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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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Drosophila melanogaster: A species of fruit fly much used in genetics because of the large size of its chromosomes.
 JoVE General

Fluorescent Labeling of Drosophila Heart Structures


JoVE 1423 10/13/2009

1,2, 2, 2, 2, 2, 1,2

1Biology Department, San Diego State University, 2Development and Aging Program, NASCR Center, The Sanford Burnham Institute for Medical Research

Here we describe a basic protocol for fluorescent labeling of different elements of heart tubes from larva and adult Drosophila melanogaster. These specimens are well-suited for imaging via fluorescent or confocal microscopy. This technique permits detailed structural analysis of the features of the hearts from a powerful model organism.

 JoVE General

Dissection of Oenocytes from Adult Drosophila melanogaster


JoVE 2242 7/18/2010

,

Department of Biology, University of Toronto

In insects, the oenocytes produce cuticular hydrocarbon compounds. These compounds protect against desiccation and facilitate chemical communication. Here we demonstrate a dissection technique used to isolate the oenocytes from adult Drosophila melanogaster, and illustrate how this preparation can be utilized to study genes involved in hydrocarbon synthesis.

 JoVE General

Paraffin-Embedded and Frozen Sections of Drosophila Adult Muscles


JoVE 2438 12/27/2010

, , , , ,

Gene Expression and Signaling Research Group, Max Planck Institute for Biophysical Chemistry

Identification of mechanisms underlying muscle damage is crucial. Here we present the histological technique for preparing paraffin-embedded and frozen sections of Drosophila thoracic muscles. This allows analysis of muscle morphology and localization of protein and other muscle cell components.

 JoVE Neuroscience

Magnetic Resonance Spectroscopy of live Drosophila melanogaster using Magic Angle Spinning


JoVE 1710 4/15/2010

1,2,3, 2,4, 2,4, 1,2,3

1NMR Surgical Laboratory, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, 2Shriners Burn Institute, 3Department of Radiology, Athinoula A. Martinos Center of Biomedical Imaging, Harvard Medical School, 4Molecular Surgery Laboratory, Department of Surgery, Massachusetts General Hospital, Harvard Medical School

This technique enables the use of high-resolution magic angle spinning proton MR spectroscopy (HRMAS 1H-MRS) for molecular characterization of live Drosophila melanogaster with a conventional 14.1 tesla spectrometer equipped with an HRMAS probe.

 JoVE General

Measurement of Lifespan in Drosophila melanogaster


JoVE 50068 1/07/2013

1, 1, 2, 1

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

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 Neuroscience

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae


JoVE 1725 2/17/2010

, ,

Laboratory of Neurogenetics and Behavior, Rockefeller University

Electrophysiological responses of olfactory sensory neurons to odorants can be measured in insects using single sensillum recordings. In this video article we will demonstrate how to perform single sensillum recordings in the antennae of the vinegar fly (Drosophila melanogaster) and the maxillary palps of the malaria mosquito (Anopheles gambiae).

 JoVE Neuroscience

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster


JoVE 2412 1/14/2011

1, 2, 1

1Institute of Healthy Ageing, and GEE, University College London - UCL, 2School of Biosciences, University of Kent

The Giant Fiber System is a simple neuronal circuit of adult Drosophila melanogaster containing the largest neurons in the fly. We describe the protocol for monitoring synaptic transmission through this pathway by recording post synaptic potentials in dorsal longitudinal (DLM) and tergotrochanteral (TTM) muscles following direct stimulation of the Giant Fiber interneurons.

 JoVE Immunology and Infection

RNAi Screening for Host Factors Involved in Vaccinia Virus Infection using Drosophila Cells


JoVE 2137 8/25/2010

, ,

Department of Microbiology, Penn Genome Frontiers Institute, University of Pennsylvania

Novel host factors involved in viral infection can be identified through cell-based genome-wide loss of function RNAi screening. A Drosophila cell culture model is particularly amenable to this approach due to the ease and efficiency of RNAi. Here we demonstrate this technique using vaccinia virus as an example.

 JoVE General

In vivo Imaging of Intact Drosophila Larvae at Sub-cellular Resolution


JoVE 2249 9/10/2010

*1,2, *1, 1,2, 1, 1, 1

1Junior Research Group Synaptic Plasticity, Hertie Institute for Clinical Brain Research, University of Tübingen, 2Graduate School of Cellular and Molecular Neuroscience, University of Tübingen

This protocol describes a reliable method for anesthetization and imaging of intact Drosophila melanogaster larvae. We have utilized the volatile anesthetic desflurane to allow for repetitive imaging at sub-cellular resolution and re-identification of structures for up to a few days1.

 JoVE Neuroscience

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila


JoVE 2157 9/28/2010

1,2, 1,3, 1, 1,3, 1,3

1Center for Advanced Biotechnology and Medicine, Rutgers University, 2Current Address: Department of Entomology, College of Agricultural and Environmental Sciences, University of California, Davis, 3Department of Molecular Biology and Biochemistry, Rutgers University

We describe procedures for recording daily locomotor activity rhythms of Drosophila and subsequent data analysis. Locomotor activity rhythms are a reliable behavioral output of animal circadian clocks and are used as the standard readout of clock function when studying circadian mutants or examining how the environment regulates the circadian system.

 JoVE Immunology and Infection

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


JoVE 3347 5/07/2012

*1,2, *1,2, 1, 1,2

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

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 Editorial

May 2011: This Month in JoVE


JoVE 3449 5/04/2011

The main highlights for our May issue include methods for measuring cognition in zero gravity, isolating mosquito immune cells, engineering recombinant SARS vaccines, and detecting tumors with thermal imaging. In addition, procedures for isolating neural stem cells from human fetal brain and culturing antigen-presenting liver cells will also be released.

 JoVE Neuroscience

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster


JoVE 50192 1/25/2013

,

Department of Neurobiology, Stanford University

Genetically encoded optogenetic tools enable noninvasive manipulation of specific neurons in the Drosophila brain. Such tools can identify neurons whose activation is sufficient to elicit or suppress particular behaviors. Here we present a method for activating Channelrhodopsin2 that is expressed in targeted neurons in freely walking flies.

 JoVE General

Isolation of Drosophila melanogaster Testes


JoVE 2641 5/13/2011

,

Department of Biochemistry & Molecular Pharmacology and Howard Hughes Medical Institute, University of Massachusetts Medical School

Drosophila melanogaster testes can be rapidly and efficiently isolated from adult males using dissecting needles. With practice, one can readily isolate in one or two days an amount of testes sufficient for the analysis of DNA or RNA by high throughput sequencing or more traditional molecular biology methods or of protein for antibody- or enzyme-based assays.

 JoVE General

A Magnetic Tether System to Investigate Visual and Olfactory Mediated Flight Control in Drosophila


JoVE 1063 11/21/2008

,

Department of Physiological Science, University of California, Los Angeles

Here we describe how to tether a fly in an olfactory magnetic-tether (OMT) apparatus. We describe how to align the rare-earth magnets and odor ports, and how to set mass flow rates for both the stimulus delivery and vacuum suction to achieve optimal odor tracking.

 JoVE Neuroscience

Assessing Neurodegenerative Phenotypes in Drosophila Dopaminergic Neurons by Climbing Assays and Whole Brain Immunostaining


JoVE 50339 4/24/2013

,

Department of Biomolecular Genetics, University of Rochester Medical Center

Here we describe two assays that have been established to study age-dependent neurodegeneration of dopaminergic (DA) neurons in Drosophila: the climbing/startle-induced negative geotaxis assay which allows to study the functional effects of DA neurons degeneration and the tyrosine hydroxylase immunostaining which is used to identify and count DA neurons in whole brain mounts.

 JoVE Immunology and Infection

Quantitative Measurement of the Immune Response and Sleep in Drosophila


JoVE 4355 12/04/2012

, ,

Center for Sleep and Circadian Neurobiology, University of Pennsylvania Perelman School of Medicine

To understand a link between the immune response and behavior, we describe a method to measure locomotor behavior in Drosophila during bacterial infection as well as the ability of flies to mount an immune response by monitoring survival, bacterial load, and real-time activity of a key regulator of innate immunity, NFκB.

 JoVE Neuroscience

Appetitive Associative Olfactory Learning in Drosophila Larvae


JoVE 4334 2/18/2013

1, 1, 1,2, 1, 1,2

1Department of Biology, University of Konstanz, 2Department of Biology, University of Fribourg

Drosophila larvae are able to associate odor stimuli with gustatory reward. Here we describe a simple behavioral paradigm that allows the analysis of appetitive associative olfactory learning.

 JoVE General

Studying Mitotic Checkpoint by Illustrating Dynamic Kinetochore Protein Behavior and Chromosome Motion in Living Drosophila Syncytial Embryos


JoVE 3763 6/14/2012

,

Institute for Cell and Molecular Biosciences, University of Newcastle, United Kingdom

The kinetochore is where the SAC initiates its signal monitoring the mitotic segregation of the sister chromatids. A method is described to visualize the recruitment and turnover of one of the kinetochore proteins and its coordination with the chromosome motion in Drosophila embryos using a Leica laser scanning confocal system.

 JoVE General

Mesoscopic Fluorescence Tomography for In-vivo Imaging of Developing Drosophila


JoVE 1510 8/20/2009

1, 2, 3, 3, 2, 1

1Center for Systems Biology, Massachusetts General Hospital, 2Institute for Biological and Medical Imaging (IBMI), Technical University of Munich and Helmholtz Center Munich, 3Department of Genetics, Harvard Medical School and Howard Hughes Medical Institute

Mesoscopic fluorescence tomography operates beyond the penetration limits of tissue-sectioning fluorescence microscopy. The technique is based on multi-projection illumination and a photon transport description. We demonstrate in-vivo whole-body 3D visualization of the morphogenesis of GFP-expressing wing imaginal discs in Drosophila melanogaster.

 JoVE General

Experimental Manipulation of Body Size to Estimate Morphological Scaling Relationships in Drosophila


JoVE 3162 10/01/2011

1, 2, 2, 1

1Department of Biology & Biochemistry, University of Houston, 2Department of Zoology, Michigan State University

Morphological scaling relationships capture and describe organismal shape. We present a method to measure morphological scaling relationships across the natural range of body sizes in fully metamorphic insects. Using a simple diet manipulation we increase the distribution of trait sizes, permitting the accurate description of how shape and size co-vary.

 JoVE Neuroscience

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


JoVE 3597 4/15/2012

1, 2, 1, 2, 1

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

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

Mapping and Application of Enhancer-trap Flippase Expression in Larval and Adult Drosophila CNS


JoVE 2649 6/03/2011

1, 1, 1, 1, 1,2, 1, 1, 1, 1

1Department of Zoology, University of Oklahoma - Norman, 2Department of Biology, Brandeis University

We describe a Flippase-induced intersectional Gal80/Gal4 repression (FINGR) method, allowing tissue-specific FLP to determine Gal80 expression patterns. Wherever Gal4 and FLP overlap, Gal4 expression is turned on (Gal80 flipped out) or off (Gal80 flipped in). The FINGR method is versatile for clonal analysis and neural circuit mapping.

 JoVE General

Electrophysiological Recording in the Drosophila Embryo


JoVE 1348 5/21/2009

1, 1, 2

1Department of Biological Sciences, University of Illinois, 2Department of Biological Sciences, Vanderbilt University

Electrophysiological recordings from Drosophila embryos allow analyses of developing muscle and neuron electrical properties, as well as characterization of functional synaptogenesis at the glutamatergic neuromuscular junction and central cholinergic and GABAergic synapses.

 JoVE General

Harvesting and Preparing Drosophila Embryos for Electrophysiological Recording and Other Procedures


JoVE 1347 5/20/2009

1, 1, 2

1Department of Biological Sciences, University of Illinois, 2Department of Biological Sciences, Vanderbilt University

This technique exposes the Drosophila embryonic neuromusculature for immunohistochemistry or electrophysiological recording. It is useful for studying early events in neuromuscular development or performing electrophysiology in mutants that cannot hatch.

 JoVE General

Insulin Injection and Hemolymph Extraction to Measure Insulin Sensitivity in Adult Drosophila melanogaster


JoVE 2722 6/30/2011

1, 2

1Department of Biology, State University of New York, 2Allied Health Sciences, University of Connecticut

Conserved insulin signaling pathways found in the fruit fly Drosophila melanogaster make this organism a potential tool for modeling metabolic disorders including type II diabetes. To this end, it is critical to establish physiological assays to effectively measure systemic insulin action in peripheral glucose disposal in the adult fly.

 JoVE General

Chromatin Immunoprecipitation (ChIP) using Drosophila tissue


JoVE 3745 3/23/2012

, ,

Department of Biology, Johns Hopkins University

Recently high-throughput sequencing technology has greatly increased sensitivity of Chromatin Immunoprecipitation (ChIP) experiment and prompted its application using purified cells or dissected tissue. Here we delineate a method to use ChIP technique with Drosophila tissue, which can address the endogenous chromatin state in a well-characterized biological system.

 JoVE Neuroscience

Whole Mount Preparation of the Adult Drosophila Ventral Nerve Cord for Giant Fiber Dye Injection


JoVE 3080 6/04/2011

,

Department of Biological Sciences, Florida Atlantic University

An in vivo dissection of the adult Drosophila ventral nerve cord (VNC) is demonstrated. This particular dissection method causes little damage to the VNC allowing the subsequent labeling of the giant fiber neurons with fluorescent dye for high resolution imaging.

 JoVE General

Isolation and Purification of Kinesin from Drosophila Embryos


JoVE 3501 4/27/2012

, , ,

Department of Developmental and Cell Biology, School of Biosciences, University of California, Irvine

This is a protocol to isolate active full length Kinesin from Drosophila embryos for single-molecule biophysical studies. We show how to collect embryos, make the embryo lysate, and then polymerize microtubules (MTs). Kinesin is purified by immobilizing it on the MTs, spinning down the Kinesin-MT complexes, and then releasing the kinesin from the MTs via ATP addition.

 JoVE General

Electrophysiological Methods for Recording Synaptic Potentials from the NMJ of Drosophila Larvae


JoVE 1109 2/06/2009

,

Department of Physiology and Cellular Biophysics, Columbia University College of Physicians and Surgeons

Here we describe electrophysiological methods for measuring synaptic transmission at the neuromuscular junction of Drosophila larva. Evoked release is initiated artificially by stimulating the motor neuron axons, and transmission through the NMJ can be measured by the postsynaptic response evoked in the muscle.

 JoVE General

Isolation and Purification of Drosophila Peripheral Neurons by Magnetic Bead Sorting


JoVE 1599 12/01/2009

1,2, 1,2, 1,2, 1,2

1Department of Molecular and Microbiology, George Mason University, 2Krasnow Institute for Advanced Study, George Mason University

In this video-article we present a method for the isolation and purification of Drosophila peripheral neurons using a fast magnetic bead assisted cell sorting strategy. RNA obtained from the isolated cells can be readily used for downstream applications including microarray analyses.

 JoVE General

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


JoVE 50239 5/19/2013

*, *, *, ,

Molecular, Cellular and Developmental Biology, University of Michigan

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.

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