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1.
动物胚胎的玻璃化冻存是一项新的深低温冷冻方法,已成功地冻存了多种动物和人的胚胎,具有较高的实际应用和研究价值。本文结合本实验室的研究工作,论述玻璃化冻存方法在动物胚胎保存中的应用,不同保护剂和玻璃化溶液对胚胎存活率的影响,分析影响玻璃化冻存的主要因素包括冷冻保护剂与玻璃化溶液的选择、预平衡过程、冷却和复温速率控制、去玻璃化的避免、冻存后玻璃化溶液的洗脱等。  相似文献   

2.
动物组织细胞的玻璃化冻存是一项新的深低温冷冻保存方法,已成功地冻存了多种动物细胞,具有较高的实际应用和研究价值。本文论述几种主要组织细胞玻璃化冻存方法的研究及其进展,如小鼠胰岛和卵母细胞,大鼠卵巢、牛胚泡、牛卵母细胞、猪胚胎、人胚胎等,不同冷冻保护剂,玻璃化溶液和冷冻程序对细胞存活率的影响。并提出了玻璃化冻存动物以及组织细胞的问题与展望。  相似文献   

3.
梁艺勇 《大众科技》2012,(1):221-222
高峰矿深部由于种种原因,目前只能采用小盲斜井串车提升矿石,靠人工装卸车。而在人工装卸车的过程中,一旦发生飞斗事故,轻则造成设备损伤,重则造成人员伤亡。因此,在高峰矿开展小盲斜井防跑车装置研究,设计适合高峰矿深部小盲斜井防跑车装置,对于保证高峰矿深部开采安全、提高综合开采效益具有重要意义。文章重点研究变坡点下20米的挡车器。  相似文献   

4.
A variety of methods have been used to introduce chemicals into a stream or to mix two or more streams of different compositions using microfluidic devices. In the following paper, the introduction of cryoprotective agents (CPAs) used during cryopreservation of cells in order to protect them from freezing injuries and increase viability post thaw is described. Dimethylsulphoxide (DMSO) is the most commonly used CPA. We aim to optimize the operating conditions of a two-stream microfluidic device to introduce a 10% vol/vol solution of DMSO into a cell suspension. Transport behavior of DMSO between two streams in the device has been experimentally characterized for a spectrum of flow conditions (0.7 < Re < 10), varying initial donor stream concentrations, (1% vol/vol < Co < 15% vol/vol) and different flow rate fractions (0.23 < fq < 0.77). The outlet cell stream concentration is analyzed for two different flow configurations: one with the cell stream flowing on top of the DMSO-rich donor stream, and the other with the cell stream flowing beneath the heavy DMSO-laden stream. We establish a transition from a diffusive mode of mass transfer to gravity-influenced convective currents for Atwood numbers (At) in the range of (1.7 × 10−3 < At < 3.1 × 10−3) for the latter configuration. Flow visualization with cells further our understanding of the effect of At on the nature of mass transport. Cell motion studies performed with Jurkat cells confirm a high cell recovery from the device while underscoring the need to collect both the streams at the outlet of the device and suggesting flow conditions that will help us achieve the target DMSO outlet concentration for clinical scale flow rates of the cell suspension.  相似文献   

5.
In this work, we conduct a computational study on the loading of cryoprotective agents into cells in preparation for cryopreservation. The advantages of microfluidics in cryopreserving cells include control of fluid flow parameters for reliable cryoprotectant loading and reproducible streamlined processing of samples. A 0.25 m long, three inlet T-junction microchannel serves as an idealized environment for this process. The flow field and concentration distribution are determined from a computational fluid dynamics study and cells are tracked as inert particles in a Lagrangian frame. These particles are not confined to streamlines but can migrate laterally due to the Segre-Sildeberg effect for particles in a shear flow. During this tracking, the local concentration field surrounding the cell is monitored. This data are used as input into the Kedem-Katchalsky equations to numerically study passive solute transport across the cell membrane. As a result of the laminar flow, each cell has a unique pathline in the flow field resulting in different residence times and a unique external concentration field along its path. However, in most previous studies, the effect of a spatially varying concentration field on the transport across the cell membrane is ignored. The dynamics of this process are investigated for a population of cells released from the inlet. Using dimensional analysis, we find a governing parameter α, which is the ratio of the time scale for membrane transport to the average residence time in the channel. For α <  = 0.224, cryoprotectant loading is completed to within 5% of the target concentration for all of the cells. However, for α > 0.224, we find the population of cells does not achieve complete loading and there is a distribution of intracellular cryoprotective agent concentration amongst the population. Further increasing α beyond a value of 2 leads to negligible cryoprotectant loading. These simulations on populations of cells may lead to improved microfluidic cryopreservation protocols where more consistent cryoprotective agent loading and freezing can be achieved, thus increasing cell survival.  相似文献   

6.
Martel A  Cross B 《Biomicrofluidics》2012,6(1):12813-128137
Artificial membranes, as a controllable environment, are an essential tool to study membrane proteins. Electrophysiology provides information about the ion transport mechanism across a membrane at the single-protein level. Unfortunately, high-throughput studies and screening are not accessible to electrophysiology because it is a set of not automated and technically delicate methods. Therefore, it is necessary to automate and parallelize electrophysiology measurement in artificial membranes. Here, we present a first step toward this goal: the fabrication and characterization of a microfluidic device integrating electrophysiology measurements and the handling of an artificial membrane which includes its formation, its displacement and the separation of its leaflets using electrowetting actuation of sub-μL droplets. To validate this device, we recorded the insertion of a model porin, α-hemolysin.  相似文献   

7.
Fluid shear stress (FSS) plays a critical role in regulating endothelium function and maintaining vascular homeostasis. Current microfluidic devices for studying FSS effects on cells either separate high shear stress zone and low shear stress zone into different culturing chambers, or arranging the zones serially along the flow direction, which complicates subsequent data interpretation. In this paper, we report a diamond shaped microfluidic shear device where the high shear stress zone and the low shear stress zone are arranged in parallel within one culturing chamber. Since the zones with different shear stress magnitudes are aligned normal to the flow direction, the cells in one stress group are not substantially affected by the flow-induced cytokine/chemokine releases by cells in the other group. Cell loading experiments using human umbilical vein endothelial cells show that the device is able to reveal stress magnitude-dependent and loading duration-dependent cell responses. The co-existence of shear stress zones with varied magnitudes within the same culturing chamber not only ensures that all the cells are subject to the identical culturing conditions, but also allows the resemblance of the differential shear stress pattern in natural arterial conditions. The device is expected to provide a new solution for studying the effects of heterogeneous hemodynamic patterns in the onset and progression of various vascular diseases.  相似文献   

8.
【目的/意义】从开放政府数据主题的多个政策文本的语义挖掘出发,发现多个政策文本内容间的语义关 系,探索能降低人工干预,实现多政策文本协同性自动化分析的方法。【方法/过程】利用数据挖掘的关联规则算法 对经过预处理的开放政府数据政策文本进行语义挖掘,按照得到的有效强关联分析多政策文本间的协同性。【结 果/结论】以开放政府数据主题的多个政策文本为研究对象,确定置信度为 0.7,提升度大于 3时得到的有效强关联 规则数量较稳定;经过不同层次的政策文本关联规则分析,可以得到与人工分析基本吻合的结论,验证了该方法可 以应用于多政策文本语义协同性的定量研究。【创新/局限】采用数据挖掘中的关联规则算法完成数据政策多文本 的协同性知识推理研究,有效的实现了语义自动化计算的问题。实验中政策词表的完整性、数据预处理过程、参数 设定等环节都会对实验结果准确性有影响,需进一步降低人工干预影响。  相似文献   

9.
热休克蛋白的研究进展及其应用   总被引:7,自引:0,他引:7  
靳远祥  陈玉银 《科技通报》2002,18(2):157-163
在不利的环境中,各种有机体都有其共同对应的分子反应,即正常基因的表达抑制和一组特殊基因-热休克基因的激活和表达,并导致热休克蛋白的大量产生。热休克蛋白主要作为分了伴侣而参与蛋白的折叠、转运及组装等过程,并能恢复或加速清除细胞内已变性的蛋白质而稳定细胞结构,使细胞产生热耐受。随着对热休克蛋白研究的不断深入,其在生物工程、医学和遗传育种等方面的应用前景十分广阔。  相似文献   

10.
Malaria infection is known to cause severe hemolysis due to production of abnormal RBCs and enhanced RBC destruction through apoptosis. Infected RBC lysis exposes uninfected RBC to the large amount of pro-oxidant molecules such as methemoglobin. Methemoglobin (MetHb) exposure dose dependently makes RBCs susceptible to osmotic stress and causes hemolysis. MetHb mediated oxidative stress in RBC correlated well with osmotic fragility and hemolysis. Interestingly, a reactive oxygen species (ROS) spike at 15 min was responsible for the observed effects on RBC cells. Two natural antioxidants N-acetyl cysteine and mannitol protected the RBC from MetHb-mediated defects, which clearly indicated involvement of oxidative stress in the process. MetHb due to its pseudo-peroxidase activity produces ROS in the external microenvironment. Therefore, classical peroxidase inhibitors were tested to probe peroxidase activity mediated ROS production with defects in RBCs. Clotrimazole (CLT), which irreversibly inactivates the MetHb (CLT-MetHb) and abolishes peroxidase activity, did not produce significant ROS outside RBC and was inefficient to cause osmotic fragility and hemolysis. Hence, initiating a chain reaction, MetHb released from ruptured RBC produces significant ROS in the external microenvironment to make RBC membrane leaky and enhanced hemolysis. Together data presented in the current work explored the role of MetHb in accelerated humorless during malaria which could be responsible for severe outcomes of pathological disorders.  相似文献   

11.
BackgroundIn industrial yeasts, selection and breeding for resistance to multiple stresses is a focus of current research. The objective of this study was to investigate the tolerance to multiple stresses of Saccharomyces cerevisiae obtained through an adaptive laboratory evolution strategy involving a repeated liquid nitrogen freeze–thaw process coupled with multi-stress shock selection. We also assessed the related resistance mechanisms and very high-gravity (VHG) bioethanol production of this strain.ResultsElite S. cerevisiae strain YF10-5, exhibiting improved VHG fermentation capacity and stress resistance to osmotic pressure and ethanol, was isolated following ten consecutive rounds of liquid nitrogen freeze–thaw treatment followed by plate screening under osmotic and ethanol stress. The ethanol yield of YF10-5 was 16% higher than that of the parent strain during 35% (w/v) glucose fermentation. Furthermore, there was upregulation of three genes (HSP26, HSP30, and HSP104) encoding heat-shock proteins involved in the stress response, one gene (TPS1) involved in the synthesis of trehalose, and three genes (ADH1, HXK1, and PFK1) involved in ethanol metabolism and intracellular trehalose accumulation in YF10-5 yeast cells, indicating increased stress tolerance and fermentative capacity. YF10-5 also showed excellent fermentation performance during the simultaneous saccharification and fermentation of VHG sweet potato mash, producing 13.40% (w/v) ethanol, which corresponded to 93.95% of the theoretical ethanol yield.ConclusionsA multiple-stress-tolerant yeast clone was obtained using adaptive evolution by a freeze–thaw method coupled with stress shock selection. The selected robust yeast strain exhibits potential for bioethanol production through VHG fermentation.How to cite: Zhang Q, Jin Y, Fang Y, et al. Adaptive evolution and selection of stress-resistant Saccharomyces cerevisiae for very high gravity bioethanol fermentation. Electron J Biotechnol 2019;41. https://doi.org/10.1016/j.ejbt.2019.06.003  相似文献   

12.
张毓敏 《科教文汇》2011,(36):85-86
最近十几年来,分子器件由于其诱人的应用前景已经成为了国际科技界竞争最为激烈的研究领域之一,这也使得分子器件领域在过去短短的时间里取得了很多优秀的研究成果。本文主要对分子器件的概念,以及分子器件研究中一些重要的实验和理论进展进行了简要的概括。  相似文献   

13.
Cryopreservation of human red blood cells (RBCs) in the presence of 40% glycerol allows a shelf-life of 10 years, as opposed to only 6 weeks for refrigerated RBCs. Nonetheless, cryopreserved blood is rarely used in clinical therapy, in part because of the requirement for a time-consuming (∼1 h) post-thaw wash process to remove glycerol before the product can be used for transfusion. The current deglycerolization process involves a series of saline washes in an automated centrifuge, which gradually removes glycerol from the cells in order to prevent osmotic damage. We recently demonstrated that glycerol can be extracted in as little as 3 min without excessive osmotic damage if the composition of the extracellular solution is precisely controlled. Here, we explore the potential for carrying out rapid glycerol extraction using a membrane-based microfluidic device, with the ultimate goal of enabling inline washing of cryopreserved blood. To assist in experimental design and device optimization, we developed a mass transfer model that allows prediction of glycerol removal, as well as the resulting cell volume changes. Experimental measurements of solution composition and hemolysis at the device outlet are in reasonable agreement with model predictions, and our results demonstrate that it is possible to reduce the glycerol concentration by more than 50% in a single device without excessive hemolysis. Based on these promising results, we present a design for a multistage process that is predicted to safely remove glycerol from cryopreserved blood in less than 3 min.  相似文献   

14.
小型单位有着其自身的特点,注册会计师应根据这些特点制定和实施相应的审计程序以形成正确的审计结论。分析了注册会计师在对小型单位审计时应特别关注的相关问题,以达到降低审计风险的目的。  相似文献   

15.
当塑料排水板手工插板枪超过5米时,由于单根长度太长携带很不方便。此外,在插板过程中,插板枪晃动等问题明显影响使用。本文针对一种新型的塑料排水板可伸缩式两段式插板枪,设计了其接头机械结构,分析了其接头的摩擦特性和受力状态,并通过现场插板实验验证了这种应用在大深度场合的两段式插板枪的功能和效率。  相似文献   

16.
We present a low cost microfluidic chip integrating 3D micro-chambers for the capture and the analysis of cells. This device has a simple design and a small footprint. It allows the implementation of standard biological protocols in a chip format with low volume consumption. The manufacturing process relies on hot-embossing of cyclo olefin copolymer, allowing the development of a low cost and robust device. A 3D design of microchannels was used to induce high flow velocity contrasts in the device and provide a selective immobilization. In narrow distribution channels, the liquid velocity induces a shear stress that overcomes adhesion forces and prevents cell immobilization or clogging. In large 3D chambers, the liquid velocity drops down below the threshold for cell attachment. The devices can be operated in a large range of input pressures and can even be handled manually using simple syringe or micropipette. Even at high flow injection rates, the 3D structures protect the captured cell from shear stress. To validate the performances of our device, we implemented immuno-fluorescence labeling and Fluorescence in Situ Hybridization (FISH) analysis on cancer cell lines and on a patient pleural effusion sample. FISH is a Food and Drug Administration approved cancer diagnostic technique that provides quantitative information about gene and chromosome aberration at the single cell level. It is usually considered as a long and fastidious test in medical diagnosis. This process can be easily implanted in our platform, and high resolution fluorescence imaging can be performed with reduced time and computer intensiveness. These results demonstrate the potential of this chip as a low cost, robust, and versatile tool adapted to complex and demanding protocols for medical diagnosis.  相似文献   

17.
涂层混凝土在应力腐蚀条件下氯离子渗透性能的变化是一个新的课题。本文利用自行设计的加载设备对混凝土保护涂层及涂层涂刷次数等不同情况下的氯离子渗透性进行了研究。研究表明,保护涂层减小了混凝土在应力腐蚀环境作用下抗弯强度的损失。同时也提高了混凝土的抗氯离子渗透性,而且随着保护涂层刷涂次数的增加这种优越性越明显。  相似文献   

18.
当塑料排水板手工插板枪超过5米时,由于单根长度太长携带很不方便。此外,在插板过程中,插板枪晃动等问题明显影响使用。本文设计了一种新型的塑料排水板可伸缩式插板枪结构,能在数秒钟内将两段式的插板枪便捷省力地组合成一根满足强度和刚度要求的单根插板枪并同样方便地拆卸为收缩式结构。所设计的可伸缩式插板枪在指定的栽荷作用下,由有限元分析软件计算出的各个零件最大应力均小于相对应材料的屈服极限值。不会产生永久性的变形。  相似文献   

19.
众创空间在高速发展的同时建设水平参差不齐,孵化效果相去甚远。本文选取了广州市的众创空间作为调研对象,调研众创空间的创业孵化模式及其效果,比较不同孵化模式的优劣。研究发现,孵化模式优劣的影响因素主要有三个方面:一是投入孵化要素的侧重点;二是众创空间对孵化模式的执行力度;三是孵化模式与孵化企业相互适应程度。从优化孵化模式出发,给众创空间的发展提出三点建议:一是众创空间应结合自身、行业、地区情况设计孵化模式;二是围绕孵化模式建立灵活的孵化机制;三是积极拓展投融资渠道。  相似文献   

20.
Implantable drug delivery systems can provide long-term reliability, controllability, and biocompatibility, and have been used in many applications, including cancer pain and non-malignant pain treatment. However, many of the available systems are limited to zero-order, inconsistent, or single burst event drug release. To address these limitations, we demonstrate prototypes of a remotely operated drug delivery device that offers controllability of drug release profiles, using osmotic pumping as a pressure source and magnetically triggered membranes as switchable on-demand valves. The membranes are made of either ethyl cellulose, or the proposed stronger cellulose acetate polymer, mixed with thermosensitive poly(N-isopropylacrylamide) hydrogel and superparamagnetic iron oxide particles. The prototype devices'' drug diffusion rates are on the order of 0.5–2 μg/h for higher release rate designs, and 12–40 ng/h for lower release rates, with maximum release ratios of 4.2 and 3.2, respectively. The devices exhibit increased drug delivery rates with higher osmotic pumping rates or with magnetically increased membrane porosity. Furthermore, by vapor deposition of a cyanoacrylate layer, a drastic reduction of the drug delivery rate from micrograms down to tens of nanograms per hour is achieved. By utilizing magnetic membranes as the valve-control mechanism, triggered remotely by means of induction heating, the demonstrated drug delivery devices benefit from having the power source external to the system, eliminating the need for a battery. These designs multiply the potential approaches towards increasing the on-demand controllability and customizability of drug delivery profiles in the expanding field of implantable drug delivery systems, with the future possibility of remotely controlling the pressure source.  相似文献   

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