变化、细胞内特异蛋白、钙等离子的变化,并结合电生理等技术观察细胞生理活动与细胞形态及运动变化的相互关系。并广泛应用于生物三维结构重组及动态分析。本文综述了应用激光扫描共聚焦显微镜技术在植物细胞学、植物发育、组织化学以及基因表达、检测等领域取得的进展。     

似刺鳊鮈外周血细胞显微结构和血清生化指标的初步研究

似刺鳊(鮈)(Paracanthobrama guichenoti)外周血细胞可分为红血细胞、嗜中性粒细胞、单核细胞及大、小淋巴细胞和血栓细胞,未发现嗜酸性和嗜碱性粒细胞.外周血液中可观察到少量未成熟的及正在分裂的红血细胞.白细胞中,血栓细胞体积最小,嗜中性粒细胞体积最大;数量上,血栓细胞最多,而大淋巴细胞则最少.似刺鳊(鮈)血清生化指标测定结果显示,谷丙转氨酶(ALT)、胆固醇(CHOL)和总蛋白(TP)变化范围较大,且谷草转氨酶(AST)的水平较高,似刺鳊(鮈)雄鱼的血液生化指标相应值均显著低于雌鱼(P<0.05).     

色谱反应分离器中青霉素G的水解特性

6-氨基青霉烷酸(6-APA)是半合成青霉素的关键中间体,通常采用固定床反应器在青霉素酰化酶作用下水解青霉素来制备^[1]。由于该反应是典型的产物抑制可逆反应,工业生产中需不断加碱中和苯乙酸,以维持反应在最适pH条件下进行。为进一步提高青霉素的水解速率,除了维持最适反应条件外．消除产物抑制具有重要作用。近年来一些研究者提出采用伴有分离作用的反应分离组合系统水解青霉素 制取6-APA。Ishimura等^[2]采用电渗析膜耦合式生物反应器连续移除苯乙酸．将反应速率提高了近1倍;陈坚等^[3]研究了固定化酶-离子交换组合系统移除苯乙酸过程。本文报道青霉素在生化反应与液相谱分离过程相耦合而形成的色谱反应分离器中的水解特性。     

广西茶树资源生化成份多样性分析

对国家种质杭州茶树圃保存的来源于广西的98份茶树资源主要生化成分进行了评价鉴定和遗传多样性分析。结果发现,广西地方茶树资源的生化成分存在丰富的多样性和变异,平均遗传多样性指数达到1.90,平均变异系数达到25.8%。通过多变量的主成分分析,前7个主成分代表了茶树生化成分多样性的86.75%的信息。基于生化成分,把98份资源聚类划分为3个类群。第1类群大部分为红绿茶兼制的资源,第2类群大部分为适制红茶的资源,第3类群大部分为适制绿茶的资源。并从中筛选出一批生化成分特异的资源。     

Workflow for High-content,Individual Cell Quantification of Fluorescent Markers from Universal Microscope Data,Supported by Open Source Software

Advances in understanding the control mechanisms governing the behavior of cells in adherent mammalian tissue culture models are becoming increasingly dependent on modes of single-cell analysis. Methods which deliver composite data reflecting the mean values of biomarkers from cell populations risk losing subpopulation dynamics that reflect the heterogeneity of the studied biological system. In keeping with this, traditional approaches are being replaced by, or supported with, more sophisticated forms of cellular assay developed to allow assessment by high-content microscopy. These assays potentially generate large numbers of images of fluorescent biomarkers, which enabled by accompanying proprietary software packages, allows for multi-parametric measurements per cell. However, the relatively high capital costs and overspecialization of many of these devices have prevented their accessibility to many investigators.Described here is a universally applicable workflow for the quantification of multiple fluorescent marker intensities from specific subcellular regions of individual cells suitable for use with images from most fluorescent microscopes. Key to this workflow is the implementation of the freely available Cell Profiler software¹ to distinguish individual cells in these images, segment them into defined subcellular regions and deliver fluorescence marker intensity values specific to these regions. The extraction of individual cell intensity values from image data is the central purpose of this workflow and will be illustrated with the analysis of control data from a siRNA screen for G1 checkpoint regulators in adherent human cells. However, the workflow presented here can be applied to analysis of data from other means of cell perturbation (e.g., compound screens) and other forms of fluorescence based cellular markers and thus should be useful for a wide range of laboratories.     

Autohydrogenotrophic Denitrifying Microbial Community in a Glass Beads Biofilm Reactor

A glass bead biofilm reactor was operated using H₂ as an electron donor to remove nitrate at 150 mg NO₃–N l⁻¹ to below detection level. The microbial community in the glass beads biofilm reactor was investigated by using denaturing gradient gel electrophoresis (DGGE) and phylogenetic analysis. In DGGE analysis of the biofilm, five bands were dominant and indicated the presence of eight β-proteobacteria, one γ-proteobacteria and twelve clostridia. An unculturable Hydrogenophaga sp., which is a new genus of hydrogen-oxidizing bacterium was dominant in microbial community of the biofilm reactor.     

A Novel Gaussian Extrapolation Approach for 2D Gel Electrophoresis Saturated Protein Spots

Analysis of images obtained from two-dimensional gel electrophoresis (2D-GE) is a topic of utmost importance in bioinformatics research, since commercial and academic software available currently has proven to be neither completely effective nor fully automatic, often requiring manual revision and refinement of computer generated matches. In this work, we present an effective technique for the detection and the reconstruction of over-saturated protein spots. Firstly, the algorithm reveals overexposed areas, where spots may be truncated, and plateau regions caused by smeared and overlapping spots. Next, it reconstructs the correct distribution of pixel values in these overexposed areas and plateau regions, using a two-dimensional least-squares fitting based on a generalized Gaussian distribution. Pixel correction in saturated and smeared spots allows more accurate quantification, providing more reliable image analysis results. The method is validated for processing highly exposed 2D-GE images, comparing reconstructed spots with the corresponding non-saturated image, demonstrating that the algorithm enables correct spot quantification.