Name: In-vivo en ex-vivo calciumbeeldvorming van een reukcircuitneuron in de larve van de Drosophila melanogaster met een derde instar (Video) | JoVE Journal
Uploaded: 2025-10-31T14:55:00.000+00:00
Duration: 11 min 58 s
Description: Leer calciumbeeldvorming in reukcircuitneuronen van Drosophila melanogaster-larven met GLUture-kleefmiddel. Deze methode biedt stabiele in-vivo en ex-vivo opstellingen voor betrouwbare neurofysiologische experimenten.

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<table><tbody><tr><td>Fly Stocks &amp; Onderhoud</td><td></td><td></td><td></td></tr><tr><td>Nutrifly vliegvoer</td><td>Genesee Scientific</td><td>66-112</td><td></td></tr><tr><td>UAS-GCaMP6f, tdTomato</td><td>Gift van Dr. JH Kim&nbsp;</td><td></td><td></td></tr><tr><td>w;;Keystone-Gal4 (GMR27F08-GAL4)</td><td>BDSC</td><td>49232</td><td></td></tr><tr><td>Reagentia voor hongerprotocol</td><td></td><td></td><td></td></tr><tr><td>6 cm Petri schotel</td><td>Genesee Scientific</td><td>32-106G</td><td></td></tr><tr><td>Sucrose</td><td>Millipore Sigma</td><td>S0389-500G</td><td></td></tr><tr><td>Reagentia voor beeldvorming en dissectiebuffer</td><td></td><td></td><td></td></tr><tr><td>Calciumchloride</td><td>J.T. Baker, Avantor Sciences</td><td>JT1311-1</td><td>CAS: 1311-01, 500g</td></tr><tr><td>D-(+)-Glucose</td><td>Thermo Fisher Scientific</td><td>A16828.36</td><td>CAS: 50-99-7, 500g</td></tr><tr><td>HEPES-2-[4-(2-Hydroxyethyl)-1-piperazinyl]-ethaansulfonzuur</td><td>J.T. Baker, Avantor Sciences</td><td>JT4018-1</td><td>CAS: 4018-01, 100g</td></tr><tr><td>Magnesiumchloride</td><td>VWR</td><td>97064-150</td><td>CAS: 7786-30-3, 100g</td></tr><tr><td>Kaliumchloride</td><td>Thermo Fisher Scientific</td><td>J64189.36</td><td>CAS: 7447-40-7, 500g</td></tr><tr><td>Natriumchloride</td><td>EMD Milipore Sigma</td><td>EMD-SX0420-1</td><td>CAS: 7647-14-5, 500g</td></tr><tr><td>Hulpmiddelen en Reagentia voor Dissectie</td><td></td><td></td><td></td></tr><tr><td>Dissectiebuffer</td><td></td><td></td><td>Samenstelling vermeld in tekst</td></tr><tr><td>Olympus Petri schotels, 60 x 15mm Geventileerd</td><td>Genesee Scientific</td><td>32-105</td><td></td></tr><tr><td>Sylgard 184 Elastomer Basis</td><td>Electron Microscopy Sciences</td><td>24236-10A</td><td></td></tr><tr><td>Sylgard 184 Elastomer Basis Uithardingsmiddel</td><td>Electron Microscopy Sciences</td><td>24236-10B</td><td></td></tr><tr><td>Pincet Stijl 5, EMS</td><td>Electron Microscopy Sciences</td><td>78320-5</td><td></td></tr><tr><td>Vannas Lenteschaart - 3mm Snijrand</td><td>Fine Science Tools</td><td>15000-00</td><td></td></tr><tr><td>Beeldvorming en Montageapparatuur/Reagentia</td><td></td><td></td><td></td></tr><tr><td>Confocal Scanner module</td><td>Yokogawa</td><td>CSU-W1</td><td>Aangesloten op microscoop</td></tr><tr><td>Beeldvorming Buffer</td><td></td><td></td><td>Samenstelling vermeld in tekst</td></tr><tr><td>Omgekeerde Microscoop</td><td>Leica Microsystems</td><td>DMi8&nbsp;</td><td>Aangesloten op een spinning disc confocale scanner module en een CCD-camera</td></tr><tr><td>Microscoop Dekglas, 24mm x 50mm, # 1.5 Dikte</td><td>Globe Scientific Inc.</td><td>1415-15</td><td></td></tr><tr><td>MWI Veterinary GLUTURE Topisch Adhesief</td><td>Fisher Scientific</td><td>NC0632797</td><td></td></tr><tr><td>Pure Petroleum Jelly</td><td>Walgreens</td><td>912556</td><td></td></tr><tr><td>Links naar Openbaar Beschikbare Bronnen</td><td></td><td></td><td></td></tr><tr><td>BioRender</td><td></td><td></td><td>https://www.biorender.com/</td></tr><tr><td>ImageJ</td><td>NIH</td><td>912556</td><td>https://imagej.net/ij/</td></tr><tr><td>Janelia Flylight protocol video</td><td>https://www.janelia.org/</td><td></td><td>https://www.youtube.com/watch?v=UVCPmuu2b6o</td></tr><tr><td>Mathew Lab Github</td><td></td><td></td><td>https://github.com/MathewLab/JoVE-Calcium-imaging</td></tr></tbody></table>

in-vivo en ex-vivo calciumbeeldvorming van een reukcircuitneuron in de larve van de drosophila melanogaster met een derde instar

In-vivo and Ex-vivo Calcium Imaging of an Olfactory Circuit Neuron in the Third-Instar Drosophila melanogaster Larva

In-vivo en ex-vivo calciumbeeldvorming van een reukcircuitneuron in de larve van de Drosophila melanogaster met een derde instar

Calcium imaging is crucial for studying neural dynamics and brain function. However, current methods for imaging calcium in Drosophila larvae are ...

in-vivo-ex-vivo-calcium-imaging-an-olfactory-circuit-neuron-third

Research

JoVE Journal

Neuroscience

Neurowetenschap

Dit protocol beschrijft een betrouwbare methode voor verdoving en beeldvorming van...

In vivo beeldvorming van Intact Drosophila larven aan Subcellulaire resolutie

We beschrijven een gevestigde techniek om te meten en geur-opgeroepen calcium antwoorden te analyseren in de antennaal kwab van het...

Calcium Imaging van Geur-reacties van de consument in de Drosophila Antennaal Lobe

Deze video beschrijft in vivo calcium beeldvorming in Drosophila neuronen met behulp van GCaMP. De aanbevolen protocolclip laat zien hoe u veranderingen in GCaMP-fluorescentie van paddestoellichaamneuronen kunt registreren tijdens een olfactorisch conditioneringsexperiment.

Drosophila In Vivo Calcium imaging: een methode voor functionele beeldvorming van neuronale activiteit

This video demonstrates a method for in vivo calcium imaging using synaptically localized calcium sensors in flies. It outlines the steps for preparing the flies for microscopy, conducting olfactory conditioning, and visualizing odor-induced calcium influx.

In Vivo Calcium Imaging Using Synaptically Localized Calcium Sensors in Drosophila melanogaster

Source: Ishimoto, H. et. al., Ex Vivo Calcium Imaging for Visualizing Brain Responses to Endocrine Signaling in Drosophila. J. Vis. Exp. (2018) This video demonstrates calcium imaging to study Drosophila brain responses to neuropeptides. Neurons expressing a fluorescent calcium indicator reveal intracellular calcium dynamics and functional responses to signaling molecules.

Ex Vivo Calcium Imaging to Study Drosophila Brain Responses to Neuropeptides

Hier we illustreren hoe enkel molecuul foto-geactiveerde lokalisatie microscopie kan worden uitgevoerd op de motorische zenuw-terminal van een levende Drosophila melanogaster derde instar larve.

In Vivo Single-molecuul volgen bij de Terminal van de presynaptische motorische zenuw Drosophila

Dit witboek beschrijft een protocol voor ex vivo calcium beeldvorming van de hersenen van Drosophila . Bij deze methode, kunnen natuurlijke of synthetische verbindingen worden toegepast op de buffer voor het testen van hun vermogen om bepaalde neuronen in de hersenen activeren.

Ex Vivo Calcium Imaging voor het visualiseren van hersenen reacties op endocriene signalering in Drosophila

Hier presenteren we een protocol waarmee pre-en/of postsynaptisch calcium kan worden gevisualiseerd in de context van Drosophila leren en geheugen. In vivo calcium beeldvorming met behulp van Synaptisch gelokaliseerde calcium sensoren wordt gecombineerd met een klassieke olfactorische conditionerings paradigma zodanig dat de Synaptische plasticiteit onderliggende dit type associatief leren kan worden bepaald.

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Hier bespreken we een workflow om levende explant-hersenen van Drosophila melanogaster derde instarlarven voor te bereiden, te ontleden, te monteren en in beeld te brengen om de cellulaire en subcellulaire dynamiek onder fysiologische omstandigheden te observeren.

Live-Cell Imaging van Drosophila melanogaster Derde Instar Larvale Hersenen

Hier presenteren we een protocol voor ex vivo calciumbeeldvorming bij volwassen Drosophila met GCaMP6-expressie om epileptiforme activiteiten te monitoren. Het protocol biedt een waardevol hulpmiddel voor het onderzoeken van ictale gebeurtenissen bij volwassen Drosophila door middel van ex vivo calciumbeeldvorming, waardoor de mogelijke mechanismen van epilepsie op cellulair niveau kunnen worden onderzocht.

Ex Vivo Calciumbeeldvorming voor Drosophila-model van epilepsie

Een calciumbeeldvormingsbenadering maakt het mogelijk om smaakgeïnduceerde reacties in de hersenen van wakkere vliegen vast te leggen terwijl een oplossing op het labellum wordt aangebracht. Primaire smaakresponsen in Drosophila melanogaster worden als voorbeeld gebruikt, maar dit protocol kan worden aangepast om stroomafwaartse neuronen of andere soorten te bestuderen.

In vivo Calciumbeeldvorming van smaakgeïnduceerde neurale reacties bij volwassen Drosophila

Leer calciumbeeldvorming in reukcircuitneuronen van Drosophila melanogaster-larven met GLUture-kleefmiddel. Deze methode biedt stabiele in-vivo en ex-vivo opstellingen voor betrouwbare neurofysiologische experimenten.

In-vivo en ex-vivo calciumbeeldvorming van een reukcircuitneuron in de larve van de Drosophila melanogaster met een derde instar (Video) | JoVE Journal

In-vivo en ex-vivo calciumbeeldvorming van een reukcircuitneuron in de larve van de Drosophila melanogaster met een derde instar

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