Drosophila melanogaster (fruit fly)

Due to their large size and relatively simple organization, Drosophila nurse cells are ideally suited for live cell imaging studies. This video describes a microinjection method for exogenous compounds delivery into the cells, and the featured protocol demonstrates the procedure with fluorescent oligonucleotide probes called molecular beacons (MBs) used to visualize endogenous mRNA transcripts.

The Drosophila embryo is surrounded by protective outer membranes. To gain access to the embryo, the chorion–the outermost of the membranes–is often removed. This video describes a commonly used method of dechorionation using sodium hypochlorite. The example protocol demonstrates the procedure while employing sterile techniques necessary for axenic or gnotobiotic animal rearing.

Immunostaining of Drosophila late-stage oocytes can prove challenging due to the oocytes' surrounding membranes. This video describes a high-throughput method for obtaining oocytes, their fixation, and membranes' removal for immunostaining. The featured protocol demonstrates these techniques with late-stage oocytes used to visualize different stages of meiosis.

This video describes a method to mechanically remove the chorion and vitelline membranes from previously fixed mature Drosophila oocytes. The featured protocol shows a detailed demonstration of the procedure yielding oocytes compatible with fluorescent in situ hybridization assays.

This video describes how to perform microinjections that deliver reagents into live Drosophila embryos. Researchers can use this method to introduce exogenous compounds like nucleic acids and proteins during the early stages of embryo development. The featured protocol demonstrates how to set up and perform the procedure for injection of any soluble reagent–here, a fluorescent protein for live imaging experiments.

Source:Siddarth Gunnala1, Nafisa M. Jadavji2,3,41Department of Biomedical Sciences, Midwestern University, Glendale, AZ, USA2Division of Integrative and Molecular Physiology, Department of Biomedical Sciences, Southern University of Illinois, Carbondale, IL, USA3Department of Child Health, College of Medicine Phoenix, University of Arizona, Phoenix, AZ, USA4Department of Neuroscience, Carleton University, Ottawa, ON, CanadaThis video demonstrates hypoxia-induced climbing behavior in Drosophila...

This video shows a method to test hypoxia in Drosophila larvae by assessing their burrowing and tunneling abilities. Using this technique researchers can distinguish larvae experiencing oxygen deprivation from those with developmental deficits. The example protocol demonstrates the setup for this assay.

When staining Drosophila imaginal discs, often a rough dissection is performed where the discs are exposed but remain attached to the body. Once stained, the discs of interest are isolated and mounted. The example protocol demonstrates a procedure for wing imaginal disc dissection and mounting.

The Drosophila neuromuscular junction (NMJ) is used as a model to study the morphology and function of synapses. This video describes important features that researchers quantify to characterize the NMJ. The example protocol demonstrates a software able to perform the quantification automatically.

This video describes two-photon laser ablation of axons in a Drosophila melanogaster larva and an example protocol to induce injury and image the injury site.

This video describes how to prepare larval fillets used to visualize sensory dendritic arborization (da) neurons and their associated epidermal cells. The featured protocol shows in detail how to remove muscle tissue from the body wall while preserving the cells' morphology, which improves immunostaining and visualization of the otherwise obscured da neurons and epidermal cells.

The Drosophila hemolymph serves as both blood and interstitial fluid, circulating throughout the body during the larva and adult stages. This video describes a protocol to collect larval hemolymph in order to quantify the circulating cells.

Drosophila larvae are light-sensitive and respond to changes in surrounding light. This video describes an assay that tests their ability to avoid light, called the light spot assay. The featured protocol demonstrates how to set up the assay with a blue light LED and shows how to record the animal's behavior in response to the light encounter.

This video describes the Drosophila ring gland, a complex endocrine structure important for biosynthesis and secretion of ecdysteroids and other hormones. The featured protocol demonstrates its dissection, fixation, and immunostaining.

Drosophila larvae have dedicated sensory neurons–nociceptors–whose response to potentially harmful thermal stimuli triggers aversive–or so-called "nocifensive"–behaviors in the animal. This video describes a method to assess thermal nociception by recording larval behavior in response to local application of a heat probe onto the cuticle. This method is, therefore, called the local heat probe assay.

Drosophila fruit flies can distinguish small changes in temperature and therefore select environments with favorable thermal conditions. This video describes a behavioral assay that tests the temperature preference of Drosophila larvae on a linear thermal gradient.

This video describes the Drosophila lymph gland—the primary site of hematopoiesis in the fly larva—and shows an example protocol to dissect and mount the organ for immunohistochemistry and fluorescent imaging.

This article describes a technique to remove an insect’s puparium while keeping the inside pupa intact and therefore fit for further investigation. The example protocol shows the procedure with Drosophila guttifera; however, this technique is applicable to other Drosophila species.

This video describes how to dissect and isolate the eye-brain complex from Drosophila pupae. The featured protocol demonstrates the procedure yielding high-quality tissue, compatible with, for example, immunostaining, as well as, other gene expression experiments.

The Drosophila flight muscles are used as a model system for muscle development and physiology. This video describes the flight musculature of adults and highlights a protocol to dissect developing indirect flight muscles that are suitable for RNA sequencing from pupae.

This video describes a classical conditioning assay that tests learning and memory in Drosophila, called courtship conditioning. The assay is based on the reduction of male courtship after experiencing a rejection by a non-receptive premated female. The example protocol shows a setup for the procedure that can be used to assess short- and long-term memory.

This video describes a simple method to quantify fly locomotion and shows the climbing assay performed.  The fly climbing assay is a general test of motor function which relies on a negative geotaxis response.

This video describes the dissection of the accessory glands from male flies and shows an example in which the isolated gland will be dissociated into a single cell suspension.

This video describes the general anatomy of the Drosophila ovary and features a method of dissecting ovaries for live or fixed imaging.

This article describes the dissection of the anterior Malpighian tubules (MT) from a female Drosophila fly and an example protocol from Rossano et al. (2017) in which the MTs are prepared for live imaging in a perfusion chamber.

This video describes the island assay, a behavioral method used to study Drosophila locomotion. The sample protocol features a setup for the assay compatible with semi-automated image analysis.

Scientists use the Drosophila lifespan protocol to measure how different experimental conditions affect flies' longevity and generate survival curves in the process to quantify the experimental outcomes. The sample protocol features the assay being applied to study the effects on longevity from exposure to varying concentrations of endocrine disruptor chemicals, or EDCs, such as 17-α-ethinylestradiol (EE2).

This video describes a feeding assay for fruit flies in which colored food is used to monitor feeding and the effect added compounds may have on feeding activity. The example protocol tests the effects of several endocrine disrupting chemicals (EDC).

This video describes a method of mounting intact brains from adult flies under a bridge slide and shows the protocol performed on immunostained brains for fluorescent microscopy.

This video describes a screening method to identify transgenic Drosophila flies, whose genome was modified using CRISPR-Cas9. CRISPR-Cas9 is a powerful genome-editing tool that revolutionized the way scientists can manipulate an organism's genome. In the example protocol, we will see how to identify CRISPR-generated mutants, in which an insertion of a transactivation sequence replaces the first exon of the branchless (bnl) gene, thus creating a bnl gene-specific driver line, bnl-LexA.

Scanning electron microscopy (SEM) is a powerful tool to image—primarily—the surface features of a biological sample. Proper sample preparation, however, is critical to obtaining quality images. This video focuses on the dehydration and drying steps of preparing intact adult Drosophila for SEM.

This video describes how to embed Drosophila heads in paraffin using the collar method, a technique that prepares fly heads for serial sectioning. Paraffin sections are used for histological examinations of the brain's anatomy and for the assessment of neurodegeneration. The featured protocol demonstrates how multiple flies can be processed in one preparation, thereby increasing time-efficiency and minimizing experimental artifacts.

This video describes how to perform an injection of live adult flies and includes an example protocol in which labeled particles and trypan blue are injected in fly abdomens to assay phagocytosis.

This video describes the taste preference assay, a behavioral method used to measure attraction or avoidance towards colored solutions that taste differently by assessing the fly's abdominal coloration after ingestion of the preferred substance. The featured protocol demonstrates the procedure used to measure flies' preference towards solutions of varying sucrose concentrations.

This video describes the CApillary FEeder, or CAFE, assay, a behavioral method that measures food intake and preference of unrestrained fruit flies. The featured protocol shows how to perform the assay with minimal evaporation from the liquid-filled capillaries used to deliver the food and track its intake.

This video describes how to dissect and isolate the adult Drosophila brain–a procedure necessary for visualizing the organ, which is otherwise obscured by the cuticle, eye, and tracheal tissue. The example protocol shows a detailed demonstration yielding high-quality preparations that can be used for immunostaining and imaging methods in Drosophila neurobiology.

Drosophila microbiota impacts the animal's development, immunity and physiology. This video describes a method to quantify bacteria from flies by determining the colony forming units from whole body preparations. The featured protocol demonstrates the technique with flies, reared under gnotobiotic conditions using four bacterial species.

This video describes a method to dissect, fix, and mount the Drosophila adult leg while preserving its neuromusculature intact for imaging analysis.

This video describes how to track freely walking flies to assess their general locomotion ability. The featured protocol demonstrates the assay's setup and data analysis using video tracking software.

This video describes one method to build a fly aspirator, which can be used to transfer insects between enclosures without anesthesia. The aspirator in the example protocol was prepared for an experiment to quantify individual grooming...

This video describes in vivo calcium imaging in Drosophila neurons using GCaMP. The featured protocol clip shows how to record changes in GCaMP fluorescence from mushroom body neurons during an olfactory conditioning experiment.

Fruit flies have a complex olfactory system, can discriminate hundreds of odorants, and use their sense of smell to direct behavioral decisions. This video describes the four-way olfactometer assay, a method to study olfactory behavioral responses in flies. The featured protocol clip shows how to handle test flies and what are critical steps while performing the assay.

Aggression is an innate behavior and previous fighting experience affects fighting strategy and outcome of later contests in flies. This video describes an assay used to study Drosophila's aggressive behavior. The example protocol features the assay performed in a setup that eliminates fly handling and therefore reduces stress for the animals, which would otherwise impact the subjects' performance.

This video describes a method to test olfactory exploratory behavior in the fruit fly, called the Y-maze. The featured protocol clip demonstrates how to set up and conduct the assay.

This video describes the Drosophila activity monitor (DAM) system used to track locomotor activity. Researchers use activity data collected from the DAM to study circadian rhythms in fruit flies. The featured protocol clip shows how to load flies in the device and record activity data for circadian experiments.

Optogenetics enables the use of light to manipulate neurons that are genetically engineered to express specific opsins–light-sensitive proteins whose light activation triggers a change in the state of the neuron. Here we describe an optogenetic approach in Drosophila using the opsin Channelrhodopsin2. The example protocol features an optogenetics assay set to study the neuronal circuitry behind the fly's escape behavior.

In this video, we highlight a dissection of the adult Drosophila intestine and collection of midguts suitable for single-cell dissociation and FACS in order to isolate intestinal stem cells.

This video describes the fly's adult heart, or otherwise called the Drosophila dorsal vessel. The featured protocol demonstrates a dissection procedure for live heart preparations.

Drosophila females choose between substrates and select sites that are attractive for egg-laying. This video describes a behavioral assay that tests their decision-making, called the egg-laying preference assay. The featured clip demonstrates the procedure with a custom-built setup, which enables researchers not only to test the behavior of many females at the same time but is also compatible with video recording and optogenetic setups.