8,715 results match your criteria Journal of Visualized Experiments [Journal]


Erratum: Comparable Decellularization of Fetal and Adult Cardiac Tissue Explants as 3D-like Platforms for In Vitro Studies.

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J Vis Exp 2019 Apr 23(146). Epub 2019 Apr 23.

An erratum was issued for: Comparable Decellularization of Fetal and Adult Cardiac Tissue Explants as 3D-like Platforms for In Vitro Studies.  The affiliations were updated. The fourth affiliation was corrected from: Gladstone Institutes, University of California San Francisco to: Gladstone Institute of Cardiovascular Disease An additional affiliation was added for Todd C. Read More

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http://dx.doi.org/10.3791/6318DOI Listing
April 2019
1 Read

Capturing Small Molecule Communication Between Tissues and Cells Using Imaging Mass Spectrometry.

J Vis Exp 2019 Apr 3(146). Epub 2019 Apr 3.

Department of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago;

Imaging mass spectrometry (IMS) has routinely been applied to three types of samples: tissue sections, spheroids, and microbial colonies. These sample types have been analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to visualize the distribution of proteins, lipids, and metabolites across the biological sample of interest. We have developed a novel sample preparation method that combines the strengths of the three previous applications to address an underexplored approach for identifying chemical communication in cancer, by seeding mammalian cell cultures into agarose in coculture with healthy tissues followed by desiccation of the sample. Read More

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http://dx.doi.org/10.3791/59490DOI Listing

Ion Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization.

J Vis Exp 2019 Apr 5(146). Epub 2019 Apr 5.

Wolfson Centre for Applied Structural Biology, The Alexander Silberman Institute of Life Science, The Hebrew University of Jerusalem;

Ion-exchange chromatography with multi-angle light scattering (IEX-MALS) is a powerful method for protein separation and characterization. The combination of the high-specificity separation technique IEX with the accurate molar mass analysis achieved by MALS allows the characterization of heterogeneous protein samples, including mixtures of oligomeric forms or protein populations, even with very similar molar masses. Therefore, IEX-MALS provides an additional level of protein characterization and is complementary to the standard size-exclusion chromatography with multi-angle light scattering (SEC-MALS) technique. Read More

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http://dx.doi.org/10.3791/59408DOI Listing

Opsonophagocytic Killing Assay to Assess Immunological Responses Against Bacterial Pathogens.

J Vis Exp 2019 Apr 5(146). Epub 2019 Apr 5.

Department of Biochemistry and Molecular Biology, Center for Molecular Medicine and Complex Carbohydrate Research Center, University of Georgia;

A key aspect of the immune response to bacterial colonization of the host is phagocytosis. An opsonophagocytic killing assay (OPKA) is an experimental procedure in which phagocytic cells are co-cultured with bacterial units. The immune cells will phagocytose and kill the bacterial cultures in a complement-dependent manner. Read More

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http://dx.doi.org/10.3791/59400DOI Listing

Purification of Extracellular Trypanosomes, Including African, from Blood by Anion-Exchangers (Diethylaminoethyl-cellulose Columns).

J Vis Exp 2019 Apr 6(146). Epub 2019 Apr 6.

Univ. Bordeaux UMR INTERTRYP 177 IRD CIRAD; IRD UMR 177 INTERTRYP CIRAD; Centre Hospitalier Université Bordeaux;

This method allows the separation of trypanosomes, parasites responsible for animal and human African trypanosomiasis (HAT), from infected blood. This is the best method for diagnosis of first stage HAT and furthermore this parasite purification method permits serological and research investigations. HAT is caused by Tsetse fly transmitted Trypanosoma brucei gambiense and T. Read More

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http://dx.doi.org/10.3791/58415DOI Listing

A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging.

J Vis Exp 2019 Apr 8(146). Epub 2019 Apr 8.

CREOL, The College of Optics and Photonics, University of Central Florida;

Single-molecule imaging has greatly advanced our understanding of molecular mechanisms in biological studies. However, it has been challenging to obtain large field-of-view, high-contrast images in thick cells and tissues. Here, we introduce highly inclined swept tile (HIST) microscopy that overcomes this problem. Read More

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https://www.jove.com/video/59360/a-guide-to-build-highly-inc
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http://dx.doi.org/10.3791/59360DOI Listing
April 2019
6 Reads

Swimming Induced Paralysis to Assess Dopamine Signaling in Caenorhabditis elegans.

J Vis Exp 2019 Apr 3(146). Epub 2019 Apr 3.

Harriet Wilkes Honors College, Florida Atlantic University, John D MacArthur Campus; Brain Institute, Florida Atlantic University; Department of Biomedical Science, Charles E. Schmidt College of Medicine, Florida Atlantic University;

The swimming assay described in this protocol is a valid tool to identify proteins regulating the dopaminergic synapses. Similar to mammals, dopamine (DA) controls several functions in C. elegans including learning and motor activity. Read More

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http://dx.doi.org/10.3791/59243DOI Listing

Remote Laboratory Management: Respiratory Virus Diagnostics.

J Vis Exp 2019 Apr 6(146). Epub 2019 Apr 6.

Baylor Global Health, Baylor College of Medicine; Department of Medicine, Baylor College of Medicine;

An uptick in recent pandemics (Ebola, Zika, MERS, influenza, etc.) underlines the need for a more 'nimble,' coordinated response that addresses a multitude of issues ranging from transportation, access, facilities, equipment, and communication to provider training. To address this need, we have developed an innovative, scalable, logistics-enhanced, mobile, laboratory facility for emergencies and epidemics in resource-constrained global settings. Read More

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http://dx.doi.org/10.3791/59188DOI Listing

Behavioral Tracking and Neuromast Imaging of Mexican Cavefish.

J Vis Exp 2019 Apr 6(146). Epub 2019 Apr 6.

Department of Biology, University of Hawai'i at Mānoa;

Cave-dwelling animals have evolved a series of morphological and behavioral traits to adapt to their perpetually dark and food-sparse environments. Among these traits, foraging behavior is one of the useful windows into functional advantages of behavioral trait evolution. Presented herein are updated methods for analyzing vibration attraction behavior (VAB: an adaptive foraging behavior) and imaging of associated mechanosensors of cave-adapted tetra, Astyanax mexicanus. Read More

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http://dx.doi.org/10.3791/59099DOI Listing

Generation of Monoclonal Antibodies Against Natural Products.

J Vis Exp 2019 Apr 6(146). Epub 2019 Apr 6.

Center of Scientific Experiment, Beijing University of Chinese Medicine;

The analysis of the bioactive components present in foods and natural products has become a popular area of study in many fields, including traditional Chinese medicine and food safety/toxicology. Many of the classical analysis techniques require expensive equipment and/or expertise. Notably, enzyme-linked immunosorbent assays (ELISAs) have become an emerging method for the analysis of foods and natural products. Read More

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http://dx.doi.org/10.3791/57116DOI Listing

Human Serum Anti-aquaporin-4 Immunoglobulin G Detection by Cell-based Assay.

J Vis Exp 2019 Apr 5(146). Epub 2019 Apr 5.

Department of Neurology and Neuroscience Center, The First Hospital of Jilin University;

Anti-aquaporin-4 (AQP4) immunoglobulin G (IgG) is the core diagnostic biomarker for neuromyelitis optica spectrum disorders (NMOSD). The cell-based assay (CBA) is a widely used method to detect anti-AQP4 IgG in human serum with high sensitivity and specificity. Briefly, serum anti-AQP4 IgG is captured by AQP4-transfected cell that is fixed on the biochip then detected by a fluorescein-labelled secondary antibody. Read More

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https://www.jove.com/video/59014/human-serum-anti-aquaporin-
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http://dx.doi.org/10.3791/59014DOI Listing
April 2019
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Multi-Modal Signals for Analyzing Pain Responses to Thermal and Electrical Stimuli.

J Vis Exp 2019 Apr 5(146). Epub 2019 Apr 5.

Department of Medical Psychology, Ulm University.

The assessment of pain relies mostly on methods that require a person to communicate. However, for people with cognitive and verbal impairments, existing methods are not sufficient as they lack reliability and validity. To approach this problem, recent research focuses on an objective pain assessment facilitated by parameters of responses derived from physiology, and video and audio signals. Read More

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http://dx.doi.org/10.3791/59057DOI Listing

Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses.

J Vis Exp 2019 Apr 4(146). Epub 2019 Apr 4.

UMR 1173 Institut national de la santé et de la recherche médicale (INSERM), Université de Versailles St. Quentin; Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpital Ambroise Paré, Laboratoire de Microbiologie;

The use of recombinant viruses has become crucial in basic or applied virology. Reverse genetics has been proven to be an extremely powerful technology, both to decipher viral replication mechanisms and to study antivirals or provide development platform for vaccines. The construction and manipulation of a reverse genetic system for a negative-strand RNA virus such as a respiratory syncytial virus (RSV), however, remains delicate and requires special know-how. Read More

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http://dx.doi.org/10.3791/59218DOI Listing

Long-term Video Tracking of Cohoused Aquatic Animals: A Case Study of the Daily Locomotor Activity of the Norway Lobster (Nephrops norvegicus).

J Vis Exp 2019 Apr 8(146). Epub 2019 Apr 8.

Institute of Marine Sciences, Spanish National Research Council (CSIC).

We present a protocol related to a video-tracking technique based on the background subtraction and image thresholding that makes it possible to individually track cohoused animals. We tested the tracking routine with four cohoused Norway lobsters (Nephrops norvegicus) under light-darkness conditions for 5 days. The lobsters had been individually tagged. Read More

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http://dx.doi.org/10.3791/58515DOI Listing

Real-time Live-cell Flow Cytometry to Investigate Calcium Influx, Pore Formation, and Phagocytosis by P2X7 Receptors in Adult Neural Progenitor Cells.

J Vis Exp 2019 Apr 3(146). Epub 2019 Apr 3.

Florey Institute of Neuroscience and Mental Health, University of Melbourne;

Live-cell flow cytometry is increasingly used among cell biologists to quantify biological processes in a living cell culture. This protocol describes a method whereby live-cell flow cytometry is extended upon to analyze the multiple functions of P2X7 receptor activation in real-time. Using a time module installed on a flow cytometer, live-cell functionality can be assessed and plotted over a given time period to explore the kinetics of calcium influx, transmembrane pore formation, and phagocytosis. Read More

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http://dx.doi.org/10.3791/59313DOI Listing

Mimicking a Space Mission to Mars Using Hindlimb Unloading and Partial Weight Bearing in Rats.

J Vis Exp 2019 Apr 4(146). Epub 2019 Apr 4.

Department of Neurology, Harvard Medical School - Beth Israel Deaconess Medical Center.

Rodent ground-based models are widely used to understand the physiological consequences of space flight on the physiological system and have been routinely employed since 1979 and the development of hind limb unloading (HLU). However, the next steps in space exploration now include to travel to Mars where the gravity is 38% of Earth's gravity. Since no human being has experienced this level of partial gravity, a sustainable ground-based model is necessary to investigate how the body, already impaired by the time spent in microgravity, would react to this partial load. Read More

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http://dx.doi.org/10.3791/59327DOI Listing

Studying Brain Function in Children Using Magnetoencephalography.

J Vis Exp 2019 Apr 8(146). Epub 2019 Apr 8.

ARC Centre of Excellence in Cognition and its Disorders, Macquarie University; Department of Cognitive Science, Macquarie University;

Magnetoencephalography (MEG) is a non-invasive neuroimaging technique which directly measures magnetic fields produced by the electrical activity of the human brain. MEG is quiet and less likely to induce claustrophobia compared with magnetic resonance imaging (MRI). It is therefore a promising tool for investigating brain function in young children. Read More

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http://dx.doi.org/10.3791/58909DOI Listing

An Instrumented Pull Test to Characterize Postural Responses.

J Vis Exp 2019 Apr 6(146). Epub 2019 Apr 6.

Department of Medical Bionics, The University of Melbourne; Department of Neurology, Austin Hospital;

Impairment of postural reflexes, termed postural instability, is a common and disabling deficit in Parkinson's disease. To assess postural reflexes, clinicians typically employ the pull test to grade corrective responses to a backward perturbation at the shoulders. However, the pull test is prone to issues with reliability and scaling (score/4). Read More

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http://dx.doi.org/10.3791/59309DOI Listing

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression.

J Vis Exp 2019 Mar 29(145). Epub 2019 Mar 29.

Center for Epigenetics, Van Andel Research Institute;

Here we describe a protocol for implementing the REMOTE-control system (Reversible Manipulation of Transcription at Endogenous loci), which allows for reversible and tunable expression control of an endogenous gene of interest in living model systems. The REMOTE-control system employs enhanced lac repression and tet activation systems to achieve down- or upregulation of a target gene within a single biological system. Tight repression can be achieved from repressor binding sites flexibly located far downstream of a transcription start site by inhibiting transcription elongation. Read More

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http://dx.doi.org/10.3791/59235DOI Listing

Effects of Blast-induced Neurotrauma on Pressurized Rodent Middle Cerebral Arteries.

J Vis Exp 2019 Apr 1(146). Epub 2019 Apr 1.

Charles R. Allen Research Laboratories, Department of Anesthesiology, University of Texas Medical Branch; The Moody Project for Translational Traumatic Brain Injury Research, University of Texas Medical Branch.

Though there have been studies on the histopathological and behavioral effects of blast exposure, fewer have been dedicated to blast's cerebral vascular effects. Impact (i.e. Read More

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http://dx.doi.org/10.3791/58792DOI Listing

HOX Loci Focused CRISPR/sgRNA Library Screening Identifying Critical CTCF Boundaries.

J Vis Exp 2019 Mar 31(145). Epub 2019 Mar 31.

Department of Pediatrics, Pennsylvania State University College of Medicine;

CCCTC-binding factor (CTCF)-mediated stable topologically associating domains (TADs) play a critical role in constraining interactions of DNA elements that are located in neighboring TADs. CTCF plays an important role in regulating the spatial and temporal expression of HOX genes that control embryonic development, body patterning, hematopoiesis, and leukemogenesis. However, it remains largely unknown whether and how HOX loci associated CTCF boundaries regulate chromatin organization and HOX gene expression. Read More

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https://www.jove.com/video/59382/hox-loci-focused-crisprsgrn
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http://dx.doi.org/10.3791/59382DOI Listing
March 2019
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Determining the Egg Fertilization Rate of Bemisia tabaci Using a Cytogenetic Technique.

J Vis Exp 2019 Apr 1(146). Epub 2019 Apr 1.

Department of Entomology, University of Arizona.

A few species of sap-sucking whiteflies are some of the most damaging terrestrial pests worldwide because of the crop damage they inflict and plant viruses they vector. Despite numerous studies of the biology of these species in different environments, a key life history parameter, offspring sex ratios, has received little attention, yet is important for predicting population dynamics. The primary sex ratio (sex ratio at oviposition) of Bemisia tabaci has never been reported but can be found by determining the egg fertilization rate of this haplodiploid insect. Read More

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https://www.jove.com/video/59213/determining-egg-fertilizati
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http://dx.doi.org/10.3791/59213DOI Listing
April 2019
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DNA Electrophoresis Using Thiazole Orange Instead of Ethidium Bromide or Alternative Dyes.

J Vis Exp 2019 Mar 31(145). Epub 2019 Mar 31.

Department of Molecular Biology and Chemistry, Christopher Newport University;

DNA gel electrophoresis using agarose is a common tool in molecular biology laboratories, allowing separation of DNA fragments by size. After separation, DNA is visualized by staining. This article demonstrates how to use thiazole orange to stain DNA. Read More

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https://www.jove.com/video/59341/dna-electrophoresis-using-t
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http://dx.doi.org/10.3791/59341DOI Listing
March 2019
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Driving Under the Influence: How Music Listening Affects Driving Behaviors.

J Vis Exp 2019 Mar 27(145). Epub 2019 Mar 27.

Laboratoire d'Etude des Mécanismes Cognitifs (EA 3082), University Lyon 2.

Car driving is a daily activity for many individuals in modern societies. Drivers often listen to music while driving. The method presented here investigates how listening to music influences driving behaviors. Read More

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https://www.jove.com/video/58342/driving-under-influence-how
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http://dx.doi.org/10.3791/58342DOI Listing
March 2019
2 Reads

Surface Properties of Synthesized Nanoporous Carbon and Silica Matrices.

J Vis Exp 2019 Mar 27(145). Epub 2019 Mar 27.

Department of Physicochemistry of Solid Surface, Faculty of Chemistry, Maria Curie-Skłodowska University.

In this work, we report the synthesis and characterization of ordered nanoporous carbon material (also called ordered mesoporous carbon material [OMC]) with a 4.6 nm pore size, and ordered silica porous matrix, SBA-15, with a 5.3 nm pore size. Read More

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https://www.jove.com/video/58395/surface-properties-synthesi
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http://dx.doi.org/10.3791/58395DOI Listing
March 2019
1 Read

Investigation of Plant Interactions Across Common Mycorrhizal Networks Using Rotated Cores.

J Vis Exp 2019 Mar 26(145). Epub 2019 Mar 26.

Department of Biology, University of Miami.

Arbuscular mycorrhizal (AM) fungi influence plant mineral nutrient uptake and growth, hence, they have the potential to influence plant interactions. The power of their influence is in extraradical mycelia that spread beyond nutrient depletion zones found near roots to ultimately interconnect individuals within a common mycorrhizal network (CMN). Most experiments, however, have investigated the role of AM fungi in plant interactions by growing plants with versus without mycorrhizal fungi, a method that fails to explicitly address the role of CMNs. Read More

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http://dx.doi.org/10.3791/59338DOI Listing

Magnetic Resonance Imaging Assessment of Carcinogen-induced Murine Bladder Tumors.

J Vis Exp 2019 Mar 29(145). Epub 2019 Mar 29.

Department of Urology, Northwestern University Feinberg School of Medicine;

Murine bladder tumor models are critical for the evaluation of new therapeutic options. Bladder tumors induced with the N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) carcinogen are advantageous over cell line-based models because they closely replicate the genomic profiles of human tumors, and, unlike cell models and xenografts, they provide a good opportunity for the study of immunotherapies. However, bladder tumor generation is heterogeneous; therefore, an accurate assessment of tumor burden is needed before randomization to experimental treatment. Read More

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https://www.jove.com/video/59101/magnetic-resonance-imaging-
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http://dx.doi.org/10.3791/59101DOI Listing
March 2019
1 Read

Lentiviral Vector Platform for the Efficient Delivery of Epigenome-editing Tools into Human Induced Pluripotent Stem Cell-derived Disease Models.

J Vis Exp 2019 Mar 29(145). Epub 2019 Mar 29.

Viral Vector Core, Duke University Medical Center;

The use of hiPSC-derived cells represents a valuable approach to study human neurodegenerative diseases. Here, we describe an optimized protocol for the differentiation of hiPSCs derived from a patient with the triplication of the alpha-synuclein gene (SNCA) locus into Parkinson's disease (PD)-relevant dopaminergic neuronal populations. Accumulating evidence has shown that high levels of SNCA are causative for the development of PD. Read More

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http://dx.doi.org/10.3791/59241DOI Listing
March 2019
1 Read

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT.

J Vis Exp 2019 Apr 1(146). Epub 2019 Apr 1.

Diabetes, Endocrinology, and Obesity Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health;

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT's ability to expend energy has made it a potential target for novel therapies to ameliorate obesity and associated metabolic disorders in humans. Though this tissue has been well studied in small animals, BAT's thermogenic capacity in humans remains largely unknown due to the difficulties of measuring its volume, activity, and distribution. Read More

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https://www.jove.com/video/58469/whole-body-regional-quantif
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http://dx.doi.org/10.3791/58469DOI Listing
April 2019
1 Read

Agrobacterium tumefaciens and Agrobacterium rhizogenes-Mediated Transformation of Potato and the Promoter Activity of a Suberin Gene by GUS Staining.

J Vis Exp 2019 Mar 29(145). Epub 2019 Mar 29.

Department of Biology, Universitat de Girona;

Agrobacterium sp. is one of the most widely used methods to obtain transgenic plants as it has the ability to transfer and integrate its own T-DNA into the plant's genome. Here, we present two transformation systems to genetically modify potato (Solanum tuberosum) plants. Read More

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http://dx.doi.org/10.3791/59119DOI Listing
March 2019
1 Read

A Method to Study the C924T Polymorphism of the Thromboxane A2 Receptor Gene.

J Vis Exp 2019 Apr 1(146). Epub 2019 Apr 1.

SS Annunziata University Hospital, Unit of Clinical Molecular Biology and Predictive Medicine, University of Chieti; Department of Medical, Oral and Biotechnological Sciences, University of Chieti.

The thromboxane A2 receptor (TBXA2R) gene is a member of the G-protein coupled superfamily with seven-transmembrane regions. It is involved in atherogenesis progression, ischemia, and myocardial infarction. Here we present a methodology of patient genotyping to investigate the post-transcriptional role of the C924T polymorphism (rs4523) situated at the 3' region of the TBXA2 receptor gene. Read More

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http://dx.doi.org/10.3791/57289DOI Listing
April 2019
1 Read

Seeding and Implantation of a Biosynthetic Tissue-engineered Tracheal Graft in a Mouse Model.

J Vis Exp 2019 Apr 1(146). Epub 2019 Apr 1.

Department of Otolaryngology Head & Neck Surgery, Nationwide Children's Hospital; Center for Regenerative Medicine, Research Institute at Nationwide Children's Hospital;

Treatment options for congenital or secondary long segment tracheal defects have historically been limited due to an inability to replace functional tissue. Tissue engineering holds great promise as a potential solution with its ability to integrate cells and signaling molecules into a 3-dimensional scaffold. Recent work with tissue engineered tracheal grafts (TETGs) has seen some success but their translation has been limited by graft stenosis, graft collapse, and delayed epithelialization. Read More

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https://www.jove.com/video/59173/seeding-implantation-biosyn
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http://dx.doi.org/10.3791/59173DOI Listing
April 2019
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Brachial Artery Catheterization in Swine.

J Vis Exp 2019 Mar 30(145). Epub 2019 Mar 30.

Neuroscience and Mental Health Institute, University of Alberta; Sensory Motor Adaptive Rehabilitation Technology (SMART) Network, University of Alberta; Division of Physical Medicine and Rehabilitation, University of Alberta.

The video describes in detail the catheterization of the distal brachial artery in swine. This technique enables researchers to measure arterial blood pressure continuously and collect arterial blood samples to assess arterial blood gas measurements. Arterial blood pressures and arterial blood gases are important physiological parameters to monitor during experimental procedures. Read More

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http://dx.doi.org/10.3791/59365DOI Listing
March 2019
1 Read

A Facile Synthetic Method to Obtain Bismuth Oxyiodide Microspheres Highly Functional for the Photocatalytic Processes of Water Depuration.

J Vis Exp 2019 Mar 29(145). Epub 2019 Mar 29.

Instituto de Ciencias Aplicadas y Tecnología, Universidad Nacional Autónoma de México.

Bismuth oxyhalide (BiOI) is a promising material for sunlight-driven-environmental photocatalysis. Given that the physical structure of this kind of materials is highly related to its photocatalytic performance, it is necessary to standardize the synthetic methods in order to obtain the most functional architectures and, thus, the highest photocatalytic efficiency. Here, we report a reliable route to obtain BiOI microspheres via the solvothermal process, using Bi(NO3)3 and potassium iodide (KI) as precursors, and ethylene glycol as a template. Read More

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http://dx.doi.org/10.3791/59006DOI Listing

Visualizing Impairment of the Endothelial and Glial Barriers of the Neurovascular Unit during Experimental Autoimmune Encephalomyelitis In Vivo.

J Vis Exp 2019 Mar 26(145). Epub 2019 Mar 26.

Theodor Kocher Institute, University of Bern;

The neurovascular unit (NVU) is composed of microvascular endothelial cells forming the blood-brain barrier (BBB), an endothelial basement membrane with embedded pericytes, and the glia limitans composed by the parenchymal basement membrane and astrocytic end-feed embracing the abluminal aspect of central nervous system (CNS) microvessels. In addition to maintaining CNS homeostasis the NVU controls immune cell trafficking into the CNS. During immunosurveillance of the CNS low numbers of activated lymphocytes can cross the endothelial barrier without causing BBB dysfunction or clinical disease. Read More

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http://dx.doi.org/10.3791/59249DOI Listing
March 2019
1 Read

Electroretinogram Recording in Larval Zebrafish using A Novel Cone-Shaped Sponge-tip Electrode.

J Vis Exp 2019 Mar 27(145). Epub 2019 Mar 27.

Department of Optometry and Vision Sciences, University of Melbourne;

The zebrafish (Danio rerio) is commonly used as a vertebrate model in developmental studies and is particularly suitable for visual neuroscience. For functional measurements of visual performance, electroretinography (ERG) is an ideal non-invasive method, which has been well established in higher vertebrate species. This approach is increasingly being used for examining the visual function in zebrafish, including during the early developmental larval stages. Read More

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http://dx.doi.org/10.3791/59487DOI Listing

Fluorescence Recovery after Photobleaching of Yellow Fluorescent Protein Tagged p62 in Aggresome-like Induced Structures.

J Vis Exp 2019 Mar 26(145). Epub 2019 Mar 26.

Department of Molecular Pharmacology and Therapeutics, Loyola University Chicago;

Fluorescence recovery after photobleaching (FRAP) is a microscopy technique that can be used to quantify protein mobility in live cells. In a typical FRAP experiment, steady-state fluorescence is observed by repeated imaging with low-intensity laser light. Subsequently, the fluorescent molecules are rapidly and irreversibly impaired via brief exposure to high-intensity laser light. Read More

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http://dx.doi.org/10.3791/59288DOI Listing

Confocal Live Imaging of Shoot Apical Meristems from Different Plant Species.

Authors:
Yuan Geng Yun Zhou

J Vis Exp 2019 Mar 29(145). Epub 2019 Mar 29.

Department of Botany and Plant Pathology, Purdue University; Purdue Center for Plant Biology, Purdue University;

The shoot apical meristem (SAM) functions as a conserved stem cell reservoir and it generates almost all aboveground tissues during the postembryonic development. The activity and morphology of SAMs determine important agronomic traits, such as shoot architecture, size and number of reproductive organs, and most importantly, grain yield. Here, we provide a detailed protocol for analyzing both the surface morphology and the internal cellular structure of the living SAMs from different species through laser scanning confocal microscope. Read More

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https://www.jove.com/video/59369/confocal-live-imaging-shoot
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http://dx.doi.org/10.3791/59369DOI Listing
March 2019
1 Read

Setup and Execution Of the Blindfolded Code Training Exercise.

J Vis Exp 2019 Mar 29(145). Epub 2019 Mar 29.

Department of Emergency Medicine, Indiana University School of Medicine;

Miscommunication is the most common cause of preventable patient harm in medicine. Currently, there is limited knowledge of innovative techniques to improve resident physician communication and leadership strategies in high-acuity situations. The blindfolded code training exercise removes visual stimuli from the team leader, forcing the team leader to effectively utilize closed-loop communication. Read More

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https://www.jove.com/video/59248/setup-and-execution-of-the-
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http://dx.doi.org/10.3791/59248DOI Listing
March 2019
1 Read

Measuring Enzymatic Stability by Isothermal Titration Calorimetry.

J Vis Exp 2019 Mar 26(145). Epub 2019 Mar 26.

Department of Nutrition, Dietetics and Food Science, Brigham Young University;

This work demonstrates a new method for measuring the stability of enzyme activity by isothermal titration calorimetry (ITC). The peak heat rate observed after a single injection of the substrate solution into an enzyme solution is correlated with enzyme activity. Multiple injections of the substrate into the same enzyme solution over time show the loss of enzyme activity. Read More

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http://dx.doi.org/10.3791/59302DOI Listing

Intense Pulsed Light for the Treatment of Dry Eye Owing to Meibomian Gland Dysfunction.

J Vis Exp 2019 Apr 1(146). Epub 2019 Apr 1.

Carones Ophthalmology Center.

Dry eye disease (DED) is an increasingly common condition and one of the most common complaints of patients. The vast majority of DED is caused by the so-called "evaporative" subtype, that is mainly caused by meibomian gland dysfunction (MGD). Intense pulsed light (IPL) devices employ high intensity pulses of polychromatic lights with a broad range of wavelength (515-1200 nm). Read More

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http://dx.doi.org/10.3791/57811DOI Listing
April 2019
2 Reads

Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis.

J Vis Exp 2019 Mar 30(145). Epub 2019 Mar 30.

School of Basic Pharmaceutical and Toxicological Sciences, College of Pharmacy, University of Louisiana Monroe;

Lung cancer (LC) is the leading cause of cancer-related deaths worldwide. Similar to other cancer cells, a fundamental characteristic of LC cells is unregulated proliferation and cell division. Inhibition of proliferation by halting cell cycle progression has been shown to be a promising approach for cancer treatment, including LC. Read More

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https://www.jove.com/video/59460/analysis-combinatorial-mirn
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http://dx.doi.org/10.3791/59460DOI Listing
March 2019
2 Reads

A Bedside, Single Burr Hole Approach to Multimodality Monitoring in Severe Brain Injury.

J Vis Exp 2019 Mar 26(145). Epub 2019 Mar 26.

Department of Neurosurgery, University of Cincinnati; Neurotrauma Center, University of Cincinnati Gardner Neuroscience Institute.

Intracranial pressure (ICP) monitoring is a cornerstone of the intensive care management of patients with severe acute brain injuries, including traumatic brain injury. While elevations in ICP are common, data regarding the measurement and treatment of these ICP elevations are conflicting. There is increasing recognition that changes in the balance between supply and demand of brain tissue are critically important and therefore the measurement of multiple modalities is required. Read More

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https://www.jove.com/video/58993/a-bedside-single-burr-hole-
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http://dx.doi.org/10.3791/58993DOI Listing
March 2019
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Efficient Generation of Pancreas/Duodenum Homeobox Protein 1+ Posterior Foregut/Pancreatic Progenitors from hPSCs in Adhesion Cultures.

J Vis Exp 2019 Mar 27(145). Epub 2019 Mar 27.

Center for iPS Cell Research and Application (CiRA), Kyoto University;

Human pluripotent stem cell (hPSC)-derived pancreatic cells are a promising cell source for regenerative medicine and a platform to study human developmental processes. Stepwise directed differentiation that recapitulates developmental processes is one of the major ways to generate pancreatic cells including pancreas/duodenum homeobox protein 1 (PDX1) pancreatic progenitor cells. Conventional protocols initiate the differentiation with small colonies shortly after the passage. Read More

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http://dx.doi.org/10.3791/57641DOI Listing

Visualization of the Superior Ocular Sulcus during Danio rerio Embryogenesis.

J Vis Exp 2019 Mar 27(145). Epub 2019 Mar 27.

Department of Biological Sciences, University of Alberta; Women & Children's Health Research Institute, University of Alberta; Neuroscience and Mental Health Research Institute, University of Alberta;

Congenital ocular coloboma is a genetic disorder that is typically observed as a cleft in the inferior aspect of the eye resulting from incomplete choroid fissure closure. Recently, the identification of individuals with coloboma in the superior aspect of the iris, retina, and lens led to the discovery of a novel structure, referred to as the superior fissure or superior ocular sulcus (SOS), that is transiently present on the dorsal aspect of the optic cup during vertebrate eye development. Although this structure is conserved across mice, chick, fish, and newt, our current understanding of the SOS is limited. Read More

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https://www.jove.com/video/59259/visualization-superior-ocul
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http://dx.doi.org/10.3791/59259DOI Listing
March 2019
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Fully Autonomous Characterization and Data Collection from Crystals of Biological Macromolecules.

J Vis Exp 2019 Mar 22(145). Epub 2019 Mar 22.

Grenoble Outstation, European Molecular Biology Laboratory;

High-brilliance X-ray beams coupled with automation have led to the use of synchrotron-based macromolecular X-ray crystallography (MX) beamlines for even the most challenging projects in structural biology. However, most facilities still require the presence of a scientist on site to perform the experiments. A new generation of automated beamlines dedicated to the fully automatic characterization of, and data collection from, crystals of biological macromolecules has recently been developed. Read More

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http://dx.doi.org/10.3791/59032DOI Listing

A Reliable and Reproducible Critical-Sized Segmental Femoral Defect Model in Rats Stabilized with a Custom External Fixator.

J Vis Exp 2019 Mar 24(145). Epub 2019 Mar 24.

Department of Orthopedics and Rehabilitation, University of Wisconsin-Madison.

Orthopedic research relies heavily on animal models to study mechanisms of bone healing in vivo as well as investigate the new treatment techniques. Critical-sized segmental defects are challenging to treat clinically, and research efforts could benefit from a reliable, ambulatory small animal model of a segmental femoral defect. In this study, we present an optimized surgical protocol for the consistent and reproducible creation of a 5 mm critical diaphyseal defect in a rat femur stabilized with an external fixator. Read More

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http://dx.doi.org/10.3791/59206DOI Listing

Combining Histochemical Staining and Image Analysis to Quantify Starch in the Ovary Primordia of Sweet Cherry during Winter Dormancy.

J Vis Exp 2019 Mar 20(145). Epub 2019 Mar 20.

Instituto Agroalimentario de Aragón - IA2 (CITA-Universidad de Zaragoza), Centro de Investigación y Tecnología Agroalimentaria de Aragón.

Changes in starch in small structures are associated with key events during several plant developmental processes, including the reproductive phase from pollination to fertilization and the onset of fruiting. However, variations in starch during flower differentiation are not completely known, mainly due to the difficulty of quantifying the starch content in the particularly small structures of the flower primordia. Here, we describe a method for the quantification of starch in the ovary primordia of sweet cherry (Prunus avium L. Read More

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http://dx.doi.org/10.3791/58524DOI Listing

Infrared Degenerate Four-wave Mixing with Upconversion Detection for Quantitative Gas Sensing.

J Vis Exp 2019 Mar 22(145). Epub 2019 Mar 22.

Combustion Physics, Department of Physics, Lund University.

We present a protocol for performing gas spectroscopy using infrared degenerated four-wave mixing (IR-DFWM), for the quantitative detection of gas species in the ppm-to-single-percent range. The main purpose of the method is the spatially resolved detection of low-concentration species, which have no transitions in the visible or near-IR spectral range that could be used for detection. IR-DFWM is a nonintrusive method, which is a great advantage in combustion research, as inserting a probe into a flame can change it drastically. Read More

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http://dx.doi.org/10.3791/59040DOI Listing
March 2019
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Dynamic Inter-subject Functional Connectivity Reveals Moment-to-Moment Brain Network Configurations Driven by Continuous or Communication Paradigms.

J Vis Exp 2019 Mar 21(145). Epub 2019 Mar 21.

Institute of Bioengineering, École Polytechnique Fédérale de Lausanne (EPFL); Department of Radiology and Medical Informatics, University of Geneva.

Task-based functional magnetic resonance imaging bears great potential to understand how our brain reacts to various types of stimulation; however, this is often achieved without considering the dynamic facet of functional processing, and analytical outputs typically account for merged influences of task-driven effects and underlying spontaneous fluctuations of brain activity. Here, we introduce a novel methodological pipeline that can go beyond these limitations: the use of a sliding-window analytical scheme permits tracking of functional changes over time, and through cross-subject correlational measurements, the approach can isolate purely stimulus-related effects. Thanks to a rigorous thresholding process, significant changes in inter-subject functional correlation can be extracted and analyzed. Read More

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http://dx.doi.org/10.3791/59083DOI Listing