<h2 class="jove_title"><a href="https://www.jove.com/video/51234">Transient Gene Expression in Tobacco using Gibson Assembly and the Gene Gun</a></h2>
Matthew d. Mattozzi1,2, Mathias J. Voges1,2,3, Pamela A. Silver1,2, Jeffrey C. Way1,2
1Synthetic Biology Platform, Wyss Institute for Biologically Inspired Engineering, Harvard University, 2Department of Systems Biology, Harvard Medical School, 3Department of Biotechnology, Delft University of Technology
This work describes a novel method for selectively targeting subcellular organelles in plants, assayed using the BioRad Gene Gun.
<h2 class="jove_title"><a href="https://www.jove.com/video/51597">Surgical Retrieval, Isolation and in vitro Expansion of Human Anterior Cruciate Ligament-derived Cells for Tissue Engineering Applications</a></h2>
Ashim Gupta1, Kevin Sharif2, Megan Walters2, Mia D. Woods1, Anish Potty2, Benjamin J. Main4, Saadiq F. El-Amin III1,2,3
1Department of Medical Microbiology, Immunology &amp; Cell Biology, Southern Illinois University School of Medicine, 2Division of Orthopaedics and Rehabilitation, Department of Surgery, Southern Illinois University School of Medicine, 3Department of Electrical and Computer Engineering, Biomedical Engineering Program, Southern Illinois University Carbondale, 4University of Illinois at Springfield
For future applications as a patch to repair partial tears of the Anterior Cruciate Ligament (ACL), human ACL derived cells were isolated from tissue obtained during reconstructive procedures, expanded in vitro and grown on tissue engineered scaffolds. Cellular adhesion and morphology was then performed to confirm biocompatibility on scaffold surface.&#160;&#160;&#160;
<h2 class="jove_title"><a href="https://www.jove.com/video/51686">Exfoliation of Egyptian Blue and Han Blue, Two Alkali Earth Copper Silicate-based Pigments</a></h2>
Darrah Johnson-McDaniel, Tina T. Salguero
Department of Chemistry, The University of Georgia
The preparation and exfoliation of CaCuSi4O10&#160;and BaCuSi4O10 are described. Upon stirring in hot water, CaCuSi4O10 spontaneously exfoliates into monolayers, whereas BaCuSi4O10&#160;requires ultrasonication in organic solvents. NIR imaging illustrates the NIR emission properties of these materials, and aqueous dispersions of these nanomaterials are useful for solution processing.
<h2 class="jove_title"><a href="https://www.jove.com/video/51337">Extracellular Wire Tetrode Recording in Brain of Freely Walking Insects</a></h2>
Peiyuan Guo, Alan J. Pollack, Adrienn G. Varga, Joshua P. Martin, Roy E. Ritzmann
Department of Biology, Case Western Reserve University
We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allows us to monitor activity in individual units of tethered cockroaches. Here we present a modified version of that technique that allows us to also record brain activity in freely moving insects.

april 2014: this month in jove - bioengineering for acl tears, neuroscience behind insect locomotion, improved genetic engineering of crops, and synthesizing secret ink

Here's a look at what's coming up in the <a href="http://www.jove.com/archive/86">April 2014 issue</a> of <a href="http://www.jove.com/">JoVE: The Journal of Visualized Experiments</a>.
In the <a href="http://www.jove.com/bioengineering">Bioengineering</a> section we have a video article that examines the anterior cruciate ligament (ACL), which is critical for keeping the knee stable when pivoting or planting. Torn or ruptured ACLs are among the most common of all serious knee injuries, especially in athletes, and even partial tears can hurt knee stability and function. Because they can't heal on their own, torn ACLs are often treated surgically, sometimes by reconstructing the ligament with a tendon grafted from the patient's own body or from a cadaver. Every year, over 100,000 ACL reconstructions are performed in the United States. The torn ligaments are usually discarded as surgical waste, but <a href="http://www.jove.com/video/51597/">Gupta et al.</a> see them as a valuable source of cells for potentially repairing ACLs. They developed a method for harvesting and expanding human ACL-derived cells then growing them on scaffolds to form tissue-engineered patches. Under scanning electron microscopy, the engineered matrix demonstrates good cellular adherence and growth. This technique could lead to methods for strengthening damaged ligaments and enhancing ACL repair, especially for those wishing to maintain an athletic lifestyle.
In the <a href="http://www.jove.com/neuro">Neuroscience</a> section we examine motor control in insects, which are among the most athletic creatures on Earth. They are especially good at adapting to sudden changes in terrain, so scientists are interested in correlating neural activity with behavior in insects. <a href="http://www.jove.com/video/51337/">Guo et al.</a> have developed a method for recording brain activity in the discoid cockroach, also known as the tropical cockroach, as they explore an arena or as they encounter obstacles that cause them to change course. The authors also demonstrate how to collect the data for subsequent analysis, which may yield insight into the neural control of behavior.
In <a href="http://www.jove.com/enviro">JoVE Environment</a> we feature a technique for transient gene expression in plants. To express gene products in specific parts of the cell, plants often have several copies of a gene and express them separately in different organelles (such as the chloroplast and peroxisome) or in the cytosol. Therefore, genetic engineers may have to target multiple duplicated genes to manipulate expression in different cell parts. To get around this problem, <a href="http://www.jove.com/video/51234/">Mattozzi et al.</a> use a novel method for multiple-organelle targeting that takes advantage of alternatively spliced mRNAs, which are sent to different locations in the cell. The targeting constructs are designed using Gibson assembly, a one-step cloning method, and then delivered using a gene gun. This technique offers great flexibility in terms of genetic design, and can be applied to large-scale selective genetic engineering in agriculture or the environment.
In the <a href="http://www.jove.com/chem">Chemistry</a> section we feature an article that uses modern chemical techniques to examine two bright blue pigments that were synthesized in ancient societies. Calcium copper tetrasilicate, commonly known as Egyptian blue, is thought to be the first synthetic pigment and used in Egypt as early the 4th Dynasty (ca. 2500 BC), the height of the pyramid-building era. Barium copper tetrasilicate, also known as Han blue, was later developed in China during the beginning of the Zhou Dynasty, or the height of the bronze age (1045-771 BC). Nowadays, scientists are interested in the strong near infrared (NIR) emission properties of these two pigments, which can be utilized for optical sensors. Related to this application is the ability to exfoliate copper tetrasilicates into 2D monolayers, or nanosheets. <a href="http://www.jove.com/video/51686">Johnson-McDaniel and Salguero</a> demonstrate how to synthesize these two copper tetrasilicate pigments and exfoliate them into monolayer nanosheets. Egyptian blue spontaneously exfoliates into monolayers when stirred in hot water, whereas Han blue requires ultrasonication in organic solvents. Our authors show that both the bulk and nanosheet forms of the two pigments are strong near infrared emitters with potential applications ranging from security inks to biomedical imaging.
You've just had a sneak peek of the <a href="http://www.jove.com/archive/86">April 2014 issue</a> of JoVE. Visit the website to see the full-length articles, plus many more, in <a href="http://www.jove.com/">JoVE: The Journal of Visualized Experiments</a>.

Watch this Scientific Journal Video about April 2014: This Month in JoVE - Bioengineering for ACL Tears, Neuroscience behind Insect Locomotion, Improved Genetic Engineering of Crops, and Synthesizing 

April 2014: This Month in JoVE - Bioengineering for ACL Tears, Neuroscience behind Insect Locomotion, Improved Genetic Engineering of Crops, and Synthesizing Secret Ink

Here's a look at what's coming up in the <a href="http://www.jove.com/archive/86">April 2014</a> issue of JoVE.

Here's a look at what's coming up in the April 2014 issue of JoVE: The Journal of Visualized Experiments.
In the Bioengineering section we have a video ...

april-2014-this-month-jove-bioengineering-for-acl-tears-neuroscience

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2.0K ビュー.  Massachusetts Eye and Ear. Here's a look at what's coming up in the April 2014 issue of JoVE.

ビデオ: April 2014: This Month in JoVE - Bioengineering for ACL Tears, Neuroscience behind Insect Locomotion, Improved Genetic Engineering of Crops, and Synthesizing Secret Ink

genetic engineering of an unconventional yeast for renewable biofuel and biochemical production

Genetic Engineering of an Unconventional Yeast for Renewable Biofuel and Biochemical Production

We herein report methods on the molecular genetic manipulation of the Yarrowia lipolytica Po1g strain for improved gene deletion efficiency. The resulting engineered Y. lipolytica strains have potential applications in biofuel and biochemical...

universal and efficient electroporation protocol for genetic engineering of gastrointestinal organoids

Universal and Efficient Electroporation Protocol for Genetic Engineering of Gastrointestinal Organoids

This protocol describes an efficient electroporation method for the transfection of four different gastrointestinal organoid entities with larger plasmids (to the extent of 10 kB). It can be performed within one day and does not need extensive preparation or special, cost-intensive electroporation...

electroporation of agrobacterium tumefaciens for genetic engineering of microalgal cells

Electroporation of  Agrobacterium tumefaciens  for Genetic Engineering of Microalgal Cells  

Electroporation of Agrobacterium tumefaciens for Genetic Engineering of Microalgal Cells

establishing an octopus ecosystem for biomedical and bioengineering research

Establishing an Octopus Ecosystem for Biomedical and Bioengineering Research

Understanding the unique physiological and anatomical structures of octopuses can greatly impact biomedical research. This guide demonstrates how to set-up and maintain a marine environment to accommodate this species and includes state-of-the-art imaging and analytical approaches to visualize octopus' nervous system anatomy and...

isolation, culture, and genetic engineering of mammalian primary pigment epithelial cells for non-viral gene therapy

Isolation, Culture, and Genetic Engineering of Mammalian Primary Pigment Epithelial Cells for Non-Viral Gene Therapy

Here, a protocol to isolate and transfect primary iris and retinal pigment epithelial cells from various mammals (mice, rat, rabbit, pig, and bovine) is presented. The method is ideally suited to study ocular gene therapy approaches in various set-ups for ex vivo&#160;analyses and&#160;in vivo studies transferable to...

antimicrobial characterization of advanced materials for bioengineering applications

Antimicrobial Characterization of Advanced Materials for Bioengineering Applications

We present a protocol for the antimicrobial characterization of advanced materials. Here, the antimicrobial activity on material surfaces is measured by two methods that complement each other: one is based on the agar disk diffusion test, and the other is a standard procedure based on the ISO 22196:2007...

preparation of electrocompetent agrobacterium tumefaciens culture for microalgal genetic engineering

Preparation of Electrocompetent  Agrobacterium tumefaciens  Culture for Microalgal Genetic Engineering  

Preparation of Electrocompetent Agrobacterium tumefaciens Culture for Microalgal Genetic Engineering

genetic engineering of dictyostelium discoideum cells based on selection and growth on bacteria

Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria

Dictyostelium discoideum is a popular model organism to study complex cellular processes such as cell migration, endocytosis, and development. The utility of the organism is dependent on the feasibility of genetic manipulation. Here, we present methods to transfect Dictyostelium discoideum cells that overcome existing limitations of culturing cells in liquid...

Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria

on-chip octanol-assisted liposome assembly for bioengineering

On-Chip Octanol-Assisted Liposome Assembly for Bioengineering

The present protocol describes octanol-assisted liposome assembly (OLA), a microfluidic technique to generate biocompatible liposomes. OLA produces monodispersed, micron-sized liposomes with efficient encapsulation, allowing immediate on-chip experimentation. This protocol is anticipated to be particularly suitable for synthetic biology and synthetic cell...

agrobacterium tumefaciens-mediated genetic engineering of green microalgae, chlorella vulgaris

Author Spotlight: Optimized Transformation Protocol for Chlorella vulgaris Using Agrobacterium tumefaciens 

This protocol outlines the utilization of Agrobacterium tumefaciens-mediated transformation (AMT) for integrating gene(s) of interest into the nuclear genome of the green microalgae Chlorella vulgaris, leading to the production of stable...

Agrobacterium tumefaciens-Mediated Genetic Engineering of Green Microalgae, Chlorella vulgaris

bioengineering of humanized bone marrow microenvironments in mouse and their visualization by live imaging

Bioengineering of Humanized Bone Marrow Microenvironments in Mouse and Their Visualization by Live Imaging

A method to create and live-image different humanized bone-marrow niches in mice is presented. Based on the supportive niche created by human mesenchymal cells, the addition of human endothelial cells induces the formation of human vessels, while the addition of rhBMP-2 induces the formation of human-mouse chimeric mature bone...

genetic engineering of primary mouse intestinal organoids using magnetic nanoparticle transduction viral vectors for frozen sectioning

Genetic Engineering of Primary Mouse Intestinal Organoids Using Magnetic Nanoparticle Transduction Viral Vectors for Frozen Sectioning

We describe step-by-step instructions to: 1) efficiently engineer intestinal organoids using magnetic nanoparticles for lenti- or retroviral transduction, and, 2) generate frozen sections from engineered organoids. This approach provides a powerful tool to efficiently alter gene expression in organoids for investigation of downstream...

an improved assay and tools for measuring mechanical nociception in drosophila larvae

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

The goal of this protocol is to show how to perform an improved assay for mechanical nociception in Drosophila larvae. We use the assay here to demonstrate that mechanical hypersensitivity (allodynia and hyperalgesia) exists in Drosophila...

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

bioengineering human microvascular networks in immunodeficient mice

Bioengineering Human Microvascular Networks in Immunodeficient Mice

Here, we describe a methodology to deliver human cord blood-derived endothelial colony-forming cells (ECFCs) and bone marrow-derived mesenchymal stem cells (MSCs), embedded in a collagen/fibronectin gel, subcutaneously into immunodeficient mice. This cell/gel combination generates a human vascular network that connects with the mouse...

battery of behavioral tests assessing general locomotion, muscular strength, and coordination in mice

Battery of Behavioral Tests Assessing General Locomotion, Muscular Strength, and Coordination in Mice

Behavioral testing is used in pre-clinical trials to assess the phenotypic effects of a disease or treatment on an animal's wellbeing. In order to globally assess motor functioning, we selected tests for general locomotion, muscular strength, and coordination: the open field test, the mesh test, and the rotarod test, respectively.

adaptable angled stereotactic approach for versatile neuroscience techniques

Adaptable Angled Stereotactic Approach for Versatile Neuroscience Techniques

Described here is a stereotactic procedure that can target challenging and difficult-to-reach brain regions (due to spatial limitations) using an angled coronal approach. This protocol is adaptable to both mouse and rat models and can be applied to diverse neuroscientific applications, including cannula implantation and microinjections of viral...

an improved method for accurate and rapid measurement of flight performance in drosophila

An Improved Method for Accurate and Rapid Measurement of Flight Performance in Drosophila

Here we describe a method for rapid and accurate measurement of flight performance in Drosophila, enabling high-throughput...

An Improved Method for Accurate and Rapid Measurement of Flight Performance in Drosophila

Stable Gene Integration in &lt;em&gt;Chlorella vulgaris&lt;/em&gt; Using &lt;em&gt;Agrobacterium tumefaciens&lt;/em&gt;

https://cloudfront.jove.com/CDNSource/teasers/65382_b.jpg

high-resolution video tracking of locomotion in adult drosophila melanogaster

High-Resolution Video Tracking of Locomotion in Adult Drosophila Melanogaster

The study of complex locomotor behavior in Drosophila melanogaster is dependent upon the ability to quantify changes in a given fly's movement.  This article demonstrates how to do this using a high-resolution tracking...

insect-controlled robot: a mobile robot platform to evaluate the odor-tracking capability of an insect

Insect-controlled Robot: A Mobile Robot Platform to Evaluate the Odor-tracking Capability of an Insect

The capability to localize an odor source is necessary for insect survival and is expected to be applicable to artificial odor-tracking. The insect-controlled robot is driven by an actual silkmoth and enables us to evaluate the odor-tracking capability of insects through a robotic...

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 ...