SG est soutenu par des fonds de l&#39;Université d&#39;État de New York à Buffalo et de la Fondation John R. Oishei.

<html> 1. Préparation des réactifs <ol><li> Préparer le Tampon PEMFA l&#39;aide de canaux 100 mM, EGTA 2 mM, et 1 mM MgSO 4. Ajuster le pH du tampon à 7,0. Le tampon est PEMFA filtre stérilisé et stocké à 4 ° C. </li><li> Préparer PBT en ajoutant 1 ml de Triton X-100 à 100ml 1x PBS (Phosphate Buffered Saline) (pH 7,0). Conserver à 4 ° C. </li><li> Préparer 5% de BSA (sérum albumine bovine) avec NaAcide 0,01% dans du PBS 1X (pH 7,0). </li><li> Préparer la solution saline à l&#39;aide de NaCl 0,7% et 0,03% de Triton X-100 dans de l&#39;eau déminéralisée. </li><li> Préparer le fixatif en combinant 4 pièces tampons PEMFA avec 1 partie de formaldéhyde 37%. </li></ol> 2. Collecte d&#39;embryons <ol><li> Préparer une cage pour la souche de drosophile d&#39;intérêt en plaçant&gt; 100 mouches dans une bouteille en plastique contenant un adaptateur d&#39;alimentation avec plaque de gélose. </li><li> Laisser la cage de mouches dans l&#39;obscurité à température ambiante pendant 72 heures, en changeant les plaques toutes les 8-12 heures, de sorte que les mouches s&#39;acclimater à la cage et ne détiennent pas leurs oeufs. </li><li> Pour l&#39;expérience de collecter des embryons pendant 12 heures. Retirer la boîte de Pétri à partir du conteneur et de le remplacer. Prenez la boîte de Pétri enlevé et laver les embryons hors du plat en utilisant la solution saline et un pinceau propre. </li><li> Recueillir les embryons dans une passoire fine. Rincer avec une solution saline pour enlever tout excès de levures. </li><li> En utilisant une spatule délicatement ramasser les embryons et les placer dans un petit flacon de verre avec un capuchon. Ajouter environ 1 ml de solution saline. </li></ol> 3. Retrait des membranes chorion et vitelline et de fixation <ol><li> Après la collecte des embryons, préparer la solution Heptane-Fix en combinant 50% d&#39;heptane et fixateur de 50%. La solution sera en couches avec l&#39;heptane sur le dessus et le fixatif sur le fond. </li><li> Dans le flacon en verre laver les embryons en les incubant dans la solution saline pendant 5 minutes. Retirer l&#39;excès salins aide d&#39;une pipette Pasteur. </li><li> Rincez les embryons avec de l&#39;eau déminéralisée. </li><li> Dechorionate les embryons en les incubant dans l&#39;eau de Javel à 50% pendant 3 minutes. </li><li> Rincez abondamment avec de l&#39;embryon de l&#39;eau déminéralisée. </li><li> Fixer les embryons en les incubant pendant 30 minutes dans la solution d&#39;heptane-Fix en agitant doucement. </li><li> En utilisant une pipette Pasteur remplacer le fixateur (couche inférieure) avec du méthanol en laissant l&#39;heptane dans le tube. Agiter vigoureusement pour enlever la membrane vitelline. Les embryons seront devitellinized couler au fond du tube. </li><li> Recueillir les embryons dans un flacon en verre de nouvelles en utilisant une pipette Pasteur. Rincez les embryons cinq fois avec du méthanol, à chaque fois incuber les embryons pendant 5 minutes. </li></ol> 4. Coloration des anticorps, de montage et de visualisation <ol><li> Réhydrater les embryons dans un mélange de solution PBT 50% et 50% de méthanol pendant 5 minutes. Puis réhydrater en solution PBT 100% pendant 5 minutes. </li><li> Bloquer les embryons en les incubant dans du BSA 5% avec une solution NaAcide 0,01% pendant 1 heure à 4 ° C. </li><li> Les embryons sont maintenant prêts pour la coloration des anticorps. Préparer l&#39;anticorps primaire en le diluant à la concentration appropriée en utilisant la BSA 5% avec une solution NaAcide 0,01%. Nous utilisons un anticorps contre la protéine chaîne de cystéine (CSP), une protéine vésiculaire synaptique et un anticorps anti-HRP ou la peroxydase de raifort, qui reconnaît un neuronaux spécifiques en glucides présents dans le fragment de glycoprotéines de surface et les étiquettes tous les neurones. </li><li> Incuber les embryons nuit à 4 ° C dans la solution d&#39;anticorps primaire. </li><li> Le lendemain, lavez les embryons 10 fois avec PBT au cours d&#39;une heure. </li><li> Préparer la solution d&#39;anticorps secondaire par dilution de la concentration appropriée. Nous utilisons des anticorps Alexa secondaires à une concentration de 1:200. </li><li> Incuber les embryons pendant deux heures dans 4 ° C dans la solution d&#39;anticorps secondaire. </li><li> Laver les embryons 10 fois avec PBT au cours d&#39;une heure. </li><li> Incuber les embryons nuit dans un milieu Vectashield montage. </li><li> Montez les embryons sur une lame de verre et de sceller la lamelle avec du vernis à ongles. Visualisez embryons en utilisant un microscope à fluorescence. </li></ol> 5. Les résultats représentatifs <img src="/files/ftp_upload/2150/2150fig1a.jpg" alt="Figure 1" /> CSP <img src="/files/ftp_upload/2150/2150fig1b.jpg" alt="Figure 1" /> HRP-FITC <img src="/files/ftp_upload/2150/2150fig1c.jpg" alt="Figure 1" /> Fusionnées Figure 1: Un représentant des images montrant le SNC embryonnaire au stade 16-17, utilisant des anticorps contre CSP (rouge) et HRP (vert). Notez la coloration distincte de la commissure. <img src="/files/ftp_upload/2150/2150fig2a.jpg" alt="Figure 2" /> CSP <img src="/files/ftp_upload/2150/2150fig2b.jpg" alt="Figure 2" /> HRP-FITC &quot;/&gt; Fusionnées Figure 2: Un des images représentatives montrant le PNS embryon au stade 16 à 17 en utilisant des anticorps contre CSP (rouge) et HRP (vert). Notez les régularités répétées des neurones périphériques.

<ol>
	<li>Bodmer, R., Barbel, S., Sheperd, S., Jack, J. W., Jan, L. Y., Jan, Y. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transformation+of+sensory+organs+by+mutations+of+the+cut+locus+of+D.+melanogaster.">Transformation of sensory organs by mutations of the cut locus of D. melanogaster.</a> Cell. 51, 293-307 (1987).</li><li>Bodmer, R., Carretto, R., Jan, Y. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurogenesis+of+the+peripheral+nervous+system+in+Drosophila+embryos:+DNA+replication+patterns+and+cell+lineages.">Neurogenesis of the peripheral nervous system in Drosophila embryos: DNA replication patterns and cell lineages.</a> Neuron. 3, 21-32 (1989).</li><li>Campos-Ortega, J. A., Hartenstein, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Embryonic Development of Drosophila melanogaster. , (1985).</li><li>Carroll, S. B., Scott, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Localization+of+the+fushi+tarazu+protein+during+Drosophila+embryogenesis.">Localization of the fushi tarazu protein during Drosophila embryogenesis.</a> Cell. 43, 47-57 (1985).</li><li>Dambly-Chaudi&#232;re, C., Ghysen, A., Jan, Y. N., Jan, L. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+connections+between+imaginal+discs+in+Drosophila.">Muscle connections between imaginal discs in Drosophila.</a> Developmental Biology. 113, 288-294 (1986).</li></ol>

L&#39;embryon de drosophile est un système puissant pour étudier les modifications cellulaires et moléculaires au cours du développement neuronal. Dans ce protocole, nous avons démontré comment préparer embryons monter ensemble pour l&#39;immunohistochimie. Nous avons adapté ce protocole à partir du protocole décrit dans Carroll et Scott, 1985. Ce protocole peut également être adapté pour tester d&#39;autres anticorps neuronale, mais l&#39;optimisation des anticorps devrait être fait. De plus, des...

<table><tbody><tr><td>37% formaldehyde</td><td>Fisher Scientific</td><td>BP531-25</td><td></td></tr><tr><td>CSP</td><td>Developmental Studies Hybridoma Bank</td><td></td><td></td></tr><tr><td>HRP-FITC</td><td>Jackson ImmunoResearch</td><td>323-095-021</td><td></td></tr><tr><td>Alexa Fluor 568 Goat Anti-Mouse IgG</td><td>Invitrogen</td><td>A-11004</td><td></td></tr><tr><td>Vectashield Mounting Medium</td><td>Vector Laboratories</td><td>H-1000</td><td></td></tr></tbody></table>

visualization of the embryonic nervous system in whole-mount drosophila embryos

L&#39;embryon de drosophile est un système modèle intéressant pour étudier les bases cellulaires et moléculaires du développement neuronal. Nous décrivons ici la procédure pour la visualisation de la drosophile embryonnaire du système nerveux en utilisant des anticorps dirigés contre les protéines neuronales. Depuis la toute embryonnaire nerveux périphérique et le système nerveux central sont bien caractérisés au niveau des cellules individuelles (Dambly-Chaudière et al, 1986;.. Bodmer et al, 1987;. Bodmer et al, 1989), les aberrations de ces systèmes peuvent être facilement identifiés à l&#39;aide des anticorps contre différentes protéines neuronales. Les embryons en développement sont collectées à certains moments afin de s&#39;assurer que les embryons sont au stade de développement approprié pour la visualisation. Après la collecte, les couches externes de l&#39;embryon, la membrane chorion et l&#39;enveloppe vitelline qui entoure l&#39;embryon, sont enlevés avant la fixation. Les embryons sont ensuite incubés avec des anticorps neuronale et visualisés en utilisant des anticorps marqués par fluorescence secondaire. Les embryons à des stades de 12 à 17 sont visualisés pour accéder au système embryonnaire nerveux. Au stade 12 la bande de germe de SNC commence le shortening et par l&#39;étape 15 du modèle définitif de la commissure a été atteint. Au stade 17 les contrats SNC et le SNP est entièrement développé (Campos-Ortega et al. 1985). Ainsi, les variations dans la structure de la SNP et du SNC peuvent être facilement observés au cours de ces stades de développement.

Watch this Scientific Journal Video about Visualization of the Embryonic Nervous System in Whole-mount Drosophila Embryos at JoVE.com

Visualization of the Embryonic Nervous System in Whole-mount Drosophila Embryos

Nous décrivons la procédure pour préparer scène<em&gt; Drosophile</em&gt; Embryons pour la visualisation du système nerveux embryonnaire, lors de l&#39;embryogenèse.

La visualisation du système nerveux embryonnaire, en totalité montage Drosophile Embryons

The Drosophila embryo is an attractive model system for investigating the cellular and molecular basis of neuronal development. Here we describe the ...

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Neuroscience

Visualization of the Embryonic Nervous System in Whole-mount Drosophila Embryos

13.2K vues.  SUNY-University at Buffalo. Nous décrivons la procédure pour préparer scène Drosophile Embryons pour la visualisation du système nerveux embryonnaire, lors de l'embryogenèse.

Vidéo: Visualization of the Embryonic Nervous System in Whole-mount Drosophila Embryos

Dissection of Larval CNS in Drosophila Melanogaster

The central nervous system (CNS) of Drosophila larvae is complex and poorly understood.  One way to investigate the CNS is to use immunohistochemistry to examine the expression of various novel and marker proteins.  Staining of whole larvae is impractical because the tough cuticle prevents antibodies from penetrating inside the body cavity.  In order to stain these tissues it is necessary to dissect the animal prior to fixing and staining.  In this article we demonstrate how to dissect Drosophila larvae without damaging the CNS.  Begin by tearing the larva in half with a pair of fine forceps, and then turn the cuticle "inside-out" to expose the CNS.  If the dissection is performed carefully the CNS will remain attached to the cuticle.  We usually keep the CNS attached to the cuticle throughout the fixation and staining steps, and only completely remove the CNS from the cuticle just prior to mounting the samples on glass slides.  We also show some representative images of a larval CNS stained with Eve, a transcription factor expressed in a subset of neurons in the CNS.  The article concludes with a discussion of some of the practical uses of this technique and the potential difficulties that may arise.

In this article we demonstrate how to dissect the central nervous system from third instar Drosophila larvae.

Dissection de SNC larvaire chez Drosophila melanogaster

The central nervous system (CNS) of Drosophila larvae is complex and poorly understood.  One way to investigate the CNS is to use immunohistochemistry to ...

Recherche

Biologie

Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations

Drosophila melanogaster is emerging as a powerful model system for studying the development and function of the nervous system, particularly because of its convenient genetics and fully sequenced genome. Additionally, the larval nervous system is an ideal model system to study mechanisms of axonal transport as the larval segmental nerves contain bundles of axons with their cell bodies located within the brain and their nerve terminals ending along the length of the body. Here we describe the procedure for visualization of synaptic vesicle proteins within larval segmental nerves. If done correctly, all components of the nervous system, along with associated tissues such as muscles and NMJs, remain intact, undamaged, and ready to be visualized. 3rd instar larvae carrying various mutations are dissected, fixed, incubated with synaptic vesicle antibodies, visualized and compared to wild type larvae. This procedure can be adapted for several different synaptic or neuronal antibodies and changes in the distribution of a variety of proteins can be easily observed within larval segmental nerves.

Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations

Drosophila melanogaster larvae provide an ideal model system to investigate the mechanisms of axonal transport within larval segmental nerves. Using this procedure, 3rd instar larvae carrying various mutations can be compared to wild type larvae.

Visualisation des larves nerfs segmentaires en 3 Rd Instar Drosophile Les préparatifs des larves

Drosophila melanogaster is emerging as a powerful model system for studying the development and function of the nervous system, particularly because of ...

Neurosciences

Protocols for Visualizing Steroidogenic Organs and Their Interactive Organs with Immunostaining in the Fruit Fly Drosophila melanogaster

In multicellular organisms, a small group of cells is endowed with a specialized function in their biogenic activity, inducing a systemic response to&#160;growth and reproduction. In insects, the larval prothoracic gland (PG) and the adult female ovary play essential roles in biosynthesizing the principal steroid hormones called ecdysteroids. These ecdysteroidogenic organs are innervated from the nervous system, through&#160;which the timing of biosynthesis is affected by environmental cues. Here we describe a protocol for visualizing ecdysteroidogenic organs and their interactive organs in larvae and adults of the fruit fly Drosophila melanogaster, which provides a suitable model system for studying steroid hormone biosynthesis and its regulatory mechanism. Skillful dissection allows us to maintain the positions of ecdysteroidogenic organs and their interactive organs including the brain, the ventral nerve cord, and other tissues. Immunostaining with antibodies against ecdysteroidogenic enzymes, along with transgenic fluorescence proteins driven by tissue-specific promoters, are available to label ecdysteroidogenic cells. Moreover, the innervations of the ecdysteroidogenic organs can also be labeled by specific antibodies or a collection of GAL4 drivers in various types of neurons. Therefore, the ecdysteroidogenic organs and their neuronal connections can be&#160;visualized simultaneously&#160;by immunostaining and transgenic techniques. Finally, we describe how to visualize germline stem cells, whose proliferation and maintenance are controlled by ecdysteroids. This method contributes to comprehensive understanding of steroid hormone biosynthesis and its neuronal regulatory mechanism.

Protocols for Visualizing Steroidogenic Organs and Their Interactive Organs with Immunostaining in the Fruit Fly Drosophila melanogaster

We describe a protocol for dissection, fixation, and immunostaining of steroidogenic organs in Drosophila larvae and adult females to study steroid hormone biosynthesis and its regulatory mechanism. In addition to steroidogenic organs, we visualize the innervation of steroidogenic organs as well as steroidogenic target cells such as germline stem cells.

Protocoles pour Visualizing stéroïdogénique organes et leurs organes interactifs avec Immunostaining dans le Fruit Fly<em&gt; Drosophila melanogaster</em

In multicellular organisms, a small group of cells is endowed with a specialized function in their biogenic activity, inducing a systemic response ...

Biologie du développement

Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains

Nervous system development involves a sequential series of events that are coordinated by several signaling pathways and regulatory networks. Many of the proteins involved in these pathways are evolutionarily conserved between mammals and other eukaryotes, such as the fruit fly Drosophila melanogaster, suggesting that similar organizing principles exist during the development of these organisms. Importantly, Drosophila has been used extensively to identify cellular and molecular mechanisms regulating processes that are required in mammals including neurogenesis, differentiation, axonal guidance, and synaptogenesis. Flies have also been used successfully to model a variety of human neurodevelopmental diseases. Here we describe a protocol for the step-by-step microdissection, fixation, and immunofluorescent localization of proteins within the adult Drosophila brain. This protocol focuses on two example neuronal populations, mushroom body neurons and retinal photoreceptors, and includes optional steps to trace individual mushroom body neurons using Mosaic Analysis with a Repressible Cell Marker (MARCM) technique. Example data from both wild-type and mutant brains are shown along with a brief description of a scoring criteria for axonal guidance defects. While this protocol highlights two well-established antibodies for investigating the morphology of mushroom body and photoreceptor neurons, other Drosophila brain regions and the localization of proteins within other brain regions can also be investigated using this protocol.

Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains

This protocol describes the dissection and immunostaining of adult Drosophila melanogaster brain tissues. Specifically, this protocol highlights the use of Drosophila mushroom body and photoreceptor neurons as example neuronal subsets that can be accurately used to uncover general principles underlying many aspects of neuronal development.

Dissection et immunofluorescence souillant de champignon corps et photorécepteur de neurones dans le cerveau adulte Drosophila melanogaster

Nervous system development involves a sequential series of events that are coordinated by several signaling pathways and regulatory networks. Many of the ...

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

In this article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the lipophilic fluorescent membrane marker DiI. This method allows the visualization of neuronal cell morphology in great detail. It is possible to label any cell in the CNS: cell bodies of target neurons are visualized under DIC optics or by expression of a fluorescent genetic marker such as GFP. After labeling, the DiI can be transformed into a permanent brown stain by photoconversion to allow visualization of cell morphology with transmitted light and DIC optics. Alternatively, the DiI-labeled cells can be observed directly with confocal microscopy, enabling genetically introduced fluorescent reporter proteins to be colocalised. The technique can be used in any animal, irrespective of genotype, making it possible to analyze mutant phenotypes at single cell resolution.

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

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.

Étiquetage des cellules isolées dans le système nerveux central des<em&gt; Drosophila melanogaster</em

In this  article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the ...

Visualize Drosophila Leg Motor Neuron Axons Through the Adult Cuticle

The majority of work on the neuronal specification has been carried out in genetically and physiologically tractable models such as C. elegans, Drosophila larvae, and fish, which all engage in undulatory movements (like crawling or swimming) as their primary mode of locomotion. However, a more sophisticated understanding of the individual motor neuron (MN) specification&#8212;at least in terms of informing disease therapies&#8212;demands an equally tractable system that better models the complex appendage-based locomotion schemes of vertebrates. The adult Drosophila locomotor system in charge of walking meets all of these criteria with ease, since in this model it is possible to study the specification of a small number of easily distinguished leg MNs (approximately 50 MNs per leg) both using a vast array of powerful genetic tools, and in the physiological context of an appendage-based locomotion scheme. Here we describe a protocol to visualize the leg muscle innervation in an adult fly.

Visualize Drosophila Leg Motor Neuron Axons Through the Adult Cuticle

Here we describe a protocol to visualize the axonal targeting with a florescent protein in adult legs of Drosophila by fixation, mounting, imaging, and post-imaging steps.

Visualiser les Drosophila jambe axones des motoneurones à travers la cuticule de l’adulte

The majority of work on the neuronal specification has been carried out in genetically and physiologically tractable models such as C. elegans, Drosophila ...

Retrograde Tracing of Drosophila Embryonic Motor Neurons Using Lipophilic Fluorescent Dyes

We describe a technique for retrograde labeling of motor neurons in Drosophila. We use an oil-dissolved lipophilic dye and deliver a small droplet to an embryonic fillet preparation by a microinjector. Each motor neuron whose membrane is contacted by the droplet can then be rapidly labeled. Individual motor neurons are continuously labeled, enabling fine structural details to be clearly visualized. Given that lipophilic dyes come in various colors, the technique also provides a means to get adjacent neurons labeled in multicolor. This tracing technique is therefore useful for studying neuronal morphogenesis and synaptic connectivity in the motor neuron system of Drosophila.

Retrograde Tracing of Drosophila Embryonic Motor Neurons Using Lipophilic Fluorescent Dyes

We describe a method for retrograde tracing of the Drosophila embryonic motor neurons using lipophilic fluorescent dyes.

Traçage rétrograde des neurones moteurs embryonnaires drosophila à l'aide de colorants fluorescents lipophiles

We describe a technique for retrograde labeling of motor neurons in Drosophila. We use an oil-dissolved lipophilic dye and deliver a small droplet to an ...

Live Imaging of Drosophila Larval Neuroblasts

Stem cells divide asymmetrically to generate two progeny cells with unequal fate potential: a self-renewing stem cell and a differentiating cell. Given their relevance to development and disease, understanding the mechanisms that govern asymmetric stem cell division has been a robust area of study. Because they are genetically tractable and undergo successive rounds of cell division about once every hour, the stem cells of the Drosophila central nervous system, or neuroblasts, are indispensable models for the study of stem cell division. About 100 neural stem cells are located near the surface of each of the two larval brain lobes, making this model system particularly useful for live imaging microscopy studies. In this work, we review several approaches widely used to visualize stem cell divisions, and we address the relative advantages and disadvantages of those techniques that employ dissociated versus intact brain tissues. We also detail our simplified protocol used to explant whole brains from third instar larvae for live cell imaging and fixed analysis applications.

Live Imaging of Drosophila Larval Neuroblasts

This protocol details a streamlined method used to conduct live cell imaging in the context of an intact larval brain. Live cell imaging approaches are invaluable for the study of asymmetric neural stem cell divisions as well as other neurogenic and developmental processes, consistently uncovering mechanisms that were previously overlooked.

Imagerie en direct de<em&gt; Drosophila</em&gt; larves Neuroblastes

Stem cells divide asymmetrically to generate two progeny cells with unequal fate potential: a self-renewing stem cell and a differentiating cell. Given ...

Dissection, Immunohistochemistry and Mounting of Larval and Adult Drosophila Brains for Optic Lobe Visualization

The Drosophila optic lobe, comprised of four neuropils: the lamina, medulla, lobula and lobula plate, is an excellent model system for exploring the developmental mechanisms that generate neural diversity and drive circuit assembly. Given its complex three-dimensional organization, analysis of the optic lobe requires that one understand how its adult neuropils and larval progenitors are positioned relative to each other and the central brain. Here, we describe a protocol for the dissection, immunostaining and mounting of larval and adult brains for optic lobe imaging. Special emphasis is placed on the relationship between mounting orientation and the spatial organization of the optic lobe. We describe three mounting strategies in the larva (anterior, posterior and lateral) and three in the adult (anterior, posterior and horizontal), each of which provide an ideal imaging angle for a distinct optic lobe structure.

Dissection, Immunohistochemistry and Mounting of Larval and Adult Drosophila Brains for Optic Lobe Visualization

This protocol describes three steps to prepare larval and adult Drosophila optic lobes for imaging: 1) brain dissections, 2) immunohistochemistry and 3) mounting. Emphasis is placed on step 3, as distinct mounting orientations are required to visualize specific optic lobe structures.&#160;

Dissection, immunohistochimie et montage de cerveaux de drosophiles larvaires et adultes pour la visualisation du lobe optique

The Drosophila optic lobe, comprised of four neuropils: the lamina, medulla, lobula and lobula plate, is an excellent model system for exploring the ...

Upright Imaging of Drosophila Embryos

Several well-known morphogenetic gradients and cellular movements occur along the dorsal/ventral axis of the Drosophila embryo. However, the current techniques used to view such processes are somewhat limited. The following protocol describes a new technique for mounting fixed and labeled Drosophila embryos for coronal viewing with confocal imaging. This method consists of embedding embryos between two layers of glycerin jelly mounting media, and imaging jelly strips positioned upright. The first step for sandwiching the embryos is to make a thin bedding of glycerin jelly on a slide. Next, embryos are carefully aligned on this surface and covered with a second layer of jelly. After the second layer is solidified, strips of jelly are cut and flipped upright for imaging. Alternatives are described for visualizing the embryos depending upon the type of microscope stand to be used. Since all cells along the dorsal-ventral axis are imaged within a single confocal Z-plane, our method allows precise measurement and comparison of fluorescent signals without photobleaching or light scattering common to 3D reconstructions of longitudinally mounted embryos.

Upright Imaging of Drosophila Embryos

Here we present a mounting protocol for stained Drosophila embryos in an upright position that allows imaging of cross-sections using Confocal microscopy.

Imagerie verticale de<em&gt; Drosophila</em&gt; Embryons

Several well-known morphogenetic gradients and cellular movements occur along the dorsal/ventral axis of the Drosophila embryo. However, the current ...

La visualisation du système nerveux embryonnaire, en totalité montage Drosophile Embryons

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Dissection de SNC larvaire chez Drosophila melanogaster

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Protocoles pour Visualizing stéroïdogénique organes et leurs organes interactifs avec Immunostaining dans le Fruit Fly

Dissection et immunofluorescence souillant de champignon corps et photorécepteur de neurones dans le cerveau adulte Drosophila melanogaster

Étiquetage des cellules isolées dans le système nerveux central des

Visualiser les Drosophila jambe axones des motoneurones à travers la cuticule de l’adulte

Traçage rétrograde des neurones moteurs embryonnaires drosophila à l'aide de colorants fluorescents lipophiles

Imagerie en direct de

Dissection, immunohistochimie et montage de cerveaux de drosophiles larvaires et adultes pour la visualisation du lobe optique

Imagerie verticale de