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The Journal of Visualized Experiments (JoVE) is a peer reviewed, PubMed-indexed video journal. Our mission is to increase the productivity of scientific research.

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This month in JoVE May, 2013

The Journal of Visualized Experiments (JoVE) is a peer reviewed, PubMed-indexed video journal. Our mission is to increase the productivity of scientific research.

May 2013: This Month in JoVE

Wendy Chao¹, Aaron Kolski-Andreaco²

¹Department of Ophthalmology, Massachusetts Eye and Ear, ²JoVE Content Production

A Caenorhabditis elegans Model System for Amylopathy Study

Zhibing Duan, Federico Sesti

Department of Neuroscience and Cell Biology, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey

A Practical and Novel Method to Extract Genomic DNA from Blood Collection Kits for Plasma Protein Preservation

Jon Waters¹, Vishal Dhere¹, Adam Benjamin¹, Arvind Sekar¹, Archana Kumar¹, Sampath Prahalad², David T. Okou¹, Subra Kugathasan¹

¹Division of Gastroenterology, Hepatology and Nutrition, Department of Pediatrics, Emory University School of Medicine and Children's Health Care of Atlanta, ²Division of Rheumatology, Department of Pediatrics, Emory University School of Medicine and Children's Health Care of Atlanta

Live Cell Cycle Analysis of Drosophila Tissues using the Attune Acoustic Focusing Cytometer and Vybrant DyeCycle Violet DNA Stain

Kerry Flegel*, Dan Sun*, Olga Grushko*, Yiqin Ma, Laura Buttitta

Molecular, Cellular and Developmental Biology, University of Michigan

* These authors contributed equally

Measuring Spatially- and Directionally-varying Light Scattering from Biological Material

Todd Alan Harvey¹, Kimberly S. Bostwick^2,3, Steve Marschner⁴

¹Department of Biomedical Science, Cornell University, ²Department of Ecology and Evolutionary Biology, Cornell University, ³Cornell University Museum of Vertebrates, ⁴Department of Computer Science, Cornell University

Untargeted Metabolomics from Biological Sources Using Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)

Nathaniel W. Snyder, Maya Khezam, Clementina A. Mesaros, Andrew Worth, Ian A. Blair

Centers for Cancer Pharmacology and Excellence in Environmental Toxicology, Department of Pharmacology, University of Pennsylvania

Matrix-assisted Autologous Chondrocyte Transplantation for Remodeling and Repair of Chondral Defects in a Rabbit Model

Markus T. Berninger^1,2, Gabriele Wexel³, Ernst J. Rummeny², Andreas B. Imhoff¹, Martina Anton³, Tobias D. Henning*^2,4, Stephan Vogt*¹

¹Department of Orthopaedic Sports Medicine, Klinikum rechts der Isar der Technischen Universität München, ²Department of Radiology, Klinikum rechts der Isar der Technischen Universität München, ³Institute of Experimental Oncology and Therapy Research, Klinikum rechts der Isar der Technischen Universität München, ⁴Department of Radiology, Uniklinik Köln

* These authors contributed equally

Treatment of Osteochondral Defects in the Rabbit's Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

Markus T. Berninger^1,2, Gabriele Wexel³, Ernst J. Rummeny², Andreas B. Imhoff¹, Martina Anton³, Tobias D. Henning*^2,4, Stephan Vogt*¹

¹Department of Orthopaedic Sports Medicine, Klinikum rechts der Isar der Technischen Universität München, ²Department of Radiology, Klinikum rechts der Isar der Technischen Universität München, ³Institute of Experimental Oncology and Therapy Research, Klinikum rechts der Isar der Technischen Universität München, ⁴Department of Radiology, Uniklinik Köln

* These authors contributed equally

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Most Recent Video Articles

Treatment of Osteochondral Defects in the Rabbit's Knee<span style='display: none;'> Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots</span>…

Treatment of Osteochondral Defects in the Rabbit's Knee…
Published Yesterday

An experimental technique for the treatment of osteochondral defects in the rabbit's knee joint is described. The implantation of allogeneic mesenchymal stem cells into osteochondral defects provides a promising development in the field of tissue engineering. The preparation of fibrin-cell-clots in vitro offers a standardized method for implantation.

Matrix-assisted Autologous Chondrocyte Transplantation for<span style='display: none;'> Remodeling and Repair of Chondral Defects in a Rabbit Model</span>…

Matrix-assisted Autologous Chondrocyte Transplantation for…
Published Yesterday

An experimental technique for the treatment of chondral defects in the rabbit's knee joint is described. The implantation of autologous chondrocytes seeded on a matrix is a well-accepted method for the remodeling and repair of articular cartilage lesions providing satisfying long-term results. Matrix-assisted autologous chondrocyte transplantation (MACT) offers a standardized and clinically established implantation method.

Untargeted Metabolomics from Biological Sources Using<span style='display: none;'> Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)</span>…

Untargeted Metabolomics from Biological Sources Using…
Published 5/20/2013

Untargeted metabolomics provides a hypothesis generating snapshot of a metabolic profile. This protocol will demonstrate the extraction and analysis of metabolites from cells, serum, or tissue. A range of metabolites are surveyed using liquid-liquid phase extraction, microflow ultraperformance liquid chromatography/high-resolution mass spectrometry (UPLC-HRMS) coupled to differential analysis software.

Measuring Spatially- and Directionally-varying Light<span style='display: none;'> Scattering from Biological Material</span>…

Measuring Spatially- and Directionally-varying Light Scattering from Biological Material…
Published 5/20/2013

We present a non-destructive method for sampling spatial variation in the direction of light scattered from structurally complex materials. By keeping the material intact, we preserve gross-scale scattering behavior, while concurrently capturing fine-scale directional contributions with high-resolution imaging. Results are visualized in software at biologically-relevant positions and scales.

Live Cell Cycle Analysis of <em>Drosophila</em> Tissues<span style='display: none;'> using the Attune Acoustic Focusing Cytometer and Vybrant DyeCycle Violet DNA Stain</span>…

Live Cell Cycle Analysis of Drosophila Tissues…
Published 5/19/2013

A protocol for cell cycle analysis of live Drosophila tissues using the Attune Acoustic Focusing Cytometer is described. This protocol simultaneously provides information about relative cell size, cell number, DNA content and cell type via lineage tracing or tissue specific expression of fluorescent proteins in vivo.

A Practical and Novel Method to Extract Genomic DNA from<span style='display: none;'> Blood Collection Kits for Plasma Protein Preservation</span>…

A Practical and Novel Method to Extract Genomic DNA from…
Published 5/18/2013

We are describing a new method of isolating genomic DNA from whole blood collected for plasma/serology. After plasma collection, the compacted blood is usually discarded. Our novel method represents a significant improvement over existing methods and makes DNA and plasma available from a single collection, without requesting additional blood.

Evaluation of Respiratory System Mechanics in Mice using<span style='display: none;'> the Forced Oscillation Technique</span>…

Evaluation of Respiratory System Mechanics in Mice using the Forced Oscillation Technique…
Published 5/15/2013

The present protocol provides a detailed step-by-step description of the procedures required to execute measurements of respiratory system mechanics as well as the assessment of airway responsiveness to inhaled methacholine in mice using the forced oscillation technique (flexiVent; SCIREQ Inc, Montreal, Qc, Canada).

Protein Transfection of Mouse Lung

Protein Transfection of Mouse Lung
Published 5/15/2013

Transgenic mice or viral vectors have been used to increase protein expression within the lung. However, these techniques are time-consuming, technically challenging and have off-target effects that can confound results. Our protein transfection protocol uses a lipid based transfection reagent and an ultrafine microsprayer to uniformly deliver active protein to lung cells.

A <em>Caenorhabditis elegans</em> Model System for<span style='display: none;'> Amylopathy Study</span>…

A Caenorhabditis elegans Model System for Amylopathy Study…
Published 5/17/2013

We describe methods to study aspects of amylopathies in the worm C. elegans. We show how to construct worms expressing human Aβ₄₂ in neurons and how to test their function in behavioral assays. We further show how to obtain primary neuronal cultures that can be used for pharmacological testing.

Implementing Dynamic Clamp with Synaptic and Artificial<span style='display: none;'> Conductances in Mouse Retinal Ganglion Cells</span>…

Implementing Dynamic Clamp with Synaptic and Artificial…
Published 5/16/2013

This video article illustrates the set-up, the procedures to patch cell bodies and how to implement dynamic clamp recordings from ganglion cells in whole-mount mouse retinae. This technique allows the investigation of the precise contribution of excitatory and inhibitory synaptic inputs, and their relative magnitude and timing to neuronal spiking.

Direct Intraventricular Delivery of Drugs to the Rodent<span style='display: none;'> Central Nervous System</span>…

Direct Intraventricular Delivery of Drugs to the Rodent Central Nervous System…
Published 5/12/2013

We describe a method to target drugs to the central nervous system by either implanting a catheter or performing a bolus injection into the right lateral ventricle in mice. We focus specifically on the delivery of antisense oligonucleotides. This technique is readily adaptable to other drugs and to rats.

F<sub>1</sub>F<sub>O</sub> ATPase Vesicle Preparation and<span style='display: none;'> Technique for Performing Patch Clamp Recordings of Submitochondrial Vesicle Membranes</span>…

F₁F_O ATPase Vesicle Preparation and…
Published 5/04/2013

A method to isolate submitochondrial vesicles enriched in F1FO ATP synthase complexes from rat brain is described. These vesicles allow the study of the activity of F1FO ATPase complex and its modulation using the technique of patch clamp recording.

A Simple Protocol for Platelet-mediated Clumping of<span style='display: none;'> Plasmodium falciparum-infected Erythrocytes in a Resource Poor Setting</span>…

A Simple Protocol for Platelet-mediated Clumping of…
Published 5/16/2013

This method investigates the platelet-mediated clumping phenotype of Plasmodium falciparum-infected erythrocytes (pRBC) in clinical isolates. This is performed by isolating and co-incubating platelet-rich plasma and a suspension of pRBC.

Accelerated Type 1 Diabetes Induction in Mice by Adoptive<span style='display: none;'> Transfer of Diabetogenic CD4+ T Cells</span>…

Accelerated Type 1 Diabetes Induction in Mice by Adoptive Transfer of Diabetogenic CD4+ T Cells…
Published 5/06/2013

We provide a reproducible method to induce type 1 diabetes (T1D) in mice within two weeks by the adoptive transfer of islet antigen-specific, primary CD4+ T cells.

Oral Transmission of <em>Listeria monocytogenes</em> in<span style='display: none;'> Mice via Ingestion of Contaminated Food</span>…

Oral Transmission of Listeria monocytogenes in…
Published 5/06/2013

This paper describes a novel method for oral infection of mice using Listeria monocytogenes-contaminated food. The protocol can readily be adapted for use with other food borne bacterial pathogens.

Trans-vivo Delayed Type Hypersensitivity Assay for Antigen<span style='display: none;'> Specific Regulation</span>…

Trans-vivo Delayed Type Hypersensitivity Assay for Antigen Specific Regulation…
Published 5/02/2013

We describe a valuable diagnostic assay that could potentially be used to decide the withdrawal of immunosuppression after transplant without elevated risk of graft rejection. The assay uses the principles of Delayed Type Hypersensitivity and provides accurate assessment of both donor specific effector and regulatory immune responses mounted by recipients.

Treatment of Osteochondral Defects in the Rabbit's Knee<span style='display: none;'> Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots</span>…

Treatment of Osteochondral Defects in the Rabbit's Knee…
Published Yesterday

An experimental technique for the treatment of osteochondral defects in the rabbit's knee joint is described. The implantation of allogeneic mesenchymal stem cells into osteochondral defects provides a promising development in the field of tissue engineering. The preparation of fibrin-cell-clots in vitro offers a standardized method for implantation.

Matrix-assisted Autologous Chondrocyte Transplantation for<span style='display: none;'> Remodeling and Repair of Chondral Defects in a Rabbit Model</span>…

Matrix-assisted Autologous Chondrocyte Transplantation for…
Published Yesterday

An experimental technique for the treatment of chondral defects in the rabbit's knee joint is described. The implantation of autologous chondrocytes seeded on a matrix is a well-accepted method for the remodeling and repair of articular cartilage lesions providing satisfying long-term results. Matrix-assisted autologous chondrocyte transplantation (MACT) offers a standardized and clinically established implantation method.

<em>In vitro</em> Organoid Culture of Primary Mouse Colon<span style='display: none;'> Tumors</span>…

In vitro Organoid Culture of Primary Mouse Colon Tumors…
Published 5/17/2013

A simple method to establish primary murine colon tumor organoid is described. This method utilizes the feature that colon tumor cells survive and grow into organoids in media containing limited growth factors, whereas normal colon epithelial do not.

Mouse Models for Graft Arteriosclerosis

Mouse Models for Graft Arteriosclerosis
Published 5/14/2013

We describe protocols for our mouse graft arteriosclerois (GA) models which involve interposition of a mouse vessel segment into a recipient of the same inbred strain. By backcrossing additional genetic changes into the vessel donor, the model can assess the effect of specific genes on GA.

Air Filter Devices Including Nonwoven Meshes of Electrospun <span style='display: none;'>Recombinant Spider Silk Proteins</span>…

Air Filter Devices Including Nonwoven Meshes of Electrospun Recombinant Spider Silk Proteins…
Published 5/08/2013

Spider silk fibers display extraordinary mechanical properties. Engineered Araneus diadematus Fibroin 4 (eADF4) can be processed into nonwoven meshes using electrospinning. Here, the eADF4 nonwoven meshes are used to improve the performance of air filtering devices.

Biosensor for Detection of Antibiotic Resistant<span style='display: none;'> Staphylococcus Bacteria</span>…

Biosensor for Detection of Antibiotic Resistant Staphylococcus Bacteria…
Published 5/08/2013

Lytic phage biosensors and antibody beads are able to discriminate between methicillin resistant (MRSA) and sensitive staphylococcus bacteria. The phages were immobilized by a Langmuir-Blodgett method onto a surface of a quartz crystal microbalance sensor and worked as broad range staphylococcus probes. Antibody beads recognize MRSA.

Manufacturing Devices and Instruments for Easier Rat Liver<span style='display: none;'> Transplantation</span>…

Manufacturing Devices and Instruments for Easier Rat Liver Transplantation…
Published 5/05/2013

We describe the design of the “quick-linker” device for easier orthotopic rat liver transplantation.

Microfluidic-based Electrotaxis for On-demand Quantitative<span style='display: none;'> Analysis of Caenorhabditis elegans' Locomotion</span>…

Microfluidic-based Electrotaxis for On-demand Quantitative…
Published 5/02/2013

A semi-automated micro-electro-fluidic method to induce on-demand locomotion in Caenorhabditis elegans is described. This method is based on the neurophysiologic phenomenon of worms responding to mild electric fields (“electrotaxis”) inside microfluidic channels. Microfluidic electrotaxis serves as a rapid, sensitive, low-cost, and scalable technique to screen for factors affecting neuronal health.

Measuring Spatially- and Directionally-varying Light<span style='display: none;'> Scattering from Biological Material</span>…

Measuring Spatially- and Directionally-varying Light Scattering from Biological Material…
Published 5/20/2013

We present a non-destructive method for sampling spatial variation in the direction of light scattered from structurally complex materials. By keeping the material intact, we preserve gross-scale scattering behavior, while concurrently capturing fine-scale directional contributions with high-resolution imaging. Results are visualized in software at biologically-relevant positions and scales.

Synthesis and Functionalization of Nitrogen-doped Carbon<span style='display: none;'> Nanotube Cups with Gold Nanoparticles as Cork Stoppers</span>…

Synthesis and Functionalization of Nitrogen-doped Carbon…
Published 5/13/2013

We discussed the synthesis of individual graphitic nanocups using a series of techniques including chemical vapor deposition, acid oxidation and probe-tip sonication. By citrate reduction of HAuCl₄, the graphitic nanocups were effectively corked with gold nanoparticles due to the chemically reactive edges of the cups.

Construction of a High Resolution Microscope with<span style='display: none;'> Conventional and Holographic Optical Trapping Capabilities</span>…

Construction of a High Resolution Microscope with…
Published 4/22/2013

The system described herein employs a traditional optical trap as well as an independent holographic optical trapping line, capable of creating and manipulating multiple traps. This allows for the creation of complex geometric arrangements of refractive particles while also permitting simultaneous high-speed, high-resolution measurements of the activity of biological enzymes.

Atom Probe Tomography Studies on the<span style='display: none;'> Cu(In,Ga)Se2 Grain Boundaries</span>…

Atom Probe Tomography Studies on the Cu(In,Ga)Se2 Grain Boundaries…
Published 4/22/2013

In this work, we describe the use of the atom-probe tomography technique for studying the grain boundaries of the absorber layer in a CIGS solar cell. A novel approach to prepare the atom probe tips containing the desired grain boundary with a known structure is also presented here.

Untargeted Metabolomics from Biological Sources Using<span style='display: none;'> Ultraperformance Liquid Chromatography-High Resolution Mass Spectrometry (UPLC-HRMS)</span>…

Untargeted Metabolomics from Biological Sources Using…
Published 5/20/2013

Untargeted metabolomics provides a hypothesis generating snapshot of a metabolic profile. This protocol will demonstrate the extraction and analysis of metabolites from cells, serum, or tissue. A range of metabolites are surveyed using liquid-liquid phase extraction, microflow ultraperformance liquid chromatography/high-resolution mass spectrometry (UPLC-HRMS) coupled to differential analysis software.

Fabricating Nanogaps by Nanoskiving

Fabricating Nanogaps by Nanoskiving
Published 5/13/2013

The fabrication of electrically addressable, high-aspect-ratio (> 1000:1) metal nanowires separated by gaps of single nanometers using either sacrificial layers of aluminum and silver or self-assembled monolayers as templates is described. These nanogap structures are fabricated without a clean room or any photo- or electron-beam lithographic processes by a form of edge lithography known as nanoskiving.

Free Radicals in Chemical Biology: from Chemical Behavior<span style='display: none;'> to Biomarker Development</span>…

Free Radicals in Chemical Biology: from Chemical Behavior to Biomarker Development…
Published 4/15/2013

Radical-based biomimetic chemistry has been applied to building-up libraries necessary for biomarker development.

Microwave-assisted Intramolecular Dehydrogenative<span style='display: none;'> Diels-Alder Reactions for the Synthesis of Functionalized Naphthalenes/Solvatochromic Dyes</span>…

Microwave-assisted Intramolecular Dehydrogenative…
Published 4/01/2013

Microwave-assisted intramolecular dehydrogenative Diels-Alder (DA) reactions provide concise access to functionalized cyclopenta[b]naphthalene building blocks. The utility of this methodology is demonstrated by one-step conversion of the dehydrogenative DA cycloadducts into novel solvatochromic fluorescent dyes via Buchwald-Hartwig palladium-catalyzed cross-coupling reactions.

JoVE Behavior (opening in June, 2013) includes both observational and experimental techniques that seek to understand human and animal behavior, implicated brain regions, related physiological changes, and underlying genetic causes. The scope of section includes but is not limited to:

Cognition (Attention, Decision Making, Reasoning)
Sexual and Motivational Behaviors
Social Behaviors
Learning and Memory
Sleep and biological rhythms
Language Processing
Addiction
Emotion
Control of Movement
Consciousness

Call for Papers: Send inquiries to behavior@jove.com

JoVE Environment (opening in September, 2013) is a multidisciplinary section devoted to the publication of research methods employed in environmental science and green technologies, from biofuels to oceanography to atmospheric sciences. Special consideration is given to experimental techniques and approaches aimed at understanding the Earth, protecting natural resources and creating a more sustainable and environmentally-conscious planet. The scope of this section includes but is not limited to:

Renewable energy and sustainable materials
Environmental engineering
Basic and applied ecology
Ecotoxicology and ecological health
Marine biology and oceanography
Soil and agricultural sciences
Forestry and botany
Atmospheric science and climate modeling
Geosciences

Call for Papers: Send inquiries to environment@jove.com

Most Popular Video Articles

Hi-C: A Method to Study the Three-dimensional Architecture<span style='display: none;'> of Genomes.</span>…

Hi-C: A Method to Study the Three-dimensional Architecture of Genomes.…
Published 5/06/2010

The Hi-C method allows unbiased, genome-wide identification of chromatin interactions (1). Hi-C couples proximity ligation and massively parallel sequencing. The resulting data can be used to study genomic architecture at multiple scales: initial results identified features such as chromosome territories, segregation of open and closed chromatin, and chromatin structure at the megabase scale.

Microvolume Protein Concentration Determination using the<span style='display: none;'> NanoDrop 2000c Spectrophotometer</span>…

Microvolume Protein Concentration Determination using the NanoDrop 2000c Spectrophotometer…
Published 11/04/2009

Microvolume samples are quantified by a spectrophotometer system that uses natural surface tension to retain samples without the use of cuvettes or capillaries. The dynamic range of protein concentrations and speed by which they can be measured are greatly increased with this method.

Isolation of Mouse Peritoneal Cavity Cells

Isolation of Mouse Peritoneal Cavity Cells
Published 1/28/2010

The peritoneal cavity in mammals contains different immune cell populations crucial for innate immune responses. An efficient isolation method is required for biochemical and functional analyses of these cells. Here we provide a comprehensive method for the isolation of peritoneal cavity cells in the mouse.

Generating iPS Cells from MEFS through Forced Expression of <span style='display: none;'>Sox-2, Oct-4, c-Myc, and Klf4</span>…

Generating iPS Cells from MEFS through Forced Expression of Sox-2, Oct-4, c-Myc, and Klf4…
Published 4/07/2008

This video shows the procedure for generating induced pluripotent stem cells using inducible lentivirus that express Oct4, Sox2, c-Myc and Klf4.

Hi-C: A Method to Study the Three-dimensional Architecture<span style='display: none;'> of Genomes.</span>…

Hi-C: A Method to Study the Three-dimensional Architecture of Genomes.…
Published 5/06/2010

The Hi-C method allows unbiased, genome-wide identification of chromatin interactions (1). Hi-C couples proximity ligation and massively parallel sequencing. The resulting data can be used to study genomic architecture at multiple scales: initial results identified features such as chromosome territories, segregation of open and closed chromatin, and chromatin structure at the megabase scale.

Microvolume Protein Concentration Determination using the<span style='display: none;'> NanoDrop 2000c Spectrophotometer</span>…

Microvolume Protein Concentration Determination using the NanoDrop 2000c Spectrophotometer…
Published 11/04/2009

Microvolume samples are quantified by a spectrophotometer system that uses natural surface tension to retain samples without the use of cuvettes or capillaries. The dynamic range of protein concentrations and speed by which they can be measured are greatly increased with this method.

Generating iPS Cells from MEFS through Forced Expression of <span style='display: none;'>Sox-2, Oct-4, c-Myc, and Klf4</span>…

Generating iPS Cells from MEFS through Forced Expression of Sox-2, Oct-4, c-Myc, and Klf4…
Published 4/07/2008

This video shows the procedure for generating induced pluripotent stem cells using inducible lentivirus that express Oct4, Sox2, c-Myc and Klf4.

Dissection of Hippocampal Dentate Gyrus from Adult Mouse

Dissection of Hippocampal Dentate Gyrus from Adult Mouse
Published 11/17/2009

A dissection technique for removal of the dentate gyrus from adult mouse under a stereomicroscope was demonstrated in this video-recorded protocol.

Electrode Positioning and Montage in Transcranial Direct<span style='display: none;'> Current Stimulation</span>…

Electrode Positioning and Montage in Transcranial Direct Current Stimulation…
Published 5/23/2011

Transcranial direct current stimulation (tDCS) is an established technique to modulate cortical excitability^1,2. It has been used as an investigative tool in neuroscience due to its effects on cortical plasticity, easy operation, and safe profile. One area that tDCS has been showing encouraging results is pain alleviation ^3-5.

Focal Cerebral Ischemia Model by Endovascular Suture<span style='display: none;'> Occlusion of the Middle Cerebral Artery in the Rat</span>…

Focal Cerebral Ischemia Model by Endovascular Suture…
Published 2/05/2011

Surgical induction of ischemic brain damage in the rat is a widely used model for stroke research. Here we demonstrate the induction of focal cerebral ischemia by occlusion of the middle cerebral artery. Visualization of the resulting infarct by histological staining and magnetic resonance imaging is also shown.

Preparation of Acute Hippocampal Slices from Rats and<span style='display: none;'> Transgenic Mice for the Study of Synaptic Alterations during Aging and Amyloid Pathology</span>…

Preparation of Acute Hippocampal Slices from Rats and…
Published 3/23/2011

This article outlines procedures for preparing hippocampal slices from rats and transgenic mice for the study of synaptic alterations associated with brain aging and age-related neurodegenerative diseases, such as Alzheimer’s disease.

The Subventricular Zone En-face: Wholemount Staining and<span style='display: none;'> Ependymal Flow</span>…

The Subventricular Zone En-face: Wholemount Staining and Ependymal Flow…
Published 5/06/2010

The lateral ventricle walls contain the largest germinal region in the adult mammalian brain. Traditionally, studies on neurogenesis in this region have relied on classical sectioning techniques for histological analysis. Here we present an alternative approach, the wholemount technique, which provides a comprehensive, en-face view of this germinal region.

Isolation of Mouse Peritoneal Cavity Cells

Isolation of Mouse Peritoneal Cavity Cells
Published 1/28/2010

The peritoneal cavity in mammals contains different immune cell populations crucial for innate immune responses. An efficient isolation method is required for biochemical and functional analyses of these cells. Here we provide a comprehensive method for the isolation of peritoneal cavity cells in the mouse.

Neutrophil Extracellular Traps: How to Generate and<span style='display: none;'> Visualize Them</span>…

Neutrophil Extracellular Traps: How to Generate and Visualize Them…
Published 2/24/2010

Neutrophil Extracellular Traps (NETs) are an important innate immune mechanism to fight pathogenic bacteria, fungi and parasites. Here we describe methods to isolate neutrophil granulocytes from human blood and to activate them to form NETs. We present preparation techniques to visualize NETs in light and electron microscopy.

Determination of Molecular Structures of HIV Envelope<span style='display: none;'> Glycoproteins using Cryo-Electron Tomography and Automated Sub-tomogram Averaging</span>…

Determination of Molecular Structures of HIV Envelope…
Published 12/01/2011

The protocol describes a high-throughput approach to determining structures of membrane proteins using cryo-electron tomography and 3D image processing. It covers the details of specimen preparation, data collection, data processing and interpretation, and concludes with the production of a representative target for the approach, the HIV-1 Envelope glycoprotein. These computational procedures are designed in a way that enables researchers and students to work remotely and contribute to data processing and structural analysis.

IgY Technology: Extraction of Chicken Antibodies from Egg<span style='display: none;'> Yolk by Polyethylene Glycol (PEG) Precipitation</span>…

IgY Technology: Extraction of Chicken Antibodies from Egg…
Published 5/01/2011

This protocol describes in particular the extraction of total IgY from egg yolk by means of polyethylene glycol precipitation and gives general information about IgY technology.

Reverse Total Shoulder Arthroplasty

Reverse Total Shoulder Arthroplasty
Published 7/05/2011

Reverse total shoulder arthroplasty is indicated for the treatment of conditions that cannot be treated with conventional arthroplasty or other procedures. These primarily include degenerative and incapacitating conditions with irreparable rotator cuff and loss of the normal biomechanical coupling of the shoulder.

Evaluation of Cancer Stem Cell Migration Using<span style='display: none;'> Compartmentalizing Microfluidic Devices and Live Cell Imaging</span>…

Evaluation of Cancer Stem Cell Migration Using…
Published 12/23/2011

A compartmentalizing microfluidic device for investigating cancer stem cell migration is described. This novel platform creates a viable cellular microenvironment and enables microscopic visualization of live cell locomotion. Highly motile cancer cells are isolated to study molecular mechanisms of aggressive infiltration, potentially leading to more effective future therapies.

Mouse Model of Middle Cerebral Artery Occlusion

Mouse Model of Middle Cerebral Artery Occlusion
Published 2/13/2011

We demonstrate in the video a method for producing a middle cerebral artery occlusion in adult mice using an intraluminal monofilament. We also show how to evaluate the extent of cerebral infarction by 2,3,5-triphenyltetrazolium chloride (TTC) staining.

Coronary Artery Ligation and Intramyocardial Injection in a <span style='display: none;'>Murine Model of Infarction</span>…

Coronary Artery Ligation and Intramyocardial Injection in a Murine Model of Infarction…
Published 6/07/2011

Numerous genetic manipulations and/or intramyocardial injections of genes, proteins, cells, and/or biomaterials are superimposed upon the dimension of time in studies of acute ischemia/ reperfusion injury and chronic remodeling in mice. This video illustrates the microsurgical procedures for ischemia/reperfusion, permanent coronary artery ligation, and intramyocardial injection studies.

Creating Transient Cell Membrane Pores Using a Standard<span style='display: none;'> Inkjet Printer</span>…

Creating Transient Cell Membrane Pores Using a Standard Inkjet Printer…
Published 3/16/2012

A description of the methods used to convert an HP DeskJet 500 printer into a bioprinter. The printer is capable of processing living cells, which causes transient pores in the membrane. These pores can be utilized to incorporate small molecules, including fluorescent G-actin, into the printed cells.

Design of a Cyclic Pressure Bioreactor for the <em>Ex</em><span style='display: none;'> Vivo Study of Aortic Heart Valves</span>…

Design of a Cyclic Pressure Bioreactor for the Ex Vivo Study of Aortic Heart Valves…
Published 8/23/2011

A cyclic pressure bioreactor capable of subjecting heart valve tissue to physiological and pathological pressure conditions has been designed. A LabVIEW program allows users to control pressure magnitude, amplitude and frequency. This device can be used to study the mechanobiology of heart valve tissue or isolated cells.

Procedure for Lung Engineering

Procedure for Lung Engineering
Published 3/08/2011

We have developed a decellularized lung extracellular matrix and novel biomimetic bioreactor that can be used to generate functional lung tissue. By seeding cells into the matrix and culturing in the bioreactor, we generate tissue that demonstrates effective gas exchange when transplanted in vivo for short periods of time.

Formulation of Diblock Polymeric Nanoparticles through<span style='display: none;'> Nanoprecipitation Technique</span>…

Formulation of Diblock Polymeric Nanoparticles through Nanoprecipitation Technique…
Published 9/20/2011

This article describes a nanoprecipitation method to synthesize polymer-based nanoparticles using diblock co-polymers. We will discuss the synthesis of diblock co-polymers, the nanoprecipitation technique, and potential applications.

Polycrystalline Silicon Thin-film Solar cells with<span style='display: none;'> Plasmonic-enhanced Light-trapping</span>…

Polycrystalline Silicon Thin-film Solar cells with Plasmonic-enhanced Light-trapping…
Published 7/02/2012

Polycrystalline silicon thin-film solar cells on glass are fabricated by deposition of boron and phosphorous doped silicon layers followed by crystallisation, defect passivation and metallisation. Plasmonic light-trapping is introduced by forming Ag nanoparticles on the silicon cell surface capped with a diffused reflector resulting in ~45% photocurrent enhancement.

Investigation of Early Plasma Evolution Induced by<span style='display: none;'> Ultrashort Laser Pulses</span>…

Investigation of Early Plasma Evolution Induced by Ultrashort Laser Pulses…
Published 7/02/2012

An experimental method to examine the early plasma evolution induced by ultrashort laser pulses is described. Using this method, high quality images of early plasma are obtained with high temporal and spatial resolutions. A novel integrated atomistic model is used to simulate and explain the mechanisms of early plasma.

Fabrication, Densification, and Replica Molding of 3D<span style='display: none;'> Carbon Nanotube Microstructures</span>…

Fabrication, Densification, and Replica Molding of 3D Carbon Nanotube Microstructures…
Published 7/02/2012

We present methods for fabrication of patterned microstructures of vertically aligned carbon nanotubes (CNTs), and their use as master molds for production of polymer microstructures with organized nanoscale surface texture. The CNT forests are densified by condensation of solvent onto the substrate, which significantly increases their packing density and enables self-directed formation of 3D shapes.

Controlling the Size, Shape and Stability of Supramolecular <span style='display: none;'>Polymers in Water</span>…

Controlling the Size, Shape and Stability of Supramolecular Polymers in Water…
Published 8/02/2012

The goal of this experiment is to determine and control the size, shape and stability of self-assembled discotic amphiphiles in water. For aqueous based supramolecular polymers such level of control is very difficult. We apply a strategy using both repulsive and attractive interactions. The experimental techniques applied to characterize this system are broadly applicable.

Rapid Colorimetric Assays to Qualitatively Distinguish RNA<span style='display: none;'> and DNA in Biomolecular Samples</span>…

Rapid Colorimetric Assays to Qualitatively Distinguish RNA and DNA in Biomolecular Samples…
Published 2/04/2013

A suite of colorimetric assays is described for rapidly distinguishing protein, RNA, DNA, and reducing sugars in potentially heterogeneous biomolecular samples.

Production of Disulfide-stabilized Transmembrane Peptide<span style='display: none;'> Complexes for Structural Studies</span>…

Production of Disulfide-stabilized Transmembrane Peptide Complexes for Structural Studies…
Published 3/06/2013

Biophysical and biochemical studies of interactions among membrane-embedded protein domains face many technical challenges, the first of which is obtaining appropriate study material. This article describes a protocol for producing and purifying disulfide-stabilized transmembrane peptide complexes that are suitable for structural analysis by solution nuclear magnetic resonance (NMR) and other analytical applications.

Origami Inspired Self-assembly of Patterned and<span style='display: none;'> Reconfigurable Particles</span>…

Origami Inspired Self-assembly of Patterned and Reconfigurable Particles…
Published 2/04/2013

We describe experimental details of the synthesis of patterned and reconfigurable particles from two dimensional (2D) precursors. This methodology can be used to create particles in a variety of shapes including polyhedra and grasping devices at length scales ranging from the micro to centimeter scale.

Preparation and Use of Samarium Diiodide (SmI<sub>2</sub>)<span style='display: none;'> in Organic Synthesis: The Mechanistic Role of HMPA and Ni(II) Salts in the Samarium Barbier Reaction</span>…

Preparation and Use of Samarium Diiodide (SmI₂)…
Published 2/04/2013

A straightforward procedure for the preparation of samarium diiodide (SmI₂) in THF is described. The role of two main additives namely hexamethylphosphoramide (HMPA) and Ni(acac)₂ in Sm mediated reactions is demonstrated in the Sm-Barbier reaction.

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