Name: Onderzoek naar retinale circuits en moleculaire lokalisatie door pre-embedding immuno-elektronenmicroscopie (Video) | JoVE Journal
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Description: Verken pre-embedding immuno-elektronenmicroscopie voor retinale circuits en neurotransmitterlokalisatie in muizenretina. Deze methode onthult synaptische interconnectiviteit tussen kegel- en staafbanen.

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<meta charset="utf8"></head><body>Analyse van synaptische circuits en eiwitlokalisatie in het netvlies van muizen

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onderzoek naar retinale circuits en moleculaire lokalisatie door pre-embedding immuno-elektronenmicroscopie

Investigating Retinal Circuits and Molecular Localization by Pre&#45;Embedding Immunoelectron Microscopy

Onderzoek naar retinale circuits en moleculaire lokalisatie door pre-embedding immuno-elektronenmicroscopie

The retina comprises numerous cells forming diverse neuronal circuits, which constitute the first stage of the visual pathway. Each circuit is ...

investigating-retinal-circuits-molecular-localization-pre-embedding

Research

JoVE Journal

Neuroscience

Neurowetenschap

De neonatale muizen netvlies biedt een goed gekarakteriseerd fysiologisch model van angiogenese, die onderzoeken van de rol van de verschillende genen of medicijnen die angiogenese moduleren in een...

Hele Mount immunofluorescentiekleuring van de Neonatale Mouse Retina naar Angiogenese Onderzoek<em&gt; In vivo</em

Het hier beschreven protocol is voor structurele beoordeling van een wholemount retinale preparatie. Dit omvat beschrijvingen van weefseldissecatie, montage op een hydrofiliseerde polytetrafluorethyleen (PTFE) membraaninzet, bolusbelasting met fluorescente markeringen, en een vergelijking van fixatie met carbodiimide en paraformaldehyde voor immunohistochemische analyse van cellulaire en synaptische componenten.

Geoptimaliseerd protocol voor Retina Wholemount Voorbereiding voor Imaging en Immunohistochemistry

De postembedding immunogold-methode is een van de meest effectieve manieren om hoogwaardige analyses van de subcellulaire lokalisatie van specifieke moleculen te leveren. Hier beschrijven we een protocol om glutamaatreceptoren bij retinale lintsynapsen kwantitatief te analyseren.

High-Resolution Kwantitatieve immunogoud Analyse van membraanreceptoren op het netvlies Ribbon Synapses

We tonen drie verschillende weefsel voorbereiding technieken voor immunohistochemische visualisatie van rat retinale microvasculaire pericytes, dat wil zeggen, cryo-secties, geheel-mounts en hypotone isolatie van de vasculaire netwerk.

Netvlies Cryo-secties, geheel-Mounts en hypotone geïsoleerde therapieën voorbereidingen voor immunohistochemische visualisatie van microvasculaire Pericytes

Dit rapport beschrijft uitgebreide methoden voor de voorbereiding van bevroren muis Retina secties voor Immunohistochemistry (IHC).  Beschreven methoden omvatten dissectie van de oculaire posterior Cup, Paraformaldehyde fixatie, inbedding in optimale snij-temperatuur (OCT) media en weefsel oriëntatie, sectioneren en immunokleuring.

Voorbereiding van de muis netvlies Cryo-profielen voor Immunohistochemistry

Methanol kan worden gebruikt als een hulpmedium voor retinale preparaten voor de gehele montage van het netvlies en langdurige opslag, wat nuttig is voor het onderzoek van retinale ganglioncellen.

Op methanol gebaseerd hele-mountpreparaat voor het onderzoek van retinale ganglioncellen

Dit protocol beschrijft de gedetailleerde stappen voor het voorbereiden van netvliesmonsters voor volume-elektronenmicroscopie, met de nadruk op de structurele kenmerken van retinale fotoreceptorterminals.

Voorbereiding van netvliesmonsters voor volume-elektronenmicroscopie

Dit protocol biedt een geoptimaliseerde en uitgebreide voorbereidingsprocedure voor retinale organoïdemonsters voor transmissie-elektronenmicroscopie. Het is geschikt voor toepassingen waarbij synapsen in volwassen retinale organoïden worden geanalyseerd.

Voorbereiding van retinale organoïdemonsters voor transmissie-elektronenmicroscopie

Er wordt een protocol beschreven voor het voorbereiden van retinale cryosecties van muizen en het uitvoeren van immunokleuring op fotoreceptoren. Dit artikel stelt onderzoekers in staat om consequent bevroren secties van het netvlies van muizen te produceren met goed bewaarde morfologie en hoogwaardige immunokleuringsresultaten.

Cryosectie en immunokleuring van het netvlies van de muis

Het netvlies vertoont een complexe ruimtelijke organisatie over zijn laminaire structuur. Moderne ruimtelijke transcriptomics en sequencingtechnieken vereisen weefselcoupes &lt; 20 μm dik, waardoor hun toepasbaarheid voor retinaal onderzoek wordt beperkt. Deze vers ingevroren cryosectiemethode produceert dunne delen van het netvlies van de muis met behoud van vlakke ruimtelijke relaties, waardoor uitgebreide moleculaire kartering over laminae mogelijk is.

En gezicht Cryosectie van het netvlies van de muis voor hoogdimensionale ruimtelijke moleculaire analyse

Verken pre-embedding immuno-elektronenmicroscopie voor retinale circuits en neurotransmitterlokalisatie in muizenretina. Deze methode onthult synaptische interconnectiviteit tussen kegel- en staafbanen.

Onderzoek naar retinale circuits en moleculaire lokalisatie door pre-embedding immuno-elektronenmicroscopie

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