<h2 class="jove_title"><a href="https://www-jove-com.remotexs.ntu.edu.sg/details.php?id=3110">Transfecting and Nucleofecting Human Induced Pluripotent Stem Cells</a></h2>
Papri Chatterjee, Yuri Cheung, Chee Liew UCR Stem Cell Center, Department of Cell Biology and Neuroscience, University of California Riverside
Despite recent advancements in genetic modification, transfection of human embryonic stem cells (HESCs) remains a capricious process. To our knowledge, systematic and efficient methods to transfect human induced pluripotent stem cells (iPSCs) have not been reported. Here, we describe robust protocols to efficiently transfect and nucleofect human iPSCs.
<h2 class="jove_title"><a href="https://www-jove-com.remotexs.ntu.edu.sg/details.php?id=2950">Microwave Assisted Rapid Diagnosis of Plant Virus Diseases by Transmission Electron Microscopy</a></h2>
Bernd Zechmann1, 2, Gerhard Graggaber1, G&uuml;nther Zellnig1 1Institute of Plant Sciences, University of Graz, 2Institute for Electron Microscopy and Fine Structure Research, Graz University of Technology
This study describes a method that allows the rapid and clear diagnosis of plant virus diseases in about half a day by using a combination of microwave assisted plant sample preparation for transmission electron microscopy and negative staining methods.
<h2><a href="https://www-jove-com.remotexs.ntu.edu.sg/details.php?id=3363">Following Cell-Fate in E. coli After Infection by Phage Lambda</a></h2>
Lanying Zeng1, Ido Golding1, 2, 3 1Department of Physics, University of Illinois at Urbana-Champaign, 2Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, 3Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine
This article describes the procedure for preparing a fluorescently-labeled version of bacteriophage lambda, infection of E. coli bacteria, following the infection outcome under the microscope, and analysis of infection results.
<h2 class="jove_title"><a href="https://www-jove-com.remotexs.ntu.edu.sg/details.php?id=3107">Stem Cell Transplantation Strategies for the Restoration of Cognitive Dysfunction Caused by Cranial Radiotherapy</a></h2>
Munjal M. Acharya*, Dante E. Roa*, Omar Bosch, Mary L. Lan*, Charles L. Limoli Department of Radiation Oncology, University of California Irvine * These authors contributed equally
Brain tumor patients routinely undergo cranial radiotherapy, and while beneficial, this treatment often results in debilitating cognitive dysfunction. This serious unresolved problem has at present, no clinical recourse, and has driven our efforts to devise stem cell based therapies for the recovery of radiation-induced cognitive decrements.
<h2 class="jove_title"><a href="https://www-jove-com.remotexs.ntu.edu.sg/details.php?id=3108">Quantifying Cognitive Decrements Caused by Cranial Radiotherapy</a></h2>
Lori- Ann Christie, Munjal M. Acharya, Charles L. Limoli Department of Radiation Oncology, University of California Irvine 
Cognitive impairment resulting from the radiotherapeutic management of brain tumors represents a clinically intractable condition that adversely impacts quality of life. The capability to critically evaluate potential interventions for ameliorating radiation-induced cognitive decrements ultimately depends on the capability to undertake rigorous quantitative assessments of cognition.
<h2 class="jove_title"><a href="https://www-jove-com.remotexs.ntu.edu.sg/details.php?id=3238">The Trier Social Stress Test Protocol for Inducing Psychological Stress</a></h2>
Melissa A. Birkett Department of Psychology, Northern Arizona University
This article describes a protocol for inducing psychological stress in participants, which enables researchers to measure psychological, physiological and neuroendocrine responses to stress within single participants or between groups.
<h2 class="jove_title"><a href="https://www-jove-com.remotexs.ntu.edu.sg/details.php?id=2967">Oral Biofilm Analysis of Palatal Expanders by Fluorescence In-Situ Hybridization and Confocal Laser Scanning Microscopy</a></h2>
Barbara Klug1, 2, Claudia Rodler1, Martin Koller3, Gernot Wimmer3, Harald H. Kessler2, Martin Grube4, Elisabeth Santigli1 1Department of Orthodontics and Maxillofacial Orthopedics, Medical University of Graz, 2Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, 3Department of Prosthodontics, Restorative Dentistry, Periodontology and Implantology, Medical University of Graz, 4Institute of Plant Sciences, Karl-Franzens-University Graz
We present a protocol for structural and compositional analysis of natural oral biofilm from orthodontic appliances with in situ hybridization (FISH) and confocal laser scanning microscopy (CLSM). Oral biofilm samples were collected from palatal expanders, scraping acrylic-resin flakes off their surface and referring them for molecular processing.
<h2 class="jove_title"><a href="https://www-jove-com.remotexs.ntu.edu.sg/details.php?id=2615">Quantitatively Measuring In situ Flows using a Self-Contained Underwater Velocimetry Apparatus (SCUVA)</a></h2>
Kakani Katija1, Sean P. Colin2, 3, John H. Costello3, 4, John O. Dabiri5 1Applied Ocean Physics and Engineering, Woods Hole Oceanographic Institution, 2Environmental Science and Marine Biology, Roger Williams University, 3Marine Biology Laboratory, Whitman Center, 4Department of Biology, Providence College, 5Departments of Aeronautics and Bioengineering, California Institute of Technology
This protocol provides instructions on how to use a self-contained underwater velocimetry apparatus (SCUVA), which is designed for quantification of in situ animal-generated flows. In addition, this protocol addresses challenges posed by field conditions, and includes operator motion, predicting position of animals, and orientation of SCUVA.
<h2 class="jove_title"><a href="https://www-jove-com.remotexs.ntu.edu.sg/details.php?id=3336">Imaging Odor-Evoked Activities in the Mouse Olfactory Bulb using Optical Reflectance and Autofluorescence Signals</a></h2>
Romain Chery, Barbara L&#x2019;Heureux, Mounir Bendahmane, R&#233;mi Renaud, Claire Martin, Fr&#233;d&#233;ric Pain, Hirac Gurden Laboratoire d&#x2019;Imagerie et de Mod&#233;lisation en Neurobiologie et Canc&#233;rologie, UMR8165 Universit&#233; Paris Sud 11, Paris Diderot 7 &#x2013; CNRS
This article presents the protocols of intrinsic optical signals and flavoproteins autofluorescence signals imaging to map odor-evoked activities at the surface of the olfactory bulb in mice.

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october 2011: this month in jove

The October 2011 issue of the Journal of Visualized Experiments (JoVE) presents the journal's first effort to explore oceanographic physics and engineering. Investigators at the Woods Hole Oceanographic Institute demonstrate the use of a self-contained underwater velocimetry apparatus, or SCUVA. Principle components of this instrument include an HD camera and laser, which when aligned with an optical lens, produces a vertical sheet of light. Following alignment and testing for a watertight seal, these researchers seek out inhabitants the water column for analysis. Nightly experimentation is necessary, so that natural light does not interfere with the laser sheet. SCUVA is specifically designed to capture the movement of particles in the water, so that the velocity fields - a vector field that describes the movement of fluid - can be determined around an organism in their natural environment.
In Clinical and Translational Medicine, orthodontic researchers from the University of Graz determine the composition of the oral biofilms - a surface-adhering aggregate of microorganisms - that form on their patient's palatal expanders. These authors use fluorescent in situ hybridization combined with confocal microscopy to detect specific bacteria within the biofilm as well as visualize its 3-dimensional structure.
JoVE also visits Graz for Immunology and Infection, where researchers at the University of Graz and the Graz institute of technology use transmission electron microscopy, or TEM, to analyze the ultrastructural changes that take place in the plant leaf as a result of viral infection. Small sections of leaves are cut from the plant and subjected to a microwave-assisted fixation procedure, which drastically reduces sample preparation time for TEM. Following embedding and sectioning, samples are loaded into the electron microscope, and imaged. Aided by image analysis software, these authors can calculate the dimensions of virus particles as well as assess the ultrastructural changes in the plant leaf, which is critical for identifying the infecting virus.
In Neuroscience, researchers from the University of California, Irvine, present a series of behavioral paradigms aimed to measure the degree of cognitive impairment in rodents treated with cranial radiotherapy. These authors demonstrate a novel place recognition task (for spatial recognition memory), an elevated plus maze task (for measurement of anxiety), a fear-conditioning task (learning and memory), and the Morris Water Maze (for spatial learning). The cognitive deficits in rodents resemble those manifested in cancers survivors following cranial radiotherapy, and therefore standardization of such cognitive behavioral paradigms is critical to assess therapy. One potential therapy is stem cell transplantation into the hippocampus, which the authors demonstrate in a separate video-article in October.
On the topic of stem cells, researchers from the University of California, Riverside demonstrate methods for genetic manipulation of human induced pluripotent stem cells. Induced pluoripotent stem cells, IPSCs, can be derived from somatic cells, like the fibroblasts you see here, following forced expression of certain transcription factors, which cause the cells to be transformed into a pluripotent state. These authors show how iPSCs can be cultured in feeder-free conditions and provide helpful hints for lifting and seeding the cells at concentrations optimal for transfection and nucleofection
In addition to these featured publications, JoVE presents methods for taking physiological measurements in individuals under psychosocial stress, tracking bacterial lysis following infections with phage, imaging in the olfactory bulb via a cranial window, and dozens more video-articles of complex methods.

Watch this Scientific Journal Video about October 2011: This Month in JoVE at JoVE.com

October 2011: This Month in JoVE

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The October 2011 issue of the Journal of Visualized Experiments (JoVE) presents the journal's first effort to explore oceanographic physics and ...

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Video: October 2011: This Month in JoVE

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.

Agosto 2012: Questo mese in JOVE

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

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Biologia

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: 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 ...

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Agosto 2012: Questo mese in JOVE

2012: A Year in Review