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  JoVE Biology

  
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  JoVE Neuroscience

  
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  JoVE Immunology and Infection

  
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  JoVE Clinical and Translational Medicine

  
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  JoVE Bioengineering

  
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  JoVE Applied Physics

  
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  JoVE Chemistry

  
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  JoVE Behavior

  
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  JoVE Environment

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JoVE Science Education

General Laboratory Techniques

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Basic Methods in Cellular and Molecular Biology

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Model Organisms I

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Model Organisms II

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Essentials of
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 JoVE Biology

Alternative Cultures for Human Pluripotent Stem Cell Production, Maintenance, and Genetic Analysis

1NIH Stem Cell Unit, National Institute of Neurological Disorders and Stroke, National Institutes of Health, 2Craniofacial and Skeletal Diseases Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health


JoVE 51519

Here, we present human pluripotent stem cell (hPSC) culture protocols, based on non-colony type monolayer (NCM) growth of dissociated single cells. This new method, utilizing Rho-associated kinase inhibitors or the laminin isoform 521 (LN-521), is suitable for producing large amounts of homogeneous hPSCs, genetic manipulation, and drug discovery.

 JoVE Biology

Rapid Genetic Analysis of Epithelial-Mesenchymal Signaling During Hair Regeneration

1Program in Epithelial Biology, Stanford University School of Medicine


JoVE 4344

Tissue-specific analysis of a hair follicle regeneration model using lentivirus to mediate gain- or loss-of-function.

 JoVE Biology

Mating and Tetrad Separation of Chlamydomonas reinhardtii for Genetic Analysis

1Boyce Thompson Institute for Plant Research, Cornell University


JoVE 1274

Mating and tetrad separation are required for genetic analysis in Chlamydomonas reinhardtii. Here we demonstrate standard methods for gametogenesis, mating, zygote germination and tetrad dissection. This protocol consists of an easy-to-follow series of steps that will make genetic approaches amenable to scientists who are less familiar with Chlamydomonas.

 JoVE Immunology and Infection

Genetic Manipulation in Δku80 Strains for Functional Genomic Analysis of Toxoplasma gondii

1Department of Microbiology and Immunology, The Geisel School of Medicine at Dartmouth


JoVE 50598

Here we report a method for using type I and type II Δku80 strains of Toxoplasma gondii to efficiently generate targeted gene deletions and gene replacements for functional genomic analysis.

 JoVE Biology

Mapping Bacterial Functional Networks and Pathways in Escherichia Coli using Synthetic Genetic Arrays

1Department of Molecular Genetics, University of Toronto, 2Banting and Best Department of Medical Research, Donnelly Centre, University of Toronto, 3Department of Biochemistry, Research and Innovation Centre, University of Regina


JoVE 4056

Systematic, large-scale synthetic genetic (gene-gene or epistasis) interaction screens can be used to explore genetic redundancy and pathway cross-talk. Here, we describe a high-throughput quantitative synthetic genetic array screening technology, termed eSGA that we developed for elucidating epistatic relationships and exploring genetic interaction networks in Escherichia coli.

 JoVE Environment

Methods for Performing Crosses in Setaria viridis, a New Model System for the Grasses

1Donald Danforth Plant Science Center, 2Boyce Thompson Institute


JoVE 50527

We have developed a methodology for performing crosses in Setaria viridis (S. viridis). The method involves pruning the panicle prior to a hot water treatment to kill viable pollen. Crosses are performed following a well-controlled growth regime and typically result in the recovery of 1 to 7 cross-pollinated seed/s per panicle.

 JoVE Biology

Production of Haploid Zebrafish Embryos by In Vitro Fertilization

1Department of Biological Sciences, University of Notre Dame


JoVE 51708

The zebrafish is a powerful model system for developmental biology and human disease research due to their genetic similarity with higher vertebrates. This protocol describes a methodology to create haploid zebrafish embryos that can be utilized for forward screen strategies to identify recessive mutations in genes essential for early embryogenesis.

 JoVE Clinical and Translational Medicine

A Protocol for Genetic Induction and Visualization of Benign and Invasive Tumors in Cephalic Complexes of Drosophila melanogaster

1Department of Biology, Western Kentucky University


JoVE 50624

Cooperation between an activated oncogene like RASV12 and mutations in cell polarity genes like scribbled, result in tumor growth in Drosophila where tumor cells also display invasive behaviors. Here a simple protocol for the induction and observation of the benign and invasive tumors is presented.

 JoVE Biology

A Quantitative Fitness Analysis Workflow

1Institute for Cell and Molecular Biosciences, Newcastle University Medical School


JoVE 4018

Quantitative Fitness Analysis (QFA) is a complementary series of experimental and computational methods for estimating microbial culture fitnesses. QFA estimates the effect of genetic mutations, drugs or other applied treatments on microbe growth. Experiments scaling from focussed analysis of single cultures to thousands of parallel cultures can be designed.

 JoVE Neuroscience

Assessment of Morphine-induced Hyperalgesia and Analgesic Tolerance in Mice Using Thermal and Mechanical Nociceptive Modalities

1Biotechnology and Cellular Signalling, UMR 7242 CNRS, Université de Strasbourg


JoVE 51264

We describe a protocol to examine the development of opioid-induced hyperalgesia and tolerance in mice. Based on the measurement of thermal and mechanical nociceptive responses of naïve and morphine-treated animals, it allows to quantify the increase in pain sensitivity (hyperalgesia) and decrease in analgesia (tolerance) associated with chronic opiate administration.

 JoVE Immunology and Infection

Forward Genetic Approaches in Chlamydia trachomatis

1Department of Molecular Genetics and Microbiology, Center for Microbial Pathogenesis, Duke University Medical Center


JoVE 50636

We describe a methodology to perform genetic analysis in Chlamydia based on chemical mutagenesis and whole genome sequencing. In addition, a system for DNA exchange within infected cells is described that can be used for genetic mapping. This method may be broadly applicable to microbial systems lacking transformation systems and molecular genetic tools.

 JoVE Biology

Measurement of Lifespan in Drosophila melanogaster

1Department of Molecular and Integrative Physiology, University of Michigan, 2Cellular and Molecular Biology Program, University of Michigan


JoVE 50068

Drosophila melanogaster is a powerful model organism for exploring the molecular basis of longevity regulation. This protocol will discuss the steps involved in generating a reproducible, population-based measurement of longevity as well as potential pitfalls and how to avoid them.

 JoVE Biology

Identifying Protein-protein Interaction in Drosophila Adult Heads by Tandem Affinity Purification (TAP)

1Neuroscience Center of Excellence, Louisiana State University Health Sciences Center


JoVE 50968

Drosophila is famous for its powerful genetic manipulation, but not for its suitability of in-depth biochemical analysis. Here we present a TAP-based procedure to identify interacting partners of any protein of interest from the fly brain. This procedure can potentially lead to new avenues of research.

 JoVE Clinical and Translational Medicine

A Neuroscientific Approach to the Examination of Concussions in Student-Athletes

1Department of Exercise Science, Elon University, 2Department of Physical Therapy Education, Elon University, 3Department of Physical Therapy, Duquesne University, 4Department of Sports Medicine, Elon University


JoVE 52046

There is great variability in an individual’s risk for concussion and their corresponding recovery. A multifaceted approach to concussion evaluation is warranted; including baseline testing of athletes before participation in sport and timely evaluation post injury. The goal of this protocol is to provide an appropriate multifaceted approach to examine concussions.

 JoVE Biology

Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay

1Department of Biological and Allied Health Sciences, Fairleigh Dickinson University


JoVE 51203

An egg-in-worm (EIW) assay is a useful method to quantify egg-laying behavior. Alterations in egg laying can be a behavioral response of the model organism Caenorhabditis elegans to potentially harmful environmental substances such as those produced by pathogenic bacteria.

 JoVE Immunology and Infection

A Genetic Screen to Isolate Toxoplasma gondii Host-cell Egress Mutants

1Department of Biology, Boston College


JoVE 3807

Forward genetics is a powerful method to unravel the molecular level of how Toxoplasma egresses from its host cell. Protocols are provided to chemically mutagenize parasites, enrich for mutants with defects in induced egress, and validate the phenotype of cloned mutants.

 JoVE Biology

Examination of Drosophila Larval Tracheal Terminal Cells by Light Microscopy

1Department of Human Genetics, University of Utah


JoVE 50496

Here, we present a method for light microscopy analysis of tracheal terminal cells in Drosophila larvae. This method allows for quick examination of branch and lumen morphology in whole animals and would be useful for analysis of individual mutants or screens for mutations affecting terminal cell development.

 JoVE Biology

RNA Secondary Structure Prediction Using High-throughput SHAPE

1RT Biochemistry Section, HIV Drug Resistance Program, Frederick National Laboratory for Cancer Research


JoVE 50243

High-throughput selective 2' hydroxyl acylation analyzed by primer extension (SHAPE) utilizes a novel chemical probing technology, reverse transcription, capillary electrophoresis and secondary structure prediction software to determine the structures of RNAs from several hundred to several thousand nucleotides at single nucleotide resolution.

 JoVE Biology

Non-enzymatic, Serum-free Tissue Culture of Pre-invasive Breast Lesions for Spontaneous Generation of Mammospheres

1Center for Applied Proteomics and Molecular Medicine, George Mason University, 2Virginia Surgery Associates


JoVE 51926

Primary cell culture using intact tissue organoids provides a model system that mimics the multi-cellular in vivo microenvironment. We developed a serum-free primary breast epithelium tissue culture model that perpetuates mixed cell culture lineages and exhibits differentiated morphology, without enzymatic tissue disruption. Breast organoids remain viable for >6 months.

 JoVE Biology

Dietary Supplementation of Polyunsaturated Fatty Acids in Caenorhabditis elegans

1School of Molecular Biosciences, Washington State University, 2Center for Reproductive Biology, Washington State University


JoVE 50879

Caenorhabditis elegans is a useful model to explore the functions of polyunsaturated fatty acids in development and physiology. This protocol describes an efficient method of supplementing the C. elegans diet with polyunsaturated fatty acids.

 JoVE Biology

Vampiric Isolation of Extracellular Fluid from Caenorhabditis elegans

1Department of Molecular and Cellular Biology, Harvard University


JoVE 3647

The model organism C. elegans uses pseudocoelomic fluid as a passive circulatory system. Direct assay of this fluid has not been previously possible. Here we present a novel technique to directly assay the extracellular space, and use systemic silencing signals during an RNAi response as a proof of principle example.

 JoVE Neuroscience

Labeling of Single Cells in the Central Nervous System of Drosophila melanogaster

1Institute of Genetics, University of Mainz, 2Department of Anatomy and Neuroscience, University of Melbourne


JoVE 50150

We present a technique for labeling single neurons in the central nervous system (CNS) of Drosophila embryos, which allows the analysis of neuronal morphology by either transmitted light or confocal microscopy.

 JoVE Clinical and Translational Medicine

Pharmacologic Induction of Epidermal Melanin and Protection Against Sunburn in a Humanized Mouse Model

1The Markey Cancer Center, University of Kentucky College of Medicine, 2Graduate Center for Toxicology, University of Kentucky College of Medicine, 3Department of Molecular and Biomedical Pharmacology, University of Kentucky College of Medicine, 4Department of Pediatrics, University of Kentucky College of Medicine


JoVE 50670

Epidermal melanin is induced by topical application of forskolin in a murine model of the fair-skinned UV-sensitive human. Pharmacologic manipulation of cAMP levels in the skin and epidermal darkening strongly protect against UV-mediated inflammation (sunburn) as measured by the minimum erythematous dose (MED) assay.

 JoVE Biology

Microgavage of Zebrafish Larvae

1Department of Cell and Molecular Physiology, University of North Carolina at Chapel Hill


JoVE 4434

We present a novel method for microgavage of larval zebrafish utilizing standard embryo microinjection and stereomicroscopy equipment. We demonstrate that microgavage is a safe and efficient technique useful for delivering controlled amounts of diverse materials specifically into the larval zebrafish intestinal lumen.

 JoVE Biology

Cytological Analysis of Spermatogenesis: Live and Fixed Preparations of Drosophila Testes

1Department of Cell and Developmental Biology, Vanderbilt University Medical Center


JoVE 51058

Methods for isolating and preparing Drosophila testes samples (live and fixed) for imaging by phase-contrast and fluorescence microscopy are described herein. 

 JoVE Neuroscience

In vivo Neuronal Calcium Imaging in C. elegans

1Department of Physiology and Biophysics, Boston University School of Medicine, 2Boston University Photonics Center


JoVE 50357

With its small transparent body, well-documented neuroanatomy and a host of amenable genetic techniques and reagents, C. elegans makes an ideal model organism for in vivo neuronal imaging using relatively simple, low-cost techniques. Here we describe single neuron imaging within intact adult animals using genetically encoded fluorescent calcium indicators.

 JoVE Immunology and Infection

Identifying DNA Mutations in Purified Hematopoietic Stem/Progenitor Cells

1Greehey Children's Cancer Research Institute, UT Health Science Center at San Antonio, 2Department of Cellular and Structural Biology, UT Health Science Center at San Antonio, 3Department of Pathology, UT Health Science Center at San Antonio, 4Department of Microbiology, UT Health Science Center at San Antonio, 5Cancer Therapy and Research Center, UT Health Science Center at San Antonio


JoVE 50752

Here we describe an in vivo mutagenesis assay for small numbers of purified hematopoietic cells using the LacI transgenic mouse model. The LacI gene can be isolated to determine the frequency, location, and type of DNA mutants spontaneously arisen or after exposure to genotoxins.

 JoVE Immunology and Infection

Use of Galleria mellonella as a Model Organism to Study Legionella pneumophila Infection

1Center for Molecular Bacteriology and Infection, Imperial College London


JoVE 50964

The larva of the wax moth Galleria mellonella was recently established as an in vivo model to study Legionella pneumophila infection. Here, we demonstrate fundamental techniques to characterize the pathogenesis of Legionella in the larvae, including inoculation, measurement of bacterial virulence and replication as well as extraction and analysis of infected hemocytes.

 JoVE Biology

Imaging Centrosomes in Fly Testes

1Department of Biological Sciences, University of Toledo


JoVE 50938

Imaging of centrosomal proteins during Drosophila spermatogenesis is a powerful method to identify new proteins critical for centrosome biology as well as to elucidate the particular function of known players in this process.

 JoVE Biology

High-throughput Yeast Plasmid Overexpression Screen

1Neuroscience Graduate Group, University of Pennsylvania School of Medicine, 2Department of Cell and Developmental Biology, University of Pennsylvania School of Medicine


JoVE 2836

Here we describe a plasmid overexpression screen in Saccharomyces cerevisiae, using an arrayed plasmid library and a high-throughput yeast transformation protocol with a liquid handling robot.

 JoVE Biology

Reverse Yeast Two-hybrid System to Identify Mammalian Nuclear Receptor Residues that Interact with Ligands and/or Antagonists

1Department of Genetics, Albert Einstein College of Medicine, 2Shanghai Key Laboratory of Complex Prescription and MOE Key Laboratory for Standardization of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine


JoVE 51085

Ketoconazole binds to and antagonizes Pregnane X Receptor (PXR) activation. Yeast high throughput screens of PXR mutants define a unique region for ketoconazole binding. This yeast-based genetic method discovers novel nuclear receptor interactions with ligands that associate with surface binding sites.

 JoVE Biology

The Green Monster Process for the Generation of Yeast Strains Carrying Multiple Gene Deletions

1Department of Synthetic Biology and Bioenergy, J. Craig Venter Institute, 2Department of Microbial and Environmental Genomics, J. Craig Venter Institute, 3Donnelly Centre & Department of Molecular Genetics, University of Toronto, 4Lunenfeld Research Institute, Mt Sinai Hospital


JoVE 4072

The Green Monster method enables the rapid assembly of multiple deletions marked with a reporter gene encoding green fluorescent protein. This method is based on driving yeast strains through repeated cycles of sexual assortment of deletions and fluorescence-based enrichment of cells carrying more deletions.

 JoVE Biology

A Noninvasive Hair Sampling Technique to Obtain High Quality DNA from Elusive Small Mammals

1Department of Biology and Centre for Species at Risk and Habitat Studies,, University of British Columbia, Okanagan Campus


JoVE 2791

We present a noninvasive sampling approach to efficiently collect hair samples from elusive small mammals, as shown for the American pika. We demonstrate the utility of this method by extracting DNA from sampled hair and amplifying several types of molecular markers commonly used in studies of wildlife ecology and conservation.

 JoVE Neuroscience

Paired Nanoinjection and Electrophysiology Assay to Screen for Bioactivity of Compounds using the Drosophila melanogaster Giant Fiber System

1Department of Biological Sciences, Florida Atlantic University, 2Department of Chemistry & Biochemistry, Florida Atlantic University


JoVE 3597

A rapid in vivo assay to test for neuromodulatory compounds using the Giant Fiber System (GFS) of Drosophila melanogaster is described. Nanoinjections in the head of the animal along with electrophysiological recordings of the GFS can reveal bioactivity of compounds on neurons or muscles.

 JoVE Neuroscience

Dissection and Immunohistochemistry of Larval, Pupal and Adult Drosophila Retinas

1Department of Biology, New York University


JoVE 4347

The Drosophila retina is a crystal-like lattice composed of a small number of cell types that are generated in a stereotyped manner 1. Its amenability to sophisticated genetic analysis allows the study of complex developmental programs. This protocol describes dissections and immunohistochemistry of retinas at three discrete developmental stages, with a focus on photoreceptor differentiation.

 JoVE Biology

BEST: Barcode Enabled Sequencing of Tetrads

1Pacific Northwest Diabetes Research Institute


JoVE 51401

Barcode Enabled Sequencing of Tetrads (BEST) replaces the manual processes of isolating, disrupting and spacing tetrads. BEST isolates tetrads by fluorescence-activated cell sorting onto agar plates, separates the spores by agitation with glass beads, and determines which randomly arrayed colonies were derived from the same original tetrad using molecular barcodes.

 JoVE Biology

DNA Extraction from Paraffin Embedded Material for Genetic and Epigenetic Analyses

1Department of Integrative Oncology, BC Cancer Research Centre, 2Interdisciplinary Oncology Program, University of British Columbia - UBC, 3Photography/Video Production, Multi-Media Services, BC Cancer Agency, 4Department of Pathology and Laboratory Medicine, University of British Columbia - UBC


JoVE 2763

This video demonstrates the protocol for DNA extraction from formalin-fixed paraffin-embedded material. This is a multi-day procedure in which tissue sections are deparaffinized with xylene, rehydrated with ethanol and treated with proteinase K to purify and isolate DNA for subsequent gene-specific or genome-wide analysis.

 JoVE Biology

Design and Analysis of Temperature Preference Behavior and its Circadian Rhythm in Drosophila

1The Visual Systems Group, Division of Pediatric Ophthalmology, Cincinnati Childrens Hospital Medical Center, 2PRESTO, JST


JoVE 51097

We recently identified a novel Drosophila circadian output, temperature preference rhythm (TPR), in which the preferred temperature in flies rises during the day and falls during the night. TPR is regulated independently from another circadian output, locomotor activity. Here we describe the design and analysis of TPR in Drosophila.

 JoVE Behavior

Contextual and Cued Fear Conditioning Test Using a Video Analyzing System in Mice

1Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, 2Japan Science and Technology Agency, Core Research for Evolutionary Science and Technology (CREST), 3Center for Genetic Analysis of Behavior, National Institute for Physiological Sciences, National Institutes of Natural Sciences


JoVE 50871

This article presents a protocol for a contextual and cued fear conditioning test using a video analyzing system to assess fear learning and memory in mice.

 JoVE Neuroscience

Light Preference Assay to Study Innate and Circadian Regulated Photobehavior in Drosophila Larvae

1Department of Biology, Institute of Cell and Developmental Biology, University of Fribourg


JoVE 50237

Here we describe a light-dark preference test for Drosophila larva. This assay provides information about innate and circadian regulation of light sensing and processing photobehavior.

 JoVE Neuroscience

In Vivo Imaging of Dauer-specific Neuronal Remodeling in C. elegans

1Department of Crop Sciences, University of Illinois Urbana-Champaign


JoVE 51834

Following exposure to specific environmental stressors, the nematode Caenorhabditis elegans undergoes extensive phenotypic plasticity to enter into a stress-resistant ‘dauer’ juvenile stage. We present methods for the controlled induction and imaging of neuroplasticity during dauer.

 JoVE Immunology and Infection

An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response

1Biology Department, The City College of New York, CUNY, 2The Graduate Center, The City University of New York


JoVE 3347

Parasitoid (parasitic) wasps constitute a major class of natural enemies of many insects including Drosophila melanogaster. We will introduce the techniques to propagate these parasites in Drosophila spp. and demonstrate how to analyze their effects on immune tissues of Drosophila larvae.

 JoVE Immunology and Infection

Automated Separation of C. elegans Variably Colonized by a Bacterial Pathogen

1Department of Integrative Biology, University of California, Berkeley


JoVE 51090

The wormsorter facilitates genetic screens in Caenorhabditis elegans by sorting worms according to expression of fluorescent reporters. Here, we describe a new usage: sorting according to colonization by a GFP-expressing pathogen, and we employ it to examine the poorly understood role of pathogen recognition in initiating immune responses.

 JoVE Biology

Isolating Potentiated Hsp104 Variants Using Yeast Proteinopathy Models

1Department of Biochemistry and Biophysics, Perelman School of Medicine at the University of Pennsylvania


JoVE 52089

Yeast proteinopathy models are valuable tools to assess the toxicity and aggregation of proteins implicated in disease. Here, we present methods for screening Hsp104 variant libraries for toxicity suppressors. This protocol could be adapted to screen any protein library for toxicity suppressors of any protein that is toxic in yeast.

 JoVE Biology

Investigating Protein-protein Interactions in Live Cells Using Bioluminescence Resonance Energy Transfer

1Language and Genetics Department, Max Planck Institute for Psycholinguistics, 2Donders Institute for Brain, Cognition and Behaviour


JoVE 51438

Interactions between proteins are fundamental to all cellular processes. Using Bioluminescence Resonance Energy Transfer, the interaction between a pair of proteins can be monitored in live cells and in real time. Furthermore, the effects of potentially pathogenic mutations can be assessed.

 JoVE Biology

Competitive Genomic Screens of Barcoded Yeast Libraries

1Banting and Best Department of Medical Research and Department of Molecular Genetics, University of Toronto, 2Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 3Donnelly Sequencing Centre, University of Toronto, 4Genetics and Molecular Biology Branch, National Human Genome Research Institute, NIH, 5Stanford Genome Technology Center, Stanford School of Medicine, Stanford University, 6Department of Pharmaceutical Sciences, University of Toronto


JoVE 2864

We have developed comprehensive, unbiased genome-wide screens to understand gene-drug and gene-environment interactions. Methods for screening these mutant collections are presented.

 JoVE Biology

Quantitative and Automated High-throughput Genome-wide RNAi Screens in C. elegans

1Centre d’Immunologie de Marseille-Luminy, Université de la Méditerranée


JoVE 3448

We describe a protocol using C. elegans and RNAi feeding libraries that allows automated measurement of multiple parameters such as fluorescence, size and opacity of individual worms in a population. We give one example of a screen to identify genes involved in anti-fungal innate immunity in C. elegans.

 JoVE Biology

Measuring Caenorhabditis elegans Life Span on Solid Media

1Department of Pathology, University of Washington, 2Molecular and Cellular Biology Program, University of Washington


JoVE 1152

In this article we present a general protocol for measuring life span of nematodes maintained on solid media with UV-killed bacterial food.

 JoVE Biology

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

1Department of Biology, University of Kentucky, Lexington


JoVE 1596

We present various ways to monitor heart function in the larva of Drosophila for assessing questions dealing with the function of gap junctions, ion channel mutations, modulation of pacemaker activity and pharmacological studies.

 JoVE Immunology and Infection

2D and 3D Chromosome Painting in Malaria Mosquitoes

1Department of Entomology, Virginia Tech


JoVE 51173

Chromosome painting is a useful method for studying organization of the cell nucleus and evolution of the karyotype. Here, we demonstrate an approach to isolate and amplify specific regions of interest from single polytene chromosomes that are subsequently used for two- and three-dimensional fluorescent in situ hybridization (FISH).

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