<h2 class="jove_title"><a href="https://www.jove.com/video/52317">Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 1. Biosafety Level 4 Suit Laboratory Suite Entry and Exit Procedures</a></h2>Krisztina Janosko1, Michael R. Holbrook1, Ricky Adams1, Jason Barr1, Laura Bollinger1, Je T'aime Newton2, Corrie Ntiforo2, Linda Coe1, Jiro Wada1, Daniela Pusl1, Peter B. Jahrling1, Jens H. Kuhn1, Matthew G. Lackemeyer11Integrated Research Facility at Frederick, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), 2Environmental Health and Safety, Biological and Chemical Safety Program, University of Texas Medical BranchAlthough researchers are generally knowledgeable about procedures and safety precautions required for biosafety level 1 or 2 (BSL-1/2) experiments, they may not be familiar with experimental procedures in BSL-4 suit laboratories. This article provides a detailed visual demonstration of BSL-4 suit laboratory systems check, laboratory entry, movement, and exit procedures.<h2 class="jove_title"><a href="https://www.jove.com/video/53600">Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 2. General Practices</a></h2>Steven Mazur, Michael R. Holbrook, Tracey Burdette, Nicole Joselyn, Jason Barr, Daniela Pusl, Laura Bollinger, Linda Coe, Peter B. Jahrling, Matthew G. Lackemeyer, Jiro Wada, Jens H. Kuhn, Krisztina JanoskoIntegrated Research Facility at Frederick, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH)Performing viral assays in a BSL-4 laboratory is more involved compared to work in a BSL-2 laboratory due to required additional safety precautions. Here, we present an overview of practices and procedures used inside a BSL-4 laboratory illustrating proper Class II biosafety cabinet usage, waste management/disposal, and sample removal.<h2 class="jove_title"><a href="https://www.jove.com/video/53601">Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 4. Medical Imaging Procedures</a></h2>Russell Byrum, Lauren Keith, Christopher Bartos, Marisa St. Claire, Matthew G. Lackemeyer, Michael R. Holbrook, Krisztina Janosko, Jason Barr, Daniela Pusl, Laura Bollinger, Jiro Wada, Linda Coe, Lisa E. Hensley, Peter B. Jahrling, Jens H. Kuhn, Margaret R. LentzIntegrated Research Facility at Frederick, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH)Here, we present an overview of the preparation and animal handling procedures required to safely perform medical imaging in an animal biosafety level 4 laboratory. Computed tomography of a mock-infected guinea pig illustrates these procedures that may be used to evaluate the disease caused by a high consequence pathogen.<h2 class="jove_title"><a href="https://www.jove.com/video/53602">Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 3. Aerobiology</a></h2>J. Kyle Bohannon, Krisztina Janosko, Michael R. Holbrook, Jason Barr, Daniela Pusl, Laura Bollinger, Linda Coe, Lisa E. Hensley, Peter B. Jahrling, Jiro Wada, Jens H. Kuhn, Matthew G. LackemeyerIntegrated Research Facility at Frederick, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH)As high-consequence pathogens can potentially infect subjects through airborne particles, aerobiology has been increasingly applied in pathogenesis research and medical countermeasure development. We present a detailed visual demonstration of aerobiology procedures during an aerosol challenge in nonhuman primates in an animal biosafety level 4 maximum containment environment.<h2 class="jove_title"><a href="https://www.jove.com/video/54248">Investigating the 'Uncatchable Smile' in Leonardo da Vinci's La Bella Principessa: A Comparison with the Mona Lisa and Pollaiuolo's Portrait of a Girl</a></h2>Alessandro Soranzo, Michelle NewberryDepartment of Psychology, Sociology and Politics, Sheffield Hallam UniversityThis paper discusses the methodology used to reveal the &#39;Uncatchable Smile&#39; illusion in Leonardo da Vinci&#39;s La Bella Principessa portrait. A combination of three methods was used (inter-observation, structured interviews, and psychophysical experiments), which led to an investigation that shaped itself without prior beliefs, thus reducing potential researcher bias.<h2 class="jove_title"><a href="https://www.jove.com/video/54553">The Use of Induced Somatic Sector Analysis (ISSA) for Studying Genes and Promoters Involved in Wood Formation and Secondary Stem Development</a></h2>Antanas Spokevicius1, Lynette Taylor1, Emma Melder1, Kim Van Beveren1, Josquin Tibbits2, Nicky Creux3,4, Gerd Bossinger11School of Ecosystem and Forest Sciences, Faculty of Science, The University of Melbourne, 2Victorian AgriBiosciences Centre, La Trobe University R&amp;D Park, 3College of Biological Sciences, Department of Plant Biology, University of California, Davis, 4Department of Genetics, Forestry and Agricultural Biotechnology Institute (FABI), University of PretoriaHere we present a protocol that facilitates the medium to high throughput functional characterization of gene and promoter constructs in tree secondary stem tissue within comparatively short time frames. It is efficient, easy to use and widely applicable to a range of tree species.

october 2016: this month in jove

Here's a look at what's coming up in the October 2016 issue of <a href="http://www.jove.com/">JoVE: The Journal of Visualized Experiments</a>.
In <a href="http://www.jove.com/i2">Immunology and Infection</a>, we showcase a series of four videos detailing safety and logistical procedures for working in an Animal Biosafety Level 4, or (A)BSL-4, laboratory. The <a href="http://www.jove.com/video/52317/safety-precautions-operating-procedures-an-bsl-4-laboratory-1?status=a54323k">first</a> of this group, filmed at the NIH Integrated Research Facility at Fort Detrick, demonstrates the safe entry and exit procedures for work inside an (A)BSL-4 suit laboratory suite. Inhalation studies of high-consequence pathogens can simulate natural aerosol transmission, or allow researchers to investigate outcomes of intentional pathogenic aerosol releases. In the <a href="http://www.jove.com/video/53602/safety-precautions-operating-procedures-an-bsl-4-laboratory-3">second</a> of this series, the authors walk through the safe operation of aerobiology chambers for maximum containment level pathogens, such as the Ebola virus.
The challenge of carrying out <a href="http://www.jove.com/video/53601/safety-precautions-operating-procedures-an-bsl-4-laboratory-4-medical">medical imaging</a> in a high biosafety environment is the focus of the third of these releases. Here, our Authors detail how to prepare animals infected with high-consequence pathogens for noninvasive medical imaging, whilst ensuring that the equipment remains easily accessed and free from contamination. In the <a href="http://www.jove.com/video/53600/safety-precautions-operating-procedures-an-bsl-4-laboratory-2-general">final video</a> of this miniseries, we take an in-depth look at the extra precautions and procedures involved in performing viral assays in a Class II biosafety cabinet in a BSL-4 environment. As a whole, these four releases are a valuable library for researchers handling challenging and potentially harmful biological materials.
Woody plants and secondary tree stems are key habitats, as well as being of great cultural and commercial importance. Understanding stem growth and wood formation is therefore an important topic for tree production, conservation, and preservation. In <a href="http://www.jove.com/">JoVE Genetics</a> this month, <a href="http://www.jove.com/video/54553/the-use-induced-somatic-sector-analysis-issa-for-studying-genes">Spokevicius et al</a> (our Authors) describe a method to create transgenic somatic tissue sectors directly in the living secondary stem of woody plants. This versatile method can facilitate rapid functional characterization of genes of interest, be utilized in a range of tree species, and test multiple genes and promoters at a high throughput level.
Arguably one of the most recognizable paintings of all time, the Mona Lisa is famous for the subject's elusive smile and indefinable expression. In <a href="http://www.jove.com/behavior">JoVE Behavior</a>, <a href="http://www.jove.com/video/54248/investigating-uncatchable-smile-leonardo-da-vinci-s-la-bella">Soranzo and Newbury</a> (our Authors) demonstrate how a similar visual illusion-dubbed the "uncatchable smile" because of it's tendency to disappear as the observer tries to catch it-was discovered in La Bella Principessa, also by Da Vinci. Using a combination of methods including interobservation, psychophysical experiments, and structured interviews, the authors reveal that the ambiguity in La Bella Principessa's expression is attributed to a visual illusion at the mouth of the subject similar to that observed in the Mona Lisa.
You've just had a sneak peek of the October 2016 issue of <a href="http://www.jove.com/">JoVE</a>. 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 October 2016: This Month in JoVE at JoVE.com

October 2016: This Month in JoVE

Here's a look at what's coming up in the October 2016 issue of JoVE: The Journal of Visualized Experiments.
In Immunology and Infection, we showcase a ...

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1.6K ビュー.  JoVE Content Production. October 2016: This Month in JoVE

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intraoperative assessment of resection margins in oral cavity cancer: this is the way

Intraoperative Assessment of Resection Margins in Oral Cavity Cancer: This is the Way

The goal of this protocol is to provide a clear overview of specimen-driven intraoperative assessment of resection margins. It is encouraged to implement this protocol to improve patient care at other...

measuring composition of cd95 death-inducing signaling complex and processing of procaspase-8 in this complex

Measuring Composition of CD95 Death-Inducing Signaling Complex and Processing of Procaspase-8 in this Complex

Here, an experimental workflow is presented that enables the detection of caspase-8 processing directly at the death-inducing signaling complex (DISC) and determines the composition of this complex. This methodology has broad applications, from unraveling the molecular mechanisms of cell death pathways to the dynamic modeling of apoptosis...

experience is instrumental in tuning a link between language and cognition: evidence from 6-  to 7- month-old infants' object categorization

Experience is Instrumental in Tuning a Link Between Language and Cognition: Evidence from 6-  to 7- Month-Old Infants' Object Categorization

At 3-4 months, listening to human and nonhuman primate vocalizations boosts infants&#39; cognition; by 6 months, only human vocalizations exert this cognitive advantage. We describe an exposure manipulation that reveals the powerful shaping role of experience as infants specify which sounds to link to cognition and which to tune...

vitrification of in vitro matured oocytes collected from adult and prepubertal ovaries in sheep

Vitrification of In Vitro Matured Oocytes Collected from Adult and Prepubertal Ovaries in Sheep

The protocol aims at providing a standard method for the vitrification of adult and juvenile sheep oocytes. It includes all the steps from the preparation of the in vitro maturation media to the post-warming culture. Oocytes are vitrified at the MII stage using Cryotop to ensure the minimum essential volume.

in vivo chronic two-photon imaging of microglia in the mouse hippocampus

In Vivo Chronic Two-Photon Imaging of Microglia in the Mouse Hippocampus

This paper describes a method for chronic in vivo observation of the resting microglia in the mouse hippocampal CA1 using precisely controlled surgery and two-photon...

In Vivo Chronic Two-Photon Imaging of Microglia in the Mouse Hippocampus

transcranial direct current stimulation (tdcs) in mice

Transcranial Direct Current Stimulation tDCS in Mice

Transcranial direct current stimulation (tDCS) is a therapeutic technique proposed to treat psychiatric diseases. An animal model is essential for understanding the specific biological alterations evoked by tDCS. This protocol describes a tDCS mouse model that uses a chronically implanted...

Transcranial Direct Current Stimulation (tDCS) in Mice

in vivo targeting of neural progenitor cells in ferret neocortex by in utero electroporation

In Vivo Targeting of Neural Progenitor Cells in Ferret Neocortex by In Utero Electroporation

Presented here is a protocol to perform genetic manipulation in the embryonic ferret brain using in utero electroporation. This method allows for targeting of neural progenitor cells in the neocortex in...

adjunctive diode laser therapy and probiotic lactobacillus therapy in the treatment of periodontitis and peri-implant disease

Adjunctive Diode Laser Therapy and Probiotic Lactobacillus Therapy in the Treatment of Periodontitis and Peri-Implant Disease

This article describes two protocols: 1) adjunctive diode laser therapy for treating periodontitis and 2) probiotic Lactobacillus therapy for treating peri-implant disease, with emphasis on the laser usage mode (inside or outside pocket), laser application regimen (single or multiple sessions), and a probiotic protocol of professional and home...

Adjunctive Diode Laser Therapy and Probiotic Lactobacillus Therapy in the Treatment of Periodontitis and Peri-Implant Disease

implantation of a cranial window for repeated in vivo imaging in awake mice

Implantation of a Cranial Window for Repeated In Vivo Imaging in Awake Mice

Presented here is a protocol for the implantation of a chronic cranial window for the longitudinal imaging of brain cells in awake, head-restrained...

Implantation of a Cranial Window for Repeated In Vivo Imaging in Awake Mice

noninvasive intratracheal lipopolysaccharide instillation in mice

Noninvasive Intratracheal Lipopolysaccharide Instillation in Mice

Here, we propose a protocol for intratracheal lipopolysaccharide (LPS) delivery via noninvasive oropharyngeal endotracheal intubation. This method minimizes the trauma of the surgical procedure for the animal and accurately delivers LPS to the trachea and then to the lungs.

aav deployment of enhancer-based expression constructs in vivo in mouse brain

AAV Deployment of Enhancer-Based Expression Constructs In Vivo in Mouse Brain

This protocol describes a novel rAAV-based transient enhancer-reporter assay. This assay can be used to induce enhancer-driven expression in vivo in the mouse...

AAV Deployment of Enhancer-Based Expression Constructs In Vivo in Mouse Brain

assaying for inorganic polyphosphate in bacteria

Assaying for Inorganic Polyphosphate in Bacteria

We describe a simple method for rapid quantification of inorganic polyphosphate in diverse bacteria, including Gram-negative, Gram-positive, and mycobacterial...

post-myocardial infarction heart failure in closed-chest coronary occlusion/reperfusion model in g&#246;ttingen minipigs and landrace pigs

Post-Myocardial Infarction Heart Failure in Closed-chest Coronary Occlusion/Reperfusion Model in G&amp;#246;ttingen Minipigs and Landrace Pigs

The overall goal of the current study is to present the techniques of induction of myocardial infarction (MI) and post-myocardial infarction heart failure (post-MI HF) in closed-chest, adult G&#246;ttingen minipigs and the characterization of post-MI HF model in G&#246;ttingen minipigs as compared to Landrace pigs.

Post-Myocardial Infarction Heart Failure in Closed-chest Coronary Occlusion/Reperfusion Model in G&#246;ttingen Minipigs and Landrace Pigs

preparation of gushukang (gsk) granules for in vivo and in vitro experiments

Preparation Of Gushukang GSK Granules for In Vivo and In Vitro Experiments

This article provides a detailed protocol for preparing a working solution of Gushukang granules for animal studies and GSK granule containing serum for in vitro experiments. This protocol can be applied to pharmacological investigations of herbal medicines as well as prescriptions for both in vivo and in vitro...

Preparation Of Gushukang (GSK) Granules for In Vivo and In Vitro Experiments

in vivo assessment of microtubule dynamics and orientation in caenorhabditis elegans neurons

In vivo Assessment of Microtubule Dynamics and Orientation in Caenorhabditis elegans Neurons

A protocol for imaging the dynamic microtubules in vivo using fluorescently labeled End binding protein has been presented. We described the methods to label, image, and analyze the dynamic microtubules in the Posterior lateral microtubule (PLM) neuron of C.

In vivo Assessment of Microtubule Dynamics and Orientation in Caenorhabditis elegans Neurons

genotyping and quantification of in situ hybridization staining in zebrafish

Genotyping and Quantification of In Situ Hybridization Staining in Zebrafish

Gene editing technologies have enabled researchers to generate zebrafish mutants to investigate gene function with relative ease. Here, we provide a guide to perform parallel embryo genotyping and quantification of in situ hybridization signals in zebrafish. This unbiased approach provides greater accuracy in phenotypical analyses based on in situ...

assessment of the efficacy of an osteopathic treatment in infants with biomechanical impairments to suckling

Assessment of the Efficacy of An Osteopathic Treatment in Infants with Biomechanical Impairments to Suckling

Osteopathy is an emerging field of clinical research. Here we present a protocol to assess the efficacy of an osteopathic intervention coupled with lactation consultation, in infants with biomechanical issues impeding...

determining gender-based differences in retinal and choroidal thickness in underweight individuals via swept-source optical coherence tomography

Author Spotlight: Determining Gender-Based Differences in Retinal and Choroidal Thickness in Underweight Individuals via Swept-Source Optical Coherence Tomography

Here, swept-source optical coherence tomography (SS-OCT) is used to compare retinal and choroidal thickness in adults with and without malnutrition, contributing to a better understanding of the pathogenesis of ocular diseases in malnourished...

Determining Gender-Based Differences in Retinal and Choroidal Thickness in Underweight Individuals via Swept-Source Optical Coherence Tomography

in situ immunofluorescent staining of autophagy in muscle stem cells

In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells

Active autophagy is associated with productive muscle regeneration, which is essential for Muscle Stem Cell (MuSC) activation. Here, we provide a protocol for the in situ detection of LC3, an autophagy marker in MyoD-positive MuSCs of muscle tissue sections from control and injured...

In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells

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 ...