Name: 视频: The 2009 Lindau Nobel Laureate Meeting: Martin Chalfie, Chemistry 2008
Uploaded: 2010-02-10T18:45:00.000+00:00
Duration: 12 min 49 s
Description: 6.7K 次观看. 美国生物学家马丁Chalfie罗杰钱学森和下村修分享2008年诺贝尔化学奖的绿色荧光蛋白（GFP）的发现和发展。 Chalfie克隆的GFP编码序列，并表示它在这两个 E。大肠杆菌 C。线虫，第一次证明，没有其他的因素是GFP的发光。

<ol>
	<li>Chalfie, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=GFP:+Lighting+up+life.">GFP: Lighting up life.</a> PNAS. 106 (25), 10073-10080 (2009).</li><li>Chalfie, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Green+fluorescent+protein+as+a+marker+for+gene+expression.">Green fluorescent protein as a marker for gene expression.</a> Science. 263 (5148), 802-805 (1994).</li></ol>

the 2009 lindau nobel laureate meeting: martin chalfie, chemistry 2008

美国生物学家马丁Chalfie罗杰钱学森和下村修分享2008年诺贝尔化学奖的绿色荧光蛋白（GFP）的发现和发展。 Chalfie马丁出生于1947年在芝加哥和斯科基伊利诺伊长大。虽然他从青年时代在科学的兴趣 - 学习关于恐龙的行星和阅读书籍的名称 - ，他在生物科学的职业生涯之旅迂回。 Chalfie在高中时，喜欢他的AP化学课程，但他的其他科学课程没有对他的印象，他开始在哈佛不确定是什么，他想研究他的本科学业。最后，他选择生物化学专业，并在之间的大二和大三年夏天，他加入了克劳斯韦伯的实验室，开始了他的第一个真正的研究项目，研究的酶天冬氨酸transcarbamylase的活性部位。不幸的是，他在韦伯的实验室进行的实验工作，并Chalfie来，不是为他的研究结论。 于1969年毕业后，他雇用了作为一名教师哈姆登厅国家日在康涅狄格州，在学校他教了2年高中化学，代数，科学和社会科学。他第一年的教学后，他决定放弃研究再次尝试。他花了一个夏天在耶鲁大学在圣何塞Zadunaisky的实验室工作，研究氯离子运输中的青蛙视网膜。 Chalfie享受了很多这方面的经验，并获得了在他自己的科学能力的信心，他申请，在哈佛大学毕业，在那里他于1972年加入生理学系，师从鲍勃珀尔曼的去甲肾上腺素的合成和分泌的学校。他的兴趣在C 上工作线虫导致他后DOC与悉尼布伦纳在英国医学研究委员会剑桥分子生物学实验室在，英国，。 1982年，他在哥伦比亚大学提供的位置。 当Chalfie第一次在绿色荧光蛋白的研究研讨会于1989年由保罗布雷姆听到，他的实验室研究的发展和触摸敏感细胞的功能，在C有关的基因线虫 。他立刻成为线虫中表达荧光蛋白的想法感到非常兴奋，希望能找出表达的基因在活的有机体。当时，所有检查本地化的方法，如抗体染色或原位杂交，需要固定的组织或细胞，揭示了蛋白质，只能在固定时间点的位置。 1992年9月，从道格拉斯Prasher获得绿色荧光蛋白的DNA后，Chalfie问他旋转的学生，吉亚奥伊斯基兴，以表达绿色荧光蛋白在大肠杆菌中大肠杆菌 ，不知道，其他几个实验室也试图表达的蛋白质，但没有成功。 Chalfie和奥伊斯基兴用于PCR扩增只有GFP的编码序列，它们放置在一个表达载体，并在大肠杆菌中表达。由于她的工程背景，奥伊斯基兴知道，在Chalfie实验室的显微镜还不够好，使用这种类型的实验，让她捕捉绿菌，从她的前工程实验室使用显微镜的图像。这项工作表明，GFP荧光要求没有绿色荧光蛋白本身以外的组件。事实上，其他实验室曾遇到的困难源于其使用限制性内切酶消化，亚克隆，这带来了一个额外的序列，阻止沿绿色荧光蛋白的荧光表达。奥伊斯基兴的在大肠杆菌中成功表达大肠杆菌 ，Chalfie的技术员元涂了表达绿色荧光蛋白在 C ， 线虫和Chalfie在1994年发表在"科学"的研究结果。 通过C 的研究线虫和GFP，Chalfie觉得有一个重要的教训，要对基础研究的重要性中汲取。虽然近期推临床相关或专利生产的（翻译）的研究，一直存在着，Chalfie警告，单独采取这种做法是错误的，如何"可悲的一点:"我们对生物学的了解。他指出，在生物学中未知的广袤，并指出，如GFP重要的发现是非常频繁使用的模式生物的多样化，通过基础研究。事实上，绿色荧光蛋白的生物发光的研究并没有原先对人体健康的直接应用。然而，我们对它的认识，导致广泛的临床相关发现和发展。 Chalfie认为我们不应该限制自己:"我们应该更自由一点，调查在不同方向的东西，和一点点敬畏，我们要找到什么。"

Watch this Scientific Journal Video about The 2009 Lindau Nobel Laureate Meeting: Martin Chalfie, Chemistry 2008 at JoVE.com

The 2009 Lindau Nobel Laureate Meeting: Martin Chalfie, Chemistry 2008

美国生物学家马丁Chalfie罗杰钱学森和下村修分享2008年诺贝尔化学奖的绿色荧光蛋白（GFP）的发现和发展。 Chalfie克隆的GFP编码序列，并表示它在这两个<em&gt; E。大肠杆菌</em&gt;<em&gt; C。线虫</em&gt;，第一次证明，没有其他的因素是GFP的发光。

2009年林岛诺贝尔奖得主会议:2008年，化学马丁Chalfie

American Biologist Martin Chalfie shared the 2008 Nobel Prize in Chemistry with Roger Tsien and Osamu Shimomura for their discovery and development of the ...

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6.7K 次观看. 美国生物学家马丁Chalfie罗杰钱学森和下村修分享2008年诺贝尔化学奖的绿色荧光蛋白（GFP）的发现和发展。 Chalfie克隆的GFP编码序列，并表示它在这两个 E。大肠杆菌 C。线虫，第一次证明，没有其他的因素是GFP的发光。

视频: The 2009 Lindau Nobel Laureate Meeting: Martin Chalfie, Chemistry 2008

December 2011: This Month in JoVE

The Journal of Visualized Experiments (JoVE) closes 2011 with a December issue that begins by honoring World's AIDS Day with an article that deals with visualizing the structure of HIV envelope glycoproteins, which are a critical component of the viral infection pathway.
One key method for determining virion envelope structure is cryo-electron tomography. Prior to using this technique, soluble virus must be vitrified on a gold grid - a process that involves adding gold to a viral suspension, transferring that suspension to the grid, and plunging the grid into liquid ethane. The virus particles, present in a thin film of liquid atop the grid, are frozen at a rate greater than 100,000 K per second. Following vitrification, collected samples are loaded into the electron microscope and precisely defined regions on the grid are hit with a focused electron beam, while the grid is tilted. The tilt series that results from these imaging sessions is comprised of hundreds of individual images, which must be aligned and averaged to resolve individual envelope proteins. This computationally intense process often requires computer clusters, capable of remote-access parallel processing, like NIH's Biowulf.
Once completed, 3D models can not only provide insight into the structure of envelope glycoproteins themselves, but can also reveal the manner in which they interact other proteins, like the neutralizing antibody B12, which is information that can greatly influence vaccine design.
This article, from the laboratory of Dr. Sriram Subramaniam involves contributions from twenty authors, including not only graduates students and postdocs, but also middle and highschool students, showing that you are never too young to make an impact in the fight against a disease that has infected over 60 million people, worldwide.
In Clinical and Translational Medicine, JoVE presents an article from the University College London and the University of Oxford that investigates the physiological correlates of nociception, or pain sensation, in infants. Typically, infants respond to noxious stimuli by crying and changing their facial expressions - reactions considered to be autonomic and reliant on subcortical regions of the brain.
In order to understand the involvement of the cerebral cortex and spinal cord in infant nociception our authors first prepare infants for EEG and EMG recordings. They also set up physiological monitoring and video recording, so that the infant's facial expressions can be linked to physiological data. Following assessment of infant responses to neutral tactile stimuli, the baby is subjected to heel lancing - a clinical procedure used to obtain blood samples from infants. After data processing, electrical potentials can be compared between noxious and non-noxious stimuli in these infants, which will hopefully lead to meaningful conclusions, regarding how nociceptive information is interpreted by newborns.
JoVE's Bioengineering section takes a more clinical focus in December with an article from the University of Wisconsin, Madison, which involves the fabrication of a compartmentalized microfluidic device for studying cancer stem cell migration. Following device design, master molds are created by photolithography for the application of poly dimethyl siloxane (PDMS)- an organosilicon compound that is transparent and biologically inert - making it suitable for microfluidics applications with live cells. Specifically, our authors load tumor stem cells derived from multiform glioblastoma, a highly malignant brain cancer, into the compartmentalized devices and then place them in a specialized imaging system that acquires data over a few days. 
Time lapse video from these authors show that cancer stem cells are able to regulate their morphology in the size-constrained microfluidic channels much like they would in the interstitial space, and fluidic isolation made possible by chambers like these facilitates drug screening for potential therapeutics that can affect cancer stem cell migration.
Late on in the month, JoVE travels to France, where investigators from the Institute Pasteur and the Centre National de la Recherche Scientifique, illustrate a method for inducing expression of channel-rhodopsin, a light-activated ion channel, into the olfactory bulb of mice, as well as subsequent neuronal stimulation using a miniature light emitting diode (LED).
Our authors give detailed step-by-step instructions for preparing injection needles, and for precise injection of virus into the rostral migratory stream, a site of migration for adult born neuroblasts. Furthermore, they show how miniature LED stimulation devices are fabricated, calibrated, and implanted atop a cranial window prepared over the bulb. In particular, our authors are interested in the way that adult born neurons integrate into existing olfactory neuronal circuits, and they show that LED stimulation in awake behaving animals, can lead to activation of virally-tranduced neuroblasts in the olfactory bulb, which have migrated from the rostral migratory stream.
These video-articles comprise JoVE's highlights for the month of December. Other noteworthy upcoming articles illustrate methods for patterning conductive inks into microelectrodes, measuring cytosolic calcium in contractile lymphatics, and imaging neuronal responses to pheromones in the vomeronasal organ.

Here are some highlights from the December 2011 Issue of Journal of Visualized Experiments (JoVE).

这在朱庇特12月

The Journal of Visualized Experiments (JoVE) closes 2011 with a December issue that begins by honoring World's AIDS Day with an article that deals with ...

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August 2012: This Month in JoVE

Traditional microscopy requires lens objectives to magnify specimens, and can involve numerous optical components like additional objectives, filters, and mirrors to refract and direct light to optical sensors. The August 2012 issue of JoVE (Journal of Visualized Experiments) is marked by the third publication from the Ozcan Lab (University of California, Los Angeles) on their lens free lens-free "on-chip" microscopy platform, which they have pioneered.
Back in 2009, the Ozcan lab published their first article, which demonstrated the use of a lens-free imaging platform that could image cells within whole blood. This imaging technique involves placing a sample directly on an optoelectric sensor array, like the CCD or CMOS chips found in the standard camcorder. Then, LED illumination from above the sample casts shadows on the chip, which are recorded as digital holograms that can be subjected to image processing. Not only does this lensless imaging system allow for an extremely large field of view (FOV of 0.6-8 cm2), the lack of bulky optical components makes it excellent for microfluidic applications. The Ozcan group demonstrates this in their follow-up 2011 JoVE article, in which fluorescently labeled white blood cells are imaged within a microfluidic chip.
This August in JoVE, the Ozcan group demonstrates further advancements to their lens-free microscopy platform by applying it to tomographic imaging of the nematode, C. elegans. By changing the angle of illumination, using a motorized rotation stage, the Ozcan group shows how multiple holographic slices may be generated for a sample-which, following processing, can be combined into tomograms. The Ozcan group confirms that their 3D reconstruction of the worm is successful by citing that the pharyngeal tube, a 5-micron-thick structure, is only visible in a single slice and in a 3D reconstructed cross section.
The compact and inexpensive nature of the lens-free imaging systems - designed and assembled by the Ozcan group - make them well suited for point-of-care diagnostic applications, or even cell phone based microscopy that can be performed in the field.
At the Ernest Gallo Clinic and Research Center at University of California, San Francisco JoVE learns how to visualize morphological changes in single cells using three-dimensional fluorescence microscopy and computer analysis. In this procedure, cells are given agonist, antagonist plus agonist, or no treatment, and prepared for immunofluorescence microscopy. Three-dimensional images are acquired with the help of a two-photon laser system, and the fluorescence data are processed on the computer using Imaris and Matlab.
To demonstrate the power of this technique, our authors quantify single-cell phenotypic changes in transfected HEK293 cells. Three-dimensional morphometric analysis shows dramatic changes in cells treated with agonist. These changes are not always apparent when using conventional 2D analysis. This technique allows morphological changes in single cells to be quantified and characterized in response to pharmacologic stimuli, offering a powerful new assessment tool to the arsenal of drug discovery research.
This month, in JoVE Applied Physics, we travel to South Carolina to visit the School of Materials Science and Engineering at Clemson University. Here, researchers show us how to construct lithium batteries for laboratory research.
The basic components of a lithium coin cell are the working electrode, the lithium electrolyte, and a lithium foil counter electrode. The different components of the coin cell are carefully assembled and sealed with a crimping machine. The finished coin cell can then be characterized in a battery tester. Using their homemade coin cells, our researchers can experiment with using different materials to make the cathodes of lithium coin cells.
This month in JoVE Immunology and Infection, we head to Basel, Switzerland to witness two techniques for diagnosing helminth infections. We also see how investigators perform the shuttle run test in children, in order to assess the effects of these infections on physical fitness.
Helminths are parasitic worms that can live inside humans and animals. Many species infect the intestinal tract, and may be transmitted through soil. Our authors demonstrate the Kato-Katz technique, which is used for diagnosing several common soil-transmitted helminths. They also demonstrate the Baermann technique, which uses light to stimulate the migration of certain types of phototactic helminths out of sample preparations. The various types of helminth species can subsequently be determined based on morphology.
Our authors then demonstrate how to set up and perform a 20-meter shuttle run test for children, and to measure the impact of helminth infections on physical fitness. This method can help determine the true burden of soil-transmitted helminth infections on overall health.
This brief summary highlights but a few notable video-articles that will be released in August. We'll also feature articles that demonstrate how to express genes in the midbrain of the chick embryo, study neutrophil transmigration in parallel flow chambers lined with endothelial cells, and precisely measure proteolytic degradation of the extracellular matrix.

Traditional microscopy requires lens objectives to magnify specimens, and can involve numerous optical components like additional objectives, filters, and mirrors to refract and direct light to optical sensors. The August 2012 issue of JoVE (Journal of Visualized Experiments) is marked by the third publication from the Ozcan Lab (University of California, Los Angeles) on their lens-free "on-chip" microscopy platform, which they have pioneered.

2012年8月:本月在朱庇特

Traditional microscopy requires lens objectives to magnify specimens, and can involve numerous optical components like additional objectives, filters, and ...

2012: A Year In Review

Here's a look at some of the milestones and highlights of the year 2012 in Journal of Visualized Experiments (JoVE).
At the start of 2012, JoVE reached a major milestone by publishing our 1500th article. In this publication, in JoVE Bioengineering, Yu et al. described a method for analyzing the free radical composition of cigarette smoke. This method simulated cigarette puffing with a single-port smoking device, and combined it with electron spin resonance (ESR) spectroscopy to measure the degree to which antioxidant compounds can scavenge free radicals in cigarette smoke. A possible outcome of this research is the creation of less harmful cigarettes.
JoVE Bioengineering also contained the most-viewed article of 2012. Owczarczak et al. described a method for hacking a standard inkjet printer, loading ink cartridges with a cell-suspension "bioink," and printing lines of live cells onto glass slides. The thermal inkjet printing process creates temporary pores in the cell membrane, visualized by the incorporation of fluorescently labeled actin monomers. This concept, called "bioprinting," has many potential applications in cell and tissue engineering.
Also in 2012, JoVE Bioengineering featured a protocol by Hsia et al. for isolating and purifying spider silk protein, spinning it into fibers, and assessing the fiber strength. This protocol for laboratory-scale production of spider silk, which is stronger than tensile steel, can potentially be extended to large-scale manufacturing.
In JoVE Clinical and Translational Medicine, we featured a number of articles related to stroke, one of which. In one article, M&ouml;bius-Winkler et al. demonstrated placement of the WATCHMAN left atrial appendage occlusion device from Boston Scientific. This device is designed to prevent strokes by trapping clots within the left atrial appendage of the heart before they exit. Our authors demonstrated how to insert the guide wire through the femoral artery into the heart, advance the device into the left atrium, and deploy the device in the left atrial appendage.
Another article focused on stroke, Hamel et al. described the use of a driving simulator to study how patients compensate for visual field defects following stroke. By analyzing the compensatory gaze behavior of patients as they navigated through virtual driving courses with varying degrees of complexity, our authors see great potential for the use of driving simulators in stroke rehabilitation.
In JoVE Immunology and Infection, we published an article by Keyel et al. describing the real-time kinetics of immune cell responses to bacterial toxins using live cell microscopy. Combined with high-speed 3D confocal microscopy, this technique can also visualize the cellular repair response.
Also in JoVE Immunology and Infection, we featured an article by Yap et al. describing methods for diagnosing helminth infections in children. Helminths are parasitic worms that can infect the intestinal tract of humans and animals, and may be distinguished based on their morphology. The same article demonstrated how to measure the impact of helminth infections on physical fitness in children, which is a measure of overall health.
In JoVE Neuroscience, we featured a method by Hoffmann et al., who used tiny headphones to alter the auditory feedback in songbirds, and analyzed the computational and neurophysiological basis of vocal learning in birds when they adjust their singing in response to altered acoustic signals.
Also in JoVE Neuroscience, we moved from birds to bees with a method that assesses associative and non-associative tactile learning in honeybees. This method, by Mujagi&#263; et al., allowed bees to scan different metal surfaces with their antennae, then conditioned them to expect sugar water when their antennae touched particular surfaces, and analyzed the corresponding changes in antennal movement.
In 2012, we launched a brand-new section: JoVE Applied Physics, which features articles on subjects ranging from plasma physics to material science. The Applied Physics section also contained JoVE's 2000th video article, filmed in a synchrotron radiation facility, where Borisenko et al. showed how to determine the electronic structure of complex materials using angle-resolved photoemission spectroscopy.
In the JoVE General section, we published an article by Artioli et al. demonstrating the use of a portable gas analyzer to measure oxygen consumption during complex exercise - such as judo. By analyzing oxygen consumption and collecting blood for measuring plasma lactate concentration, this method can determine the relative contributions of different energy systems to specific aspects of complex exercise.
Finally, JoVE followed Dolhi et al. to a permanently ice-covered saline lake in Antarctica. There, scientists drilled through the ice to study single-celled microorganisms called protists that live in the harsh conditions of Antarctica.
This Year in Review was just a brief glimpse of a few of over 600 video articles that JoVE offered in 2012. Browse the JoVE archives to see thousands of other videos, and stay tuned for what's coming up this year in JoVE: The Journal of Visualized Experiments.

Here's a look at some of the milestones and highlights of the year 2012 in Journal of Visualized Experiments (JoVE).

2012年:年度回顾

 
 
Here's a look at some of the milestones and highlights of the year 2012 in Journal of Visualized Experiments (JoVE).
At the start of 2012, JoVE ...

2009年林岛诺贝尔奖得主会议:2008年，化学马丁Chalfie

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