Name: enhancement of the initial growth rate of agricultural plants by using static magnetic fields
Uploaded: 2016-07-08T13:00:00.000+00:00
Duration: 5 min 17 s
Description: Watch this Scientific Journal Video about Enhancement of the Initial Growth Rate of Agricultural Plants by Using Static Magnetic Fields at JoVE.com

This study received supported from the National Research Foundation of Korea (NRF) (2011-0012728). A poster presenting this study was awarded the Best Poster Award by the Korean Society of Applied Biological Sciences (KSABC).

1.初始设置<ol><li>农业植物物种<ol><li>用凤仙花（ 凤仙花），水菜（ 白菜变种粳稻 ），小松菜（ 白菜变种perviridis）和Mescluns（ 独行菜豌豆 ）种子。 注: 凤仙花 （凤仙花或玫瑰苦瓜）是印度本土的物种;一些成员也设在缅甸。小松（ 芜菁变种perviridis或小松 ）是相同种类的共同萝卜的变体。独行菜（ 独行菜豌豆 ）是一种药草被分类学相关的豆瓣和芥菜。他们有相似的味道和气味，为此，他们是商业利用5,7。 </li></ol></li><li>植物培养<ol><li>文化凤仙花（ 凤仙花），水菜（ 白菜变种粳稻 ），小松菜（ 白菜变种perviridis）和MesclunS（ 独行菜豌豆 ）种子在100毫米直径（100 PI）培养皿。保证一个板只包含一种类型的品种。 </li><li>对于培养条件，将种子在纤维素毛巾。浸泡毛巾和种子三重蒸馏水。测量和确认实验室室内RT是18-25°C，湿度与范围65-75％（请查看第3.1.2节）。 </li><li>为10±1种子凤仙花的，水菜的50±10种子，330±20种子小松菜的，和380±20 Mescluns的种子的种子数，培养。如使用18-25℃测量相同的条件下，用湿度范围65％-75％（请查阅2.1.1节）。 注:所有实验均在与调节的湿度和温度范围在实验室室内条件下进行的。湿度和温度不是静态的，而是提供了磁铁治疗组和对照相同的条件。 </li></ol></li></ol> 2.四大农业植物文化<ol><li>实验步骤<ol><li>遵循控制和磁体应用组的植物和文化条件的物种，第1.2.3节）。 </li><li>在100 PI菜为凤仙花底部的应用1750±350高斯（10000高斯= 1特斯拉）的三个磁体。在申请中，确保三个磁体不与种子直接接触，并且由培养皿的塑料底部是分开的。种子和磁铁之间的直接距离应为2-4毫米。申请的磁体168小时（7天）四农业植物。 </li><li>以下相同2.1.2所有步骤），在凤仙花培养板的底部运用两个磁体，一个（朝向Ñ侧上方）上的顶部和其他磁体（面对S端向上）。 注:极点在凤仙花不同的应用。然而，极取向不被认为是在本研究中为生长改变的一个关键因素，因为所有的环境是除了方向相同的磁通。 N和S极申请凤仙花的目的是要看到在字段，其中极取向可能是难以管理使用它的实际能力。 </li></ol></li></ol> 3.凤仙花的微管蛋白染色<ol><li>磁体应用程序与管制的光照条件下<ol><li>将（N极朝上）100毫米的钢板底48小时，在步骤1.2.2使用条件的两个磁铁。 注:对于光的修改中，培养皿置于培养箱中一塑料架。用孵化器的光的拦截和保持温度在25℃在黑暗的环境中48小时。最终，这种情况不是本实验由于生长长高变使用。 </li></ol></li><li>植物染色<ol><li>修复整个凤仙花SPP双花植物（包括茎和根）生长在相同的条件，在4％多聚甲醛和0.1M的步骤3.1.2）磷酸盐缓冲液（pH7.4）15分钟。 </li><li>除去凤仙花样本，并在封闭缓冲液（在PBS中2％马血清/ 1％牛血清白蛋白/ 0.1％的Triton X-100，pH值7.5）浸泡2小时。用PBS浸泡15分钟，洗净凤仙样本。 </li><li>对于双免疫染色，孵育样品与第一抗体，抗α微管蛋白（1:1000），O / N在4℃。 </li><li>取出样品并10分钟洗一次，用PBS浸泡样品。使用FITC-缀合的抗小鼠IgG（1:400）作为第二抗体，并在25℃孵育2小时。 </li><li>浸入在PBS样品并覆盖滑整个样品在24孔板的底部。获得使用传统的荧光显微镜来观察微管方向影像（λ= 550纳米，放大到100X，200X和400X）。 注:在这种情况下，磁铁治疗组（n = 10）和对照组（11例）被确认为凤仙花（ 凤仙花 ）只在非黑暗条件下生长。 </LI&gt; </ol></li></ol> 4.数据收集方法<ol><li>四农业植物生长的时间推移创建<ol><li>拍摄植物以10分钟的间隔，通过设置快门自动（这可以在任何数字照相机来完成）。设定光圈至F 3.2和ISO值400。 </li><li>收集700-900图片为7-10天。相机与电线连接，因为电池会耗尽。 </li><li>通过点击和电影制作软件流线以下按时间顺序下探每一张图片（见材料和​​设备表）拖动图片。穿上它在0.045-0.05秒，每个持续时间相等的流线成片共有30-40秒。检查，以便有在时间顺序选择每一个画面无暗差距。 </li><li> 4.1.3步后，点击软件播放按钮，以确保编译-电影成30-40秒的时间推移视频幻灯片，然后点击渲染并保存到文件.mpeg或.AVI格式。对于大小马rkers，使用加拿大季度，美国的竹篙，和一厘米的统治者照片的一面。 </li><li>执行t检验和箱形图进行统计分析11,12。 注:五个数摘要组被用于计算下限值（L）的值作为Q 1 - [1.5×（Q3 - Q 1）]和上限（H）的值作为Q 3 + 1.5×（Q3 - Q 1） ]。这种做法被纳入步1.2.2长度的数据收集11。在L和H值显示在T-分布图的99％的区域，这意味着在此范围外观察到的数据点可以被认为是异常值。 -测试被用来分析在苗木12的高度差异箱图和学生的t。 </li></ol></li></ol>

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The Authors have nothing to disclose.

在所有的情况下，磁体应该培养皿下应用。本研究对种子的增长速度磁场的影响，若干农产品品种，重点凤仙花农业植物的代表。例如，微管蛋白染色上凤仙花执行以评估根在分子水平的变化和茎骨架微型结构表明在长度扩散磁场的影响。无论是磁石的N极和S极长期（7-10 d）利用凤仙花随访研究中的应用。其他三个品种，水菜（ 白菜变种粳稻 ），小松菜（ 白菜变种perviridis）和Mesclun（ 独行菜豌豆 ），用N极磁铁导向治疗。这是为了进一步验证该静磁场本身，而不是磁极，在初始生长增强的主要因素。此外，增加物种数量提供支持在农业植物体来源的初始生长的促进更广泛的适用性。 许多因素，如营养，湿度，温度，光，可能影响植物生长3的速率。每一种举行跨治疗不变。营养补充品被排除只在三蒸水培养。我们首先控制了光实验黑暗条件下的孵化器最初进行的凤仙花。在昏暗的环境下增长模式从那些在光环境不同。因此，我们的光线条件（利用光的相同数额各实验组）下进行后续实验。对于微管蛋白染色的凤仙花被控制的条件（三蒸水，温度18-25℃，湿度为65％-75％）下生长。 7-10-D的随访研究的其他实验有相同的"空条件:无营养"的条件，那些使用嘎RDEN香脂（三重的蒸馏水，温度18-25℃，湿度65％-75％）。相对于磁体的应用中，我们使用的策略，其中我们定量增加种类数量和磁铁施加的持续时间，以进一步调查磁场是否对农业植物不限于某些种类的通用生长促进效果。用凤仙花（ 凤仙花），水菜（ 白菜变种粳稻 ），小松菜（ 白菜变种perviridis）和Mescluns（ 独行菜大蒜 ）这个想法进行了研究。 这种现象的分子基础部分由微管蛋白染色实验9-11阐明，但需要对实际使用进一步调查。精确的磁应用程序可以在由于磁铁本身的侵蚀潮湿环境的限制。磁场物理提高农业植物的生长。然而，THIs不证明营养成分也增加。的植物的化学成分进一步分析，以确定使用磁场是否具有相似的肥料的效果来进行。这也可以在其中提供养分的环境中，以及在营养空条件下，使用在本研究中采用了蒸馏水进行评价。除了质量（类型，强度等）和施加的磁体数量，成本可以是复杂这样的应用的另一个问题。它可以是昂贵的，以一整个作物田间应用众多磁铁。 我们的研究结果表明，静磁场的应用加速发芽率和一些栽培植物物种，最初的增长速度。这些结果表明，静态磁场具有对植物生长，尤其是植物的发芽6和根生长7显著效果。先前的研究已经表明，最大发芽率分别高20％时的磁场的频率为大约10赫兹5-6。期间仅4天的磁场的应用，茎和根长增加。经受磁场治疗（N = 10）组具有较高的1.4倍增长速度比在总共8天（P &lt;0.0005）的做了对照组（n = 11）。这个速度比以前的研究，使用脉冲磁场6-9发现高出20％。 根据这些发现，基因表达和调节也应为澄清的背后观察应答的潜在机制未来的实验研究，以磁场10。我们的研究结果表明，磁场的应用可以提高作物的增长速度，这可能有助于解决全球粮食和贫困问题。此外，磁场的应用可能是有用减少使用化学肥料。

发芽是导致幼苗1的形成一个植物的生长。在一定条件下，种子萌发开始和胚胎组织恢复生长。它始于水化到种子以激活酶发芽。种子可诱导体外萌发（在培养皿或试管）中1,2。 静态磁场是特种兵导致由洛伦兹力3,4方式离子电荷的分子的运动。形成洛伦兹力时在磁场下的离子化或带电物体移动。每一材料与组成电子和质子的原子形成。当磁场成为目前，无论是静态的或交替，它会影响带电材料的移动。这也适用于植物和水分子，从而影响细胞内分子状态。在先前的研究中，使用电磁线圈产生脉冲磁场，和&#39;小松菜&#39;植物被选择作为研究对象5。在本研究中，磁铁产生静磁场被用来得到类似但不同的作用效果的洛伦兹力的膨胀研究。 磁场的频率，而不是它的极性，是植物发芽的关键因素。先前的研究已经表明，最大发芽率比对照组高20％时的磁场的频率约为10赫兹。当以逆行方式除去字段，增速受损5。静磁场对初始生长6-8相当大的影响，主要是对发芽6和根系生长7。 在本研究中，我们使用的静态磁铁来检查通过使用磁场调节农业植物生长的可能性。特别是，我们的目的至determine磁场应用的特定的条件是否能增加的生长速率，以更高的水平比在文献中提到的那些。此外，如果可以使用磁场来成功地提高植物的初始发芽，可避免使用化学肥料。

<table><tbody><tr><td>Static magnets</td><td>JIM</td><td></td><td>2000Gauss</td></tr><tr><td>2% horse serum/1% bovine serum albumin/0.1% Triton X-100</td><td>Sigma-Aldrich</td><td>Merged with 55514</td><td>Blocking buffer</td></tr><tr><td>Primary antibody</td><td>Santa Cruz Biotechnology</td><td>sc-8035</td><td>a-Tubulin</td></tr><tr><td>Secondary antibody</td><td>Santa Cruz Biotechnology</td><td>sc-2010</td><td>FITC-conjugated anti-mouse IgG</td></tr><tr><td>time lapse photographic techniques</td><td>Manually controlled</td><td></td><td>ISO value 400 &amp;amp; aperture F 3.2</td></tr><tr><td>Sony Vegas Pro 13.0</td><td>Sony</td><td></td><td></td></tr></tbody></table>

enhancement of the initial growth rate of agricultural plants by using static magnetic fields

电子器件和高压电线诱导磁场。 1,300-2,500高斯（0.2特斯拉）的磁场加到含有凤仙花（ 凤仙花），水菜（ 芜青变种粳稻 ），小松菜的种子皮氏培养皿（ 芜菁变种perviridis）和Mescluns（ 独行菜豌豆 ）。我们采用培养皿下磁铁。在应用程序的4天，我们观察到，茎和根长度增加。在总共8天（P &lt;0.0005）的对照组（n = 11）相比，经受磁场的治疗（10）的组显示出1.4倍的速度增长速度。这速率比在以前的研究中报道的高20％。微管蛋白复合线没有连接点，但在磁铁的应用程序出现连接点。这显示了从控制，这意味着异常安排完全不同。不过，确切原因尚不清楚。这些水库施加磁体的生长增强的ULTS表明，它能够提高生长速度，提高生产效率，或通过施加静态磁场控制植物发芽的速度。另外，磁场可能引起在植物细胞中的生理变化，并且可以诱导生长。因此，刺激与磁场可具有类似于化学肥料，这意味着，肥料的使用可避免的可能影响。

微管蛋白染色显示分散或在磁体的存在下与对照相比（ 图2）生长的植物变薄结构。此外，农业植物，包括小松菜（ 芜菁变种perviridis）和Mescluns（ 独行菜豌豆 ）7-一天的时间推移研究表明，一磁铁衍生静磁场增加这些植物的初始生长（ 图3）。 这些结果表明，暴露于磁场的组中有显着的增长变化（ 图1）。在黑暗的环境中生长的植物没有表现出任何差异，这表明只有光存在的条件是适用在7天的时间推移的实验。三个具有代表性的农业植物在该研究中使用，但也已经使用更多的植物。农作物和其他植物可以使用进行调查相同的协议。在以往的研究中，生长速度提高了20％，而本结果表明一个1.4倍的增加，这是40％。因此，利用静磁场的磁体的应用比交变磁场脉冲的施加更加有效。 确定磁场的效果可以复杂，因为与电荷的任何分子结构可影响3,4。静磁场似乎影响凤仙花的初始生长速率在培养的纤维素塔组织。值有统计学显著，是在控制的增长速度约为1.4倍。微管蛋白是细胞伸长和增长9中保持种植结构至关重要。 <img alt="图1" src="/files/ftp_upload/53967/53967fig1.jpg" /> 图1.增长凤仙花的。 </str翁&gt;（A）凤仙花与黑暗条件下的边际下的静磁场处理的增长;然而，当暴露于光（示出的唯一代表图像）的植物生长得更快。 （B）当暴露在光线下，第3天，高度差异有统计学显著（P &lt;0.01，双侧t检验）。 （ 三 ）每个单株的高度是到晚上7个交易日走高（**:上限标准误差的测量）。黑暗条件下没有引起任何差异，表明该磁场的影响可能与激素有关。 <a href="https://www.jove.com/files/ftp_upload/53967/53967fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/53967/53967fig2.jpg" /> 图2. 凤仙花和微管蛋白染色增加磁场的申请后，凤仙花的增长速度。（A）凤仙花显示，微管蛋白结构的分散分布在施加一个磁场。这一发现表明，生长拦阻蛋白质结构如微管蛋白（和可能的肌动蛋白）由静磁场的影响。 （B）的平均增长速度为1.4倍，比对照高，平均高度为与磁场治疗组高。 <a href="https://www.jove.com/files/ftp_upload/53967/53967fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/53967/53967fig3.jpg" /> 图3.磁场促进Mescluns的（ 独行菜大蒜，前面）和小松菜的生长（ 芜菁变种perviridis）种子SUP执行完成培养皿用1750±350高斯的磁场处理和7天观察到10分钟的时间推移的时间间隔。时间推移视频切割成每11小时15片段。 <a href="https://www.jove.com/files/ftp_upload/53967/53967fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="视频1帧" src="/files/ftp_upload/53967/53967video1.jpg" /> 补充视频1:凤仙花（ 凤仙花 ）一个凤仙花（ 凤仙花 ）种子补充培养皿与1750±350高斯的磁场处理，然后观察7天为10的延时间隔的成长时光倒流分钟。该视频被重新安排到一个30分钟电影。 <a href="https://www.jove.com/files/ftp_upload/53967/timelapse1.mp4">请点击此处观看该视频。 （右键点击下载）。</a> 补充视频2:在一式三份培养3植物的生长的比较水菜（芜菁变种粳稻 ），小松菜（芜菁变种perviridis）和Mesclun（ 独行菜豌豆 ）种子示于一个直径为100毫米（100圆周） 培养皿。在相同条件下对这些凤仙花的，三 ​​个品种分别进行评估，结果发现，该磁铁的效果在农业植物广泛遵守。 <a href="https://www.jove.com/files/ftp_upload/53967/timelapse2.avi">请点击此处观看该视频。 （右键点击下载）。</a>

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Enhancement of the Initial Growth Rate of Agricultural Plants by Using Static Magnetic Fields

The goal of this protocol is to demonstrate the acceleration of the initial growth rate of plants by applying static magnetic fields with no external energy.

采用静态磁场农业植物初始增长率提高

Electronic devices and high-voltage wires induce magnetic fields. A magnetic field of 1,300-2,500 Gauss (0.2 Tesla) was applied to Petri dishes containing ...

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Magnetic Forces and Fields

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13.2K 次观看.  Yonsei University. The goal of this protocol is to demonstrate the acceleration of the initial growth rate of plants by applying static magnetic fields with no external energy.

视频: Enhancement of the Initial Growth Rate of Agricultural Plants by Using Static Magnetic Fields

Geomagnetic Field (Gmf) and Plant Evolution: Investigating the Effects of Gmf Reversal on Arabidopsis thaliana Development and Gene Expression

One of the most stimulating observations in plant evolution is a correlation between the occurrence of geomagnetic field (GMF) reversals (or excursions) and the moment of the radiation of Angiosperms. This led to the hypothesis that alterations in GMF polarity may play a role in plant evolution. Here, we describe a method to test this hypothesis by exposing Arabidopsis thaliana to artificially reversed GMF conditions. We used a three-axis magnetometer and the collected data were used to calculate the magnitude of the GMF. Three DC power supplies were connected to three Helmholtz coil pairs and were controlled by a computer to alter the GMF conditions. Plants grown in Petri plates were exposed to both normal and reversed GMF conditions. Sham exposure experiments were also performed. Exposed plants were photographed during the experiment and images were analyzed to calculate root length and leaf areas. Arabidopsis total RNA was extracted and Quantitative Real Time-PCR (qPCR) analyses were performed on gene expression of CRUCIFERIN 3 (CRU3), copper transport protein1 (COTP1), Redox Responsive Transcription Factor1 (RRTF1), Fe Superoxide Dismutase 1, (FSD1), Catalase3 (CAT3), Thylakoidal Ascorbate Peroxidase (TAPX), a cytosolic Ascorbate Peroxidase1 (APX1), and NADPH/respiratory burst oxidase protein D (RbohD). Four different reference genes were analysed to normalize the results of the qPCR. The best of the four genes was selected and the most stable gene for normalization was used. Our data show for the first time that reversing the GMF polarity using triaxial coils has significant effects on plant growth and gene expression. This supports the hypothesis that GMF reversal contributes to inducing changes in plant development that might justify a higher selective pressure, eventually leading to plant evolution.

Geomagnetic Field Gmf and Plant Evolution: Investigating the Effects of Gmf Reversal on Arabidopsis thaliana Development and Gene Expression

This protocol describes a method to test the hypothesis that the reversal of the effective geomagnetic field (GMF) induces differential gene expression and alters the morphology of Arabidopsis thaliana. A triaxial octagonal system of Helmholtz coil-pairs was used to artificially generate reversed GMF conditions in the plant growth chamber.

地磁场（GMF）和植物进化:调查逆转基因食品对影响<em&gt;拟南芥</em&gt;开发与基因表达

One of the most stimulating observations in plant evolution is a correlation between the occurrence of geomagnetic field (GMF) reversals (or excursions) ...

科研

发育生物学

Robotic Sensing and Stimuli Provision for Guided Plant Growth

Robot systems are actively researched for manipulation of natural plants, typically restricted to agricultural automation activities such as harvest, irrigation, and mechanical weed control. Extending this research, we introduce here a novel methodology to manipulate the directional growth of plants via their natural mechanisms for signaling and hormone distribution. An effective methodology of robotic stimuli provision can open up possibilities for new experimentation with later developmental phases in plants, or for new biotechnology applications such as shaping plants for green walls. Interaction with plants presents several robotic challenges, including short-range sensing of small and variable plant organs, and the controlled actuation of plant responses that are impacted by the environment in addition to the provided stimuli. In order to steer plant growth, we develop a group of immobile robots with sensors to detect the proximity of growing tips, and with diodes to provide light stimuli that actuate phototropism. The robots are tested with the climbing common bean, Phaseolus vulgaris, in experiments having durations up to five weeks in a controlled environment. With robots sequentially emitting blue light-peak emission at wavelength 465 nm-plant growth is successfully steered through successive binary decisions along mechanical supports to reach target positions. Growth patterns are tested in a setup up to 180 cm in height, with plant stems grown up to roughly 250 cm in cumulative length over a period of approximately seven weeks. The robots coordinate themselves and operate fully autonomously. They detect approaching plant tips by infrared proximity sensors and communicate via radio to switch between blue light stimuli and dormant status, as required. Overall, the obtained results support the effectiveness of combining robot and plant experiment methodologies, for the study of potentially complex interactions between natural and engineered autonomous systems.

Distributed robot nodes provide sequences of blue light stimuli to steer the growth trajectories of climbing plants. By activating natural phototropism, the robots guide the plants through binary left-right decisions, growing them into predefined patterns that by contrast are not possible when the robots are dormant.

机器人传感和刺激为引导植物生长提供

Robot systems are actively researched for manipulation of natural plants, typically restricted to agricultural automation activities such as harvest, ...

工程学

Control and Disposal of Invasive Japanese Knotweed Reynoutria japonica Houtt. Using Microwave Treatment

The study aims to assess the effectiveness of microwave treatment (MWT) at a frequency of 2.45 GHz and a power of 800 W to control Japanese knotweed (Reynoutria japonica Houtt.) using a self-propelled device that was built in the in-house facility. The MWT was applied in the field population of knotweed in July 2022. First, plants were mechanically moved from the area of 1 m2, and next, the cut shoots around 4 cm high were microwave-treated for 25 min, 20 min, and 15 min. The control treatments were: 1) only cut plants and 2) rhizomes dug out to 30 cm deep. The effectiveness of the microwave treatments was observed for the next 11 months by counting the number of newly grown shoots. The results showed that a 25 min MWT was 100% effective in Japanese knotweed loss of vitality, while a 15 min MWT microwave treatment stimulated plant growth by around 50%, compared to controls. Rhizomes were dug out in a separate in vitro experiment for laboratory testing. The rhizomes were categorized by thickness and subjected to a 60 s MWT using a commercial microwave, after which their temperature and vitality were assessed. The temperature of rhizomes following MWT depended on their thickness. Those rhizomes that warmed to temperatures above 42 &#176;C were effectively destroyed. Summing up, the time plants are exposed to microwaves plays a major role in the effectiveness of this method. The longer the exposure to MWT, the better control. The thinner the rhizomes, the more effective the in vitro MWT rhizomes disposal.

This protocol provides a method for microwave control of Japanese knotweed in the field and disposal of dug-up rhizomes in laboratory conditions. The advantages and disadvantages of both methods are discussed. Future research directions are also suggested to optimize the use of microwaves for controlling Japanese knotweed.

The study aims to assess the effectiveness of microwave treatment (MWT) at a frequency of 2.45 GHz and a power of 800 W to control Japanese knotweed ...

环境科学

3D Magnetic Stem Cell Aggregation and Bioreactor Maturation for Cartilage Regeneration

Cartilage engineering remains a challenge due to the difficulties in creating an in vitro functional implant similar to the native tissue. An approach recently explored for the development of autologous replacements involves the differentiation of stem cells into chondrocytes. To initiate this chondrogenesis, a degree of compaction of the stem cells is required; hence, we demonstrated the feasibility of magnetically condensing cells, both within thick scaffolds and scaffold-free, using miniaturized magnetic field sources as cell attractors. This magnetic approach was also used to guide aggregate fusion and to build scaffold-free, organized, three-dimensional (3D) tissues several millimeters in size. In addition to having an enhanced size, the tissue formed by magnetic-driven fusion presented a significant increase in the expression of collagen II, and a similar trend was observed for aggrecan expression. As the native cartilage was subjected to forces that influenced its 3D structure, dynamic maturation was also performed. A bioreactor that provides mechanical stimuli was used to culture the magnetically seeded scaffolds over a 21-day period. Bioreactor maturation largely improved chondrogenesis into the cellularized scaffolds; the extracellular matrix obtained under these conditions was rich in collagen II and aggrecan. This work outlines the innovative potential of magnetic condensation of labeled stem cells and dynamic maturation in a bioreactor for improved chondrogenic differentiation, both scaffold-free and within polysaccharide scaffolds.

Chondrogenesis from stem cells requires fine tuning the culture conditions. Here, we present a magnetic approach for condensing cells, an essential step to initiate chondrogenesis. In addition, we show that dynamic maturation in a bioreactor applies mechanical stimulation to the cellular constructs and enhances cartilaginous extracellular matrix production.

3D磁干细胞聚集和生物反应器为成熟软骨再生

Cartilage engineering remains a challenge due to the difficulties in creating an in vitro functional implant similar to the native tissue. An approach ...

生物工程

External Excitation of Neurons Using Electric and Magnetic Fields in One- and Two-dimensional Cultures

A neuron will fire an action potential when its membrane potential exceeds a certain threshold. In typical activity of the brain, this occurs as a result of chemical inputs to its synapses. However, neurons can also be excited by an imposed electric field. In particular, recent clinical applications activate neurons by creating an electric field externally. It is therefore of interest to investigate how the neuron responds to the external field and what causes the action potential. Fortunately, precise and controlled application of an external electric field is possible for embryonic neuronal cells that are excised, dissociated and grown in cultures. This allows the investigation of these questions in a highly reproducible system.
In this paper some of the techniques used for controlled application of external electric field on neuronal cultures are reviewed. The networks can be either one dimensional, i.e. patterned in linear forms or allowed to grow on the whole plane of the substrate, and thus two dimensional. Furthermore, the excitation can be created by the direct application of electric field via electrodes immersed in the fluid (bath electrodes) or by inducing the electric field using the remote creation of magnetic pulses.

Neuronal cultures are a good model for studying emerging brain stimulation techniques via their effect on single neurons or a population of neurons. Presented here are different methods for stimulation of patterned neuronal cultures by an electric field produced directly by bath electrodes or induced by a time-varying magnetic field.

使用电动和一维和二维文化磁场神经元的外部激励

A neuron will fire an action potential when its membrane potential exceeds a certain threshold. In typical activity of the brain, this occurs as a result ...

神经科学

Electric and Magnetic Field Devices for Stimulation of Biological Tissues

Electric fields (EFs) and magnetic fields (MFs) have been widely used by tissue engineering to improve cell dynamics such as proliferation, migration, differentiation, morphology, and molecular synthesis. However, variables such stimuli strength and stimulation times need to be considered when stimulating either cells, tissues or scaffolds. Given that EFs and MFs vary according to cellular response, it remains unclear how to build devices that generate adequate biophysical stimuli to stimulate biological samples. In fact, there is a lack of evidence regarding the calculation and distribution when biophysical stimuli are applied. This protocol is focused on the design and manufacture of devices to generate EFs and MFs and implementation of a computational methodology to predict biophysical stimuli distribution inside and outside of biological samples. The EF device was composed of two parallel stainless-steel electrodes located at the top and bottom of biological cultures. Electrodes were connected to an oscillator to generate voltages (50, 100, 150 and 200 Vp-p) at 60 kHz. The MF device was composed of a coil, which was energized with a transformer to generate a current (1 A) and voltage (6 V) at 60 Hz. A polymethyl methacrylate support was built to locate the biological cultures in the middle of the coil. The computational simulation elucidated the homogeneous distribution of EFs and MFs inside and outside of biological tissues. This computational model is a promising tool that can modify parameters such as voltages, frequencies, tissue morphologies, well plate types, electrodes and coil size to estimate the EFs and MFs to achieve a cellular response.

This protocol describes the step-by-step process to build both electrical and magnetic stimulators used to stimulate biological tissues. The protocol includes a guideline to simulate computationally electric and magnetic fields and manufacture of stimulator devices.

用于刺激生物组织的电动和磁场设备

Electric fields (EFs) and magnetic fields (MFs) have been widely used by tissue engineering to improve cell dynamics such as proliferation, migration, ...

Optimization and Utilization of Agrobacterium-mediated Transient Protein Production in Nicotiana

Agrobacterium-mediated transient protein production in plants is a promising approach to produce vaccine antigens and therapeutic proteins within a short period of time. However, this technology is only just beginning to be applied to large-scale production as many technological obstacles to scale up are now being overcome. Here, we demonstrate a simple and reproducible method for industrial-scale transient protein production based on vacuum infiltration of Nicotiana plants with Agrobacteria carrying launch vectors. Optimization of Agrobacterium cultivation in AB medium allows direct dilution of the bacterial culture in Milli-Q water, simplifying the infiltration process. Among three tested species of Nicotiana, N. excelsiana (N. benthamiana &#215; N. excelsior) was selected as the most promising host due to the ease of infiltration, high level of reporter protein production, and about two-fold higher biomass production under controlled environmental conditions. Induction of Agrobacterium harboring pBID4-GFP (Tobacco mosaic virus-based) using chemicals such as acetosyringone and monosaccharide had no effect on the protein production level. Infiltrating plant under 50 to 100 mbar for 30 or 60 sec resulted in about 95% infiltration of plant leaf tissues. Infiltration with Agrobacterium laboratory strain GV3101 showed the highest protein production compared to Agrobacteria laboratory strains LBA4404 and C58C1 and wild-type Agrobacteria strains at6, at10, at77 and A4. Co-expression of a viral RNA silencing suppressor, p23 or p19, in N. benthamiana resulted in earlier accumulation and increased production (15-25%) of target protein (influenza virus hemagglutinin).

Optimization and Utilization of Agrobacterium-mediated Transient Protein Production in Nicotiana

Transient protein production in Nicotiana plants based on vacuum infiltration with Agrobacteria carrying launch vectors (Tobacco mosaic virus-based) is a rapid and economic approach to produce vaccine antigens and therapeutic proteins. We simplified the procedure and improved target accumulation by optimizing conditions of bacteria cultivation, selecting host species, and co-introducing RNA silencing suppressors.

优化和利用<em&gt;农杆菌</em&gt;介导的瞬态蛋白生产中<em&gt;烟草</em

Agrobacterium-mediated transient protein production in plants is a promising approach to produce vaccine antigens and therapeutic proteins within a short ...

生物学

Growing Magnetotactic Bacteria of the Genus Magnetospirillum: Strains MSR-1, AMB-1 and MS-1

Magnetotactic bacteria are Gram-negative, motile, mainly aquatic prokaryotes ubiquitous in freshwater and marine habitats. They are characterized by their ability to biomineralize magnetosomes, which are magnetic nanometer-sized crystals of magnetite (Fe3O4) or greigite (Fe3S4) surrounded by a lipid bilayer membrane, within their cytoplasm. For most known magnetotactic bacteria, magnetosomes are assembled in chains inside the cytoplasm, thereby conferring a permanent magnetic dipole moment to the cells and causing them to align passively with external magnetic fields. Because of these specific features, magnetotactic bacteria have a great potential for commercial and medical applications. However, most species are microaerophilic and have specific O2 concentration requirements, making them more difficult to grow routinely than many other bacteria such as Escherichia coli. Here we present detailed protocols for growing three of the most widely studied strains of magnetotactic bacteria, all belonging to the genus Magnetospirillum. These methods allow for precise control of the O2 concentration made available to the bacteria, in order to ensure that they grow normally and synthesize magnetosomes. Growing magnetotactic bacteria for further studies using these procedures does not require the experimentalist to be an expert in microbiology. The general methods presented in this article may also be used to isolate and culture other magnetotactic bacteria, although it is likely that growth media chemical composition will need to be modified.

Growing Magnetotactic Bacteria of the Genus Magnetospirillum: Strains MSR-1, AMB-1 and MS-1

We present a procedure for growing several strains of Magnetospirillum in two different types of growth media. Magnetospirillum gryphiswaldense strain MSR-1 is grown in both liquid and O2 concentration gradient semi-solid media while M. magneticum strain AMB-1 and M. magnetotacticum strain MS-1 are grown in liquid medium.

螺属的生长趋磁菌: 菌株 MSR-1、AMB-1 和 MS-1

Magnetotactic bacteria are Gram-negative, motile, mainly aquatic prokaryotes ubiquitous in freshwater and marine habitats. They are characterized by their ...

MRM Microcoil Performance Calibration and Usage Demonstrated on Medicago truncatula Roots at 22 T

This protocol describes a signal-to-noise ratio (SNR) calibration and sample preparation method for solenoidal microcoils combined with biological samples, designed for high-resolution magnetic resonance imaging (MRI), also referred to as MR microscopy (MRM). It may be used at pre-clinical MRI spectrometers, demonstrated on Medicago truncatula root samples. Microcoils increase sensitivity by matching the size of the RF resonator to the size of the sample of interest, thereby enabling higher image resolutions in a given data acquisition time. Due to the relatively simple design, solenoidal microcoils are straightforward and cheap to construct and can be easily adapted to the sample requirements. Systematically, we explain how to calibrate new or home-built microcoils, using a reference solution. The calibration steps include: pulse power determination using a nutation curve; estimation of RF-field homogeneity; and calculating a volume-normalized signal-to-noise ratio (SNR) using standard pulse sequences. Important steps in sample preparation for small biological samples are discussed, as well as possible mitigating factors such as magnetic susceptibility differences. The applications of an optimized solenoid coil are demonstrated by high-resolution (13 x 13 x 13 &#956;m3, 2.2 pL) 3D imaging of a root sample.

MRM Microcoil Performance Calibration and Usage Demonstrated on Medicago truncatula Roots at 22 T

A protocol to study biological tissue at high spatial resolution using ultra-high field magnetic resonance microscopy (MRM) using microcoils is presented. Step-by-step instructions are provided for characterizing the microcoils. Finally, optimization of imaging is demonstrated on plant roots.

MRM 微可可性能校准和使用在 22 T 的 Medicago 特伦卡图拉根上演示

This protocol describes a signal-to-noise ratio (SNR) calibration and sample preparation method for solenoidal microcoils combined with biological ...

Stimulation of Patterned Neuronal Cultures by a Magnetic Field

Source: Stern, S., et al., <a href="https://app.jove.com/v/54357">External Excitation of Neurons Using Electric and Magnetic Fields in One- and Two-dimensional Cultures.</a> J. Vis. Exp. (2017)
This video demonstrates a method for stimulating neuronal cultures grown in a patterned circular ring on a coverslip, using a time-varying magnetic field.

磁场刺激模式化神经元培养物

1. Preparation of coverslips for 1D cultures.

	Clean glass coverslips by immersion in a base piranha solution consisting of 75 mL DDW, 25 mL 25% ammonia ...

采用静态磁场农业植物初始增长率提高

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地磁场（GMF）和植物进化:调查逆转基因食品对影响