<h2 class="jove_title"><a href="https://www.jove.com/video/51888">Generation of Transgenic Hydra by Embryo Microinjection</a></h2>
Celina E. Juliano1, Haifan Lin1, Robert E. Steele2
1Yale Stem Cell Center and Department of Cell Biology, Yale University School of Medicine, 2Department of Biological Chemistry and the Developmental Biology Center, University of California, Irvine
Stably transgenic Hydra are made by microinjection of plasmid DNA into embryos followed by random genomic integration and asexual propagation to establish a uniform line. Transgenic Hydra are used to track cell movements, overexpress genes, study promoter function, or knock down gene expression using RNAi.
<h2 class="jove_title"><a href="https://www.jove.com/video/51892">A Multi-Modal Approach to Assessing Recovery in Youth Athletes Following Concussion</a></h2>
Nick Reed1,2,3, James Murphy1, Talia Dick1, Katie Mah3, Melissa Paniccia3, Lee Verweel3, Danielle Dobney3, Michelle Keightley1,2,3
1Bloorview Research Institute, Holland Bloorview Kids Rehabilitation Hospital, 2Department of Occupational Science and Occupational Therapy, University of Toronto, 3Graduate Department of Rehabilitation Science, University of Toronto
This article provides an overview of a multi-modal approach to assessing recovery following concussion in youth athletes. The described protocol uses pre- and post-concussion assessment of performance across a wide variety of domains and can inform the development of improved concussion rehabilitation protocols specific to the youth sport community.
<h2 class="jove_title"><a href="https://www.jove.com/video/51826">Combination of Microstereolithography and Electrospinning to Produce Membranes Equipped with Niches for Corneal Regeneration</a></h2>
&#205;lida Ortega1, Farshid Sefat1, Pallavi Deshpande1, Thomas Paterson1, Charanya Ramachandran3, Anthony J. Ryan2, Sheila MacNeil1, Frederik Claeyssens1
1Department of Materials Science and Engineering, University of Sheffield, 2Department of Chemistry, University of Sheffield, 3L. V. Prasad Eye Institute
We report a technique for the fabrication of micropockets within electrospun membranes in which to study cell behavior. Specifically, we describe a combination of microstereolithography and electrospinning for the production of PLGA (Poly(lactide-co-glycolide)) corneal biomaterial devices equipped with microfeatures.
<h2 class="jove_title"><a href="https://www.jove.com/video/51824">Multimodal Optical Microscopy Methods Reveal Polyp Tissue Morphology and Structure in Caribbean Reef Building Corals</a></h2>
Mayandi Sivaguru1, Glenn A. Fried1, Carly A. H. Miller1,2, Bruce W. Fouke1,2,3
1Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 2Department of Geology, University of Illinois at Urbana-Champaign, 3Department of Microbiology, University of Illinois at Urbana-Champaign
An integrated suite of imaging techniques has been applied to determine polyp morphology and tissue structure in the Caribbean corals Montastraeaannularis and M. faveolata. Fluorescence, serial block face, and two-photon confocal laser scanning microscopy have identified lobate structure, polyp walls, and estimated chromatophore and zooxanthellae densities and distributions.

september 2014: this month in jove - visualizing coral, injecting hydra, engineering corneal implants, and assessing traumatic brain injury in children

Here's a look at what's coming up in the <a href="http://www.jove.com/archive/91">September 2014 issue</a> of <a href="http://www.jove.com/">JoVE: The Journal of Visualized Experiments</a>.
As summer comes to an end in the Earth's northern hemisphere, <a href="http://www.jove.com/enviro">JoVE Environment</a> takes us to the southern Caribbean Sea where year-round warm temperatures support the growth of corals, which are marine invertebrates in the phylum Cnidaria. Although coral reefs occupy less than 0.1% of the ocean floor, they are home to about 25% of all marine life. These delicate ecosystems provide a wealth of natural resources but they are vulnerable to environmental threats, including climate change. To better predict how environmental factors will affect coral reefs, <a href="http://www.jove.com/video/51824">Sivaguru et al.</a> have developed a way to construct 3D images of coral with fine details of the coral structure. This method can help establish a baseline for healthy coral and to visualize changes caused by different environmental conditions.
In <a href="http://www.jove.com/biology">JoVE Biology</a> we examine Hydra, which are freshwater relatives of coral and also in the phylum Cnidaria Named after the multi-headed serpent that was killed by Hercules in Greek mythology, Hydra have been the subject of tissue regeneration studies since the 1700s. But it wasn't until 2006 that transgenic manipulation techniques were developed for Hydra. <a href="http://www.jove.com/video/51888">Juliano et al.</a> demonstrate how to microinject Hydra embryos with plasmid DNA and to generate stable transgenic lines Transgenic Hydra can then be used in studies of cell fate, gene expression, and tissue regeneration.
In <a href="http://www.jove.com/bioengineering">JoVE Bioengineering</a>, we examine methods for regenerating tissues of the cornea, the clear front window of the eye. The edge of the cornea, or the limbus, is critical for corneal health. It prevents the ingrowth of blood vessels, thus keeping the cornea clear, and it is the source of stem cells that renew the outer corneal epithelium. If the corneal limbus is damaged or diseased, blindness can result, so in this issue, <a href="http://www.jove.com/video/51826">Ortega et al.</a> focus on methods for bioengineering the corneal limbus. Using microstereolithography and electrospinning, they create membranes that mimic the limbus. When these membranes are placed on corneal wound models. A new multi-layered corneal epithelium forms. This may lead to therapeutic devices that can regenerate the corneal epithelium and preserve vision in conditions that affect the limbus.
In <a href="http://www.jove.com/ctm">JoVE Clinical &amp; Translational Medicine</a>, we feature a protocol on concussions, which may have long-term physical and psychological effects. Although concussions are very common sports-related injuries in children, there is surprisingly little known about how a young brain responds to a concussion. Therefore, <a href="http://www.jove.com/video/51892">Reed et al.</a> have developed methods for assessing cognitive performance, balance and various agility/motor skills This may help optimize the treatment and rehabilitation of pediatric concussions.
You've just had a sneak peek of the <a href="http://www.jove.com/archive/91">September 2014 issue</a> of JoVE. Visit the website to see the full-length articles, plus many more.

Watch this Scientific Journal Video about September 2014: This Month in JoVE - Visualizing Coral, Injecting Hydra, Engineering Corneal Implants, and Assessing Traumatic Brain injury in Children at JoV

September 2014: This Month in JoVE - Visualizing Coral, Injecting Hydra, Engineering Corneal Implants, and Assessing Traumatic Brain injury in Children

Here's a look at what's coming up in the <a href="http://www.jove.com/archive/91">September 2014</a> issue of JoVE.

Here's a look at what's coming up in the September 2014 issue of JoVE: The Journal of Visualized Experiments.
As summer comes to an end in the Earth's ...

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ビデオ: September 2014: This Month in JoVE - Visualizing Coral, Injecting Hydra, Engineering Corneal Implants, and Assessing Traumatic Brain injury in Children

assessing dominant-submissive behavior in adult rats following traumatic brain injury

Assessing Dominant-Submissive Behavior in Adult Rats Following Traumatic Brain Injury

The present protocol describes a rat model of fluid percussion-induced traumatic brain injury followed by a series of behavioral tests to understand the development of dominant and submissive behavior. Using this model of traumatic brain injury in conjunction with specific behavioral tests enables the study of social impairments following brain injury.

assessing changes in synaptic plasticity using an awake closed-head injury model of mild traumatic brain injury

Assessing Changes in Synaptic Plasticity Using an Awake Closed-Head Injury Model of Mild Traumatic Brain Injury

Here, it is demonstrated how an awake closed-head injury model can be used for examining the effects of repeated mild traumatic brain injury (r-mTBI) on synaptic plasticity in the hippocampus. The model replicates important features of r-mTBI in patients and is used in conjunction with in vitro...

a metric test for assessing spatial working memory in adult rats following traumatic brain injury

A Metric Test for Assessing Spatial Working Memory in Adult Rats Following Traumatic Brain Injury

Traumatic brain injury (TBI) is commonly associated with memory impairment. Here, we present a protocol to assess spatial working memory after TBI via a metric task. A metric test is a useful tool to study spatial working memory impairment after...

detecting behavioral deficits in rats after traumatic brain injury

Detecting Behavioral Deficits in Rats After Traumatic Brain Injury

The goal of the behavioral tests presented here is to detect functional deficits in rats after traumatic brain injury. Four specific tests are presented that detect deficits in behaviors to reflect the damage to specific brain areas at times extending to one year after...

investigating alterations in caecum microbiota after traumatic brain injury in mice

Investigating Alterations in Caecum Microbiota After Traumatic Brain Injury in Mice

Presented here is a protocol to induce diffuse traumatic brain injury using a lateral fluid percussion device followed by the collection of the caecum content for gut microbiome...

inducing post-traumatic epilepsy in a mouse model of repetitive diffuse traumatic brain injury

Inducing Post-Traumatic Epilepsy in a Mouse Model of Repetitive Diffuse Traumatic Brain Injury

This systematic protocol describes a new animal model of post-traumatic epilepsy after repetitive mild traumatic brain injury. The first part details steps for traumatic brain injury induction using a modified weight drop model. The second part provides instructions on the surgical approach for single- and multi-channel electroencephalographic data acquisition...

a novel in vitro model of blast traumatic brain injury

A Novel In Vitro Model of Blast Traumatic Brain Injury

This paper describes a novel model of primary blast traumatic brain injury. A compressed-air driven shock tube is used to expose in vitro mouse hippocampal slice cultures to a single shock wave. This is a simple and rapid protocol generating a reproducible brain tissue injury with a high throughput.

A Novel In Vitro Model of Blast Traumatic Brain Injury

assessing spatial memory impairment in a mouse model of traumatic brain injury using a radial water tread maze

Assessing Spatial Memory Impairment in a Mouse Model of Traumatic Brain Injury Using a Radial Water Tread Maze

Here we present a protocol for a mouse-specific test of cognition that does not require swimming. This test can be used to successfully distinguish controlled cortical impact-induced traumatic brain injury mice from sham...

lateral fluid percussion: model of traumatic brain injury in mice

Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice

Lateral fluid percussion (LFP), an established model of traumatic brain injury in mice, is demonstrated. LFP fulfills three major criteria for animal models: validity, reliability and clinical relevance. The procedure, consisting of surgical craniotomy, fixation of hub followed by induction of injury, resulting in focal and diffuse injuries, is...

a mouse model of single and repetitive mild traumatic brain injury

A Mouse Model of Single and Repetitive Mild Traumatic Brain Injury

Athletes absorb several hundred mild traumatic brain injuries (mTBI)/concussions every year; however, the consequence of these on the brain is poorly understood. Therefore, an animal model of single and repetitive mTBI that consistently replicates clinically relevant symptoms provides the means to advance the study of mTBI and...

electromagnetic controlled closed-head model of mild traumatic brain injury in mice

Electromagnetic Controlled Closed-Head Model of Mild Traumatic Brain Injury in Mice

The protocol describes mild traumatic brain injury in a mouse model. In particular, a step-by-step protocol to induce a mild midline closed head injury and the characterization of the animal model is fully...

a method to inflict closed head traumatic brain injury in drosophila

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

Here we describe a method to inflict closed head traumatic brain injury (TBI) in Drosophila. This method provides a gateway to investigate the cellular and molecular mechanisms that underlie TBI pathologies using the vast array of experimental tools and techniques available for...

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

systems analysis of the neuroinflammatory and hemodynamic response to traumatic brain injury

Systems Analysis of the Neuroinflammatory and Hemodynamic Response to Traumatic Brain Injury

This protocol presents methods to characterize the neuroinflammatory and hemodynamic response to mild traumatic brain injury and to integrate these data as part of a multivariate systems analysis using partial least squares...

stretch in brain microvascular endothelial cells (cend) as an in vitro traumatic brain injury model of the blood brain barrier

Stretch in Brain Microvascular Endothelial Cells cEND as an In Vitro Traumatic Brain Injury Model of the Blood Brain Barrier

In vitro traumatic brain injury models are being developed to reproduce in vivo brain deformation. Stretch-induced injury has been employed for astrocytes, neurons, glial cells, aortic, and brain endothelial cells. However, our system uses a blood brain barrier (BBB) model that possesses properties constituting a legitimate model of the BBB to establish an in vitro&#160;TBI...

Stretch in Brain Microvascular Endothelial Cells (cEND) as an In Vitro Traumatic Brain Injury Model of the Blood Brain Barrier

modified mouse model of repetitive mild traumatic brain injury incorporating thinned-skull window and fluid percussion

Author Spotlight: Optimizing Rodent Models for Investigating the Mechanisms and Rehabilitation in Repeated Mild Traumatic Brain Injury

This protocol presents a modified mouse model of repetitive mild traumatic brain injury (rmTBI) induced via a closed-head injury (CHI) method. The approach features a thinned-skull window and fluid percussion to reduce the inflammation commonly caused by meninges exposure, along with improved reproducibility and accuracy in modeling rmTBI in...

rmTBI Induction in Mice via a Closed-Head Injury Method

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evaluation of blood-brain barrier breakdown in a mouse model of mild traumatic brain injury

Author Spotlight: Advancing Traumatic Brain Injury Research by Mapping Blood-Brain Barrier Breakdown Using Dual-Dye Fluorescence

A method was developed to visualize dye extravasation due to blood-brain barrier (BBB) breakdown by administering two fluorescent dyes to mice at different time points. The use of glycerol as a cryoprotectant facilitated immunohistochemistry on the same...

Evaluation of Blood-Brain Barrier Breakdown in a Mouse Model of Mild Traumatic Brain Injury

traumatic peripheral nerve injury in mice

Traumatic Peripheral Nerve Injury in Mice

The present protocol describes traumatic peripheral nerve injuries (TPNIs), including precisely calibrated crush, strictly aligned and misaligned laceration, as well as grafted and non-grafted gaps of the sciatic nerve in mice. Custom-designed sensors are developed to gauge nerve trauma, induced with commonly available tools to ensure reproducible post-TPNI outcomes.

advanced diffusion imaging in the hippocampus of rats with mild traumatic brain injury

Advanced Diffusion Imaging in The Hippocampus of Rats with Mild Traumatic Brain Injury

The overall goal of this procedure is to obtain quantitative microstructural information of the hippocampus in a rat with mild traumatic brain injury. This is done using an advanced diffusion-weighted magnetic resonance imaging protocol and region-of-interest based analysis of parametric diffusion...

controlled cortical impact model for traumatic brain injury

Controlled Cortical Impact Model for Traumatic Brain Injury

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be...

corneal and limbal alkali injury induction using a punch-trephine technique in a mouse model

Author Spotlight: Advancing Research in Corneal Opacity Treatment and Regeneration 

This protocol describes a method to induce an accurate and reproducible corneal and limbal alkali injury in a mouse model. The protocol is advantageous as it allows for an evenly distributed injury to the highly curved mouse cornea and...

A Modified Biopsy Punch Method for Precise Alkali Burns in Mice

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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.

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

Research

JoVE Journal

Biology

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).

 
 
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 ...