Name: october 2014: this month in jove - visualizing infectious disease, transfecting with the gene gun, and a novel bioreactor system
Uploaded: 2014-10-01T16:00:00.000+00:00
Duration: 3 min 54 s
Description: Watch this Scientific Journal Video about October 2014: This Month in JoVE - Visualizing Infectious Disease, Transfecting with the Gene Gun, and a Novel Bioreactor System at JoVE.com

<h2 class="jove_title"><a href="https://www-jove-com.remotexs.ntu.edu.sg/video/52008">Cultivation of Mammalian Cells Using a Single-use Pneumatic Bioreactor System</a></h2>
Kristina M. Obom1, Patrick J. Cummings1, Janelle A. Ciafardoni1, Yasunori Hashimura2, Daniel Giroux2
1Center for Biotechnology Education, Johns Hopkins University, 2PBS Biotech, Inc.
Using a pneumatic bioreactor, we demonstrate the assembly, operation, and performance of this single-use bioreactor system for the growth of mammalian cells. 
<h2 class="jove_title"><a href="https://www-jove-com.remotexs.ntu.edu.sg/video/52084">Visualizing Non-lytic Exocytosis of Cryptococcus neoformans from Macrophages Using Digital Light Microscopy</a></h2>
Sabriya Stukes, Arturo Casadevall
Department of Microbiology and Immunology, Albert Einstein College of Medicine
We describe how to visualize macrophage-C. neoformans (Cn) interactions in real time, with specific emphasis on the process of non-lytic exocytosis using digital light microscopy. Using this technique individually infected macrophages can be studied to ascertain various aspects of this phenomenon.
<h2 class="jove_title"><a href="https://www-jove-com.remotexs.ntu.edu.sg/video/51714">Averaging of Viral Envelope Glycoprotein Spikes from Electron Cryotomography Reconstructions using Jsubtomo</a></h2>
Juha T. Huiskonen, Marie-Laure Parsy, Sai Li, David Bitto, Max Renner, Thomas A. Bowden
Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford
An approach is presented for determining structures of viral membrane glycoprotein complexes using a combination of electron cryo-tomography and sub-tomogram averaging with the computational package Jsubtomo.
<h2 class="jove_title"><a href="https://www-jove-com.remotexs.ntu.edu.sg/video/52148"> Regioselective Biolistic Targeting in Organotypic Brain Slices Using a Modified Gene Gun</a></h2>
Jason Arsenault1,2, Andras Nagy1, Jeffrey T. Henderson1, John A. O'Brien2
1Leslie Dan Faculty of Pharmacy, University of Toronto, 2MRC-Laboratory of Molecular Biology, Cambridge, UK
Recent improvements in organotypic brain slice preparations have permitted their exploitation for biotechnological applications. Organotypic slices maintain local structural characteristics of in vivo biology, including functional synaptic connections. Here we present a regioselective biolistic delivery method to label and genetically manipulate these slices.

october 2014: this month in jove - visualizing infectious disease, transfecting with the gene gun, and a novel bioreactor system

Here's a look at what's coming up in the <a href="https://www-jove-com.remotexs.ntu.edu.sg/archive/92">October 2014 issue</a> of <a href="https://www-jove-com.remotexs.ntu.edu.sg/">JoVE: The Journal of Visualized Experiments</a>.
Right now, the world is witnessing the worst Ebola virus outbreak in history. So this month in <a href="https://www-jove-com.remotexs.ntu.edu.sg/i2">JoVE Immunology &#38; Infection</a>, we highlight a technique than can help us better understand how viruses establish infection. Ebola and many other enveloped viruses, including HIV, use glycoprotein spikes on their surfaces to penetrate host cells. <a href="https://www-jove-com.remotexs.ntu.edu.sg/video/51714">Huiskonen et al.</a> use a computational approach to analyze viral envelope spikes, revealing their precise, 3D structure, which is critical for understanding the molecular interactions between certain viruses and their hosts. And this knowledge can guide the design of antiviral drugs and vaccines. Our authors demonstrate the technique with a virus from the family Bunyaviridae, but it can be applied to many other viruses that pose biological threats.
Viruses aren't the only pathogens we worry about in <a href="https://www-jove-com.remotexs.ntu.edu.sg/i2">JoVE Immunology &#38; Infection</a>. Bacteria and fungi can be just as contagious and deadly. Every year the fungus Cryptococcus neoformans infects more than 1 million people worldwide and causes over 600,000 deaths mostly in sub-Saharan Africa, where it is one of the leading causes of death in people with AIDS. Cryptococcal cells are generally killed by macrophage cells of the immune system; but some can survive latently in macrophages, then later release into the surroundings. This phenomenon is poorly understood, so <a href="https://www-jove-com.remotexs.ntu.edu.sg/video/52084">Stukes and Casadevall</a> have developed a way to infect macrophages with Cryptococcus in vitro and observe them over an extended period using time-lapsed microscopy. This is a promising technique for studying the complex interactions between Cryptococcus and host macrophages, and can be applied to other fungal pathogens as well.
In <a href="https://www-jove-com.remotexs.ntu.edu.sg/neuro">JoVE Neuroscience</a>, we feature a technique that combines ballistics, or the science of shooting, with genetic engineering. Bioballistic gene guns were originally developed to inject genetic information into plant cells. <a href="https://www-jove-com.remotexs.ntu.edu.sg/video/52148">Arsenault et al.</a> have applied this technique to mammalian tissues. Where terminally differentiated cells, like neurons, can difficult to transfect using conventional methods. By using an improved and patented bioballistic delivery method, our authors quickly and efficiently transfect various genes into distinct tissue regions.
A common goal in <a href="https://www-jove-com.remotexs.ntu.edu.sg/bioengineering">JoVE Bioengineering</a> is to culture bacteria or eukaryotic cells in large scales for industrial or biomedical applications. Some cultures, like bacteria and yeasts, can be grown in suspension, But many mammalian cells grow in aggregates, while others grow anchored to a substrate <a href="https://www-jove-com.remotexs.ntu.edu.sg/video/52008">Obom et al.</a> present a novel bioreactor system that can support all three types of cultures. They focus on anchorage-dependent cells, which have culture parameters that can be difficult to control, especially in large scales. Small beads called microcarriers, are added to the culture, to increase the surface area for cell growth, allowing anchorage-dependent cells to be grown in suspension. This system has tremendous potential for growing a variety of cell types and products for many applications.
You've just had a sneak peek of the <a href="https://www-jove-com.remotexs.ntu.edu.sg/archive/92">October 2014 issue</a> of JoVE. Visit the website to see the full-length articles, plus many more, in <a href="https://www-jove-com.remotexs.ntu.edu.sg/">JoVE: The Journal of Visualized Experiments</a>.

Watch this Scientific Journal Video about October 2014: This Month in JoVE - Visualizing Infectious Disease, Transfecting with the Gene Gun, and a Novel Bioreactor System at JoVE.com

October 2014: This Month in JoVE - Visualizing Infectious Disease, Transfecting with the Gene Gun, and a Novel Bioreactor System

Here's a look at what's coming up in the <a href="https://www-jove-com.remotexs.ntu.edu.sg/archive/92">October 2014</a> issue of JoVE.

Here's a look at what's coming up in the October 2014 issue of JoVE: The Journal of Visualized Experiments.
Right now, the world is witnessing the worst ...

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1.9K Views.  Massachusetts Eye and Ear. Here's a look at what's coming up in the October 2014 issue of JoVE.

Video: October 2014: This Month in JoVE - Visualizing Infectious Disease, Transfecting with the Gene Gun, and a Novel Bioreactor System

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.

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

October 2014: This Month in JoVE - Visualizing Infectious Disease, Transfecting with the Gene Gun, and a Novel Bioreactor System

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