<h2 class="jove_title"><a href="https://www.jove.com/video/3122">Selection of Plasmodium falciparum Parasites for Cytoadhesion to Human Brain Endothelial Cells</a></h2>
Antoine Claessens, J. Alexandra Rowe Centre for Immunity, Infection and Evolution, University of Edinburgh
An in vitro model for cerebral malaria sequestration is described1. Plasmodium falciparum infected red blood cells are selected for binding to immortalized human brain microvascular endothelial cells. The selected parasites show a distinct phenotype. The selection process can be applied using various P. falciparum strains and endothelial cell lines.
<h2 class="jove_title"><a href="https://www.jove.com/video/3308">Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice</a></h2>
Michael T. Kuhlmann1, Simon Cuhlmann2, 3, Irmgard Hoppe1, Rob Krams3, Paul C. Evans2, Gustav J. Strijkers4, Klaas Nicolay4, Sven Hermann1, Michael Sch&#228;fers1 1European Institute for Molecular Imaging, Westf&#228;lische Wilhelms-University M&#252;nster, 2British Heart Foundation Cardiovascular Sciences Unit, Imperial College London , 3Department of Bioengineering, Imperial College London , 4Biomedical Engineering, Eindhoven University of Technology
The constricting cuff presented in this article is designed to induce atherosclerosis in the murine common carotid artery. Due to the conical shape of its inner lumen the implanted cuff generates well-defined regions of low, high and oscillatory shear stress triggering the development of atherosclerotic lesions of different inflammatory phenotypes.
<h2 class="jove_title"><a href="https://www.jove.com/video/3330">Analysis of Trunk Neural Crest Cell Migration using a Modified Zigmond Chamber Assay</a></h2>
Christopher C. Walheim1, Juan Pablo Zanin2, Maria Elena de Bellard1 1Department of Biology, California State University, Northridge, 2Centro de Biolog&#237;a Celular y Molecular, Universidad Nacional de C&#243;rdoba
An approach to analyze the migration of explanted cells (trunk neural crest cells) is described. This method is inexpensive, gentle, and capable of distinguishing chemotaxis from both chemokinesis and other influences on migratory polarity such as those derived from cell-cell interactions within the primary trunk neural crest cell culture.
<h2 class="jove_title"><a href="https://www.jove.com/video/3350">Organotypic Slice Cultures of Embryonic Ventral Midbrain: A System to Study Dopaminergic Neuronal Development in vitro</a></h2>
Gabriela Oana Bodea, Sandra Blaess Institute of Reconstructive Neurobiology, University of Bonn
A method to generate organotypic slices from the E12.5 murine embryonic midbrain is described. The organotypic slice cultures can be used to observe the behavior of dopaminergic neurons or other ventral midbrain neurons.
<h2 class="jove_title"><a href="https://www.jove.com/video/3362">Visualization of Caenorhabditis elegans Cuticle Structures using the Lipophilic Vital Dye, DiI</a></h2>
Robbie D. Schultz1, 2, Tina L. Gumienny2 1Department of Molecular and Cellular Medicine, Texas A&#38;M University System Health Science Center, 2Department of Molecular and Cellular Medicine, Texas A&#38;M University System Health Science Center, College of Medicine
We present a method to visualize cuticle in live C. elegans using the red fluorescent lipophilic dye DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate), which is commonly used in C. elegans to visualize environmentally exposed neurons. With this optimized protocol, alae and annular cuticular structures are stained by DiI and observed using compound microscopy.
<h2 class="jove_title"><a href="https://www.jove.com/video/3406">A Protocol for Detecting and Scavenging Gas-phase Free Radicals in Mainstream Cigarette Smoke</a></h2>
Long-Xi Yu1, Boris G. Dzikovski2, Jack H. Freed3, 2 1CDCF-AOX Lab, 2National Biomedical Center for Advanced ESR Technology (ACERT), Department of Chemistry and Chemical Biology, Cornell University, 3ACERT Research, Center Department of Chemistry and Chemical Biology, Baker Laboratory, Cornell University
Spin-trapping ESR spectroscopy was used to study the effect of plant antioxidants lycopene, pycnogenol and grape seed extract on scavenging gas-phase free radicals in cigarette smoke.
<h2 class="jove_title"><a href="https://www.jove.com/video/3417">How to Measure Cortical Folding from MR Images: A Step-by-step Tutorial to Compute Local Gyrification Index</a></h2>
Marie Schaer1, Meritxell Bach Cuadra2, 3, Nick Schmansky4, Bruce Fischl4, Jean-Philippe Thiran2, Stephan Eliez1 1Department of Psychiatry, University of Geneva School of Medicine, 2Signal Processing Laboratory, &#201;cole Polytechnique F&#233;d&#233;rale de Lausanne, 3Department of Radiology, University Hospital Center and University of Lausanne, 4Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital
Measuring gyrification (cortical folding) at any age represents a window into early brain development. Hence, we previously developed an algorithm to measure local gyrification at thousands of points over the hemisphere1. In this paper, we detail the computation of this local gyrification index.
<h2 class="jove_title"><a href="https://www.jove.com/video/3551">The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness</a></h2>
Hani H. Al-Nakhli1, Jerrold S. Petrofsky1, 2, Michael S. Laymon2, Lee S. Berk1 1Loma Linda University, 2Azusa Pacific University
The purpose of this investigation was to assess whether using an infra-red thermal camera is a valid tool for detecting and quantifying the muscle soreness after exercising.

january 2012: this month in jove

The Journal of Visualized Experiments (JoVE) begins 2012 by publishing its 1500th article, which focuses on a method for analyzing the free radical composition of cigarette smoke. Although tobacco tar is thought to be the most carcinogenic agent in cigarette smoke, free radicals are well known cancer-causing molecules, and the level of these carcinogens contained in cigarette smoke is not currently known. The method employed in this article combines the use of a single-port smoking device, which simulates cigarette puffing, with electron spin resonance (ESR) spectroscopy, a technique, which measures the spin energy emitted by the unpaired electrons in free radicals. One important modification of the single port smoking device used in this article, is the introduction of a liquid nitrogen trap in front of the spin trap. Liquid nitrogen removes water vapor from cigarette smoke and greatly improves ESR spectra as a result.
In addition to measuring the free radical concentration found in cigarette smoke, plant antioxidants are introduced into modified cigarette filters and the ESR spectra of cigarette smoke passed through these filters is obtained. Grape seed extract and lycopene were observed to have the most significant scavenging effects on free radicals, and if cost effective means of including these molecules into cigarette filters can be developed, a less harmful cigarette could be created.
In Clinical and Translational Medicine, JoVE presents an analytical method for determining the local Gyrification Index (lGI) - a measurement of the extent of cortical folding. Cortical folds develop during the first few post natal months, and adverse events during this time can lead to abnormalities in the brain's surface anatomy, which may ultimately correlate with neuropsychiatric disorders.
The analysis tool developed for calculating the local Gyrification Index is Freesurfer, which can convert T1 weighted MRI scans into 3D reconstructions of the cortex. Separate reconstructions are created for the grey/white mater and grey/pial interfaces, which are manually adjusted before computating the lGI. Following a smoothing operation, circular regions of the pial and grey/white surfaces are compared, in order to compute the lGI for thousands of points over the entire brain. Freesurfer makes analysis of gyrification straightforward and accessible to clinicians and brain scientists, who are interested in comparing the extent of cortical folding between different individuals in normal and disease states.
JoVE's Immunology and Infection article for the month of January continues the journal's tradition of publishing cutting edge methodology in vector born disease research. Specifically, this article adds to JoVE's comprehensive collection of protocols for malaria research by extending our library to cerebral malaria - a devastating condition with high susceptibility in children. Briefly, this protocol involves exposing a monolayer of human brain endothelial cells, passaged onto petri dishes, to Plasmodium falciparum-infected red blood cells. Following washing and enriching steps, high yields of parasites, which carry the potential to infect the brain, can be isolated and further characterized.
In Neuroscience, JoVE showcases a method for studying the migration of neural crest cells. These cells are a transient, multipotent, developmental cell type that can, in addition to becoming neuronal tissue, differentiate into smooth muscle, chromaffin cells, cartiliage, and bone. Chick embryos are carefully prepared, so that the trunk region can be isolated and the neural tube excised for overnight culture on fibronectin-coated coverslips, which allows neural crest cells to emigrate from the neural tube. Following removal of the neural tube, coverslips containing the cultured neural crest cells are transferred to a modified Zigmond chamber - a chamber which enables a concentration gradient of putative chemotactic agent to be established across the culture. Timelapes imaging followed by analysis of cell trajectories, is used to assess the chemotactic potential of molecules loaded into the zigmund chamber, providing insight into the molecular cues that guide migration of this important developmental cell type.
These featured articles comprise four of fifty video-protocols for release in January. Other notable articles include methods for visualizing the cuticle structures of c. elegans, thermal imaging of delayed onset muscle soreness, culturing the ventral midbrain in organotypic slices, and inducing atherosclerosis in mice.

Watch this Scientific Journal Video about January 2012: This Month in JoVE at JoVE.com

January 2012: This Month in JoVE

Questo gennaio a JOVE

The Journal of Visualized Experiments (JoVE) begins 2012 by publishing its 1500th article, which focuses on a method for analyzing the free radical ...

Research

JoVE Journal

Biology

Video: January 2012: 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 ...

Ricerca

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