Name: Twee-foton laser-gemedieerde ablatie van osteoblasten in zich ontwikkelende zebravislarven (Video) | JoVE Journal
Uploaded: 2024-09-06T14:50:00.000+00:00
Duration: 5 min 46 s
Description: Leer twee-foton laserablatie van osteoblasten in zich ontwikkelende Danio rerio larven. Deze methode onderzoekt botregeneratie en de rol van het immuunsysteem bij het herstelproces.

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<meta charset="utf8"></head><body>Laserablatiemethode om botregeneratie bij zebravislarven te bestuderen

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<table><tbody><tr><td>Calcium chloride dihydrate, CaCl2&middot;H2O</td><td>Carl Roth</td><td>5239.1</td><td></td></tr><tr><td>Cell Culture Dish, PS, 100/20 mm</td><td>Greiner Bio-one</td><td>664160</td><td></td></tr><tr><td>Dumont #55 Foceps</td><td>FST</td><td>11295-51</td><td>Tip shape straight, 11 cm, 0.05 x 0.02 mm</td></tr><tr><td>Falcon 6-well plate</td><td>Corning</td><td>353502</td><td></td></tr><tr><td>Glass-bottom microwell dish</td><td>MatTek</td><td>P35G-1.5-14-C</td><td>35 mm Dish, No. 1.5 Coverslip, 14 mm Glass Diameter, Uncoated</td></tr><tr><td>Insight X3 multiphoton laser</td><td>Spectra-Physics</td><td></td><td></td></tr><tr><td>Leica Application Suite</td><td>LAS X, Leica Microsystems</td><td></td><td></td></tr><tr><td>Low melting agarose&nbsp;</td><td>Biozyme</td><td>840101</td><td>Biozym Plaque Agarose</td></tr><tr><td>Magnesium sulfate heptahydrate, MgSO4&middot;7H2O</td><td>Sigma-Aldrich</td><td>M5921</td><td></td></tr><tr><td>Mating cages</td><td></td><td></td><td>many varieties, e.g. Tecniplast</td></tr><tr><td>Methyleneblue</td><td>Carl Roth</td><td>A514.1</td><td></td></tr><tr><td>MS-222</td><td>SIGMA Aldrich</td><td>A5040</td><td></td></tr><tr><td>Potassium Chloride, KCl</td><td>PanReac AppliChem</td><td>131494</td><td></td></tr><tr><td>Sodium chloride, NaCl</td><td>Carl Roth</td><td>3957.1</td><td></td></tr><tr><td>SP8 FALCON&nbsp;</td><td>Leica Microsystems</td><td></td><td>Equipped with a Insight X3 multiphoton laser and Leica Application Suite software</td></tr><tr><td>Stainless Steel Dissect Needle</td><td>Bochem</td><td>12010</td><td>140 mm</td></tr><tr><td>Stereo Microscope System SZX16</td><td>Olympus</td><td></td><td>Equipped with a LED illumination base SZX2-ILLTQ</td></tr><tr><td>Thermostatic Cabinets TS - WTW</td><td>xylem</td><td>TS 608/2-i</td><td>For incubation (embryo)</td></tr><tr><td>Transfer pipette, 3.5 mL</td><td>SARSTEDT</td><td>861171</td><td>155 x 15 mm, LD-PE, transparent</td></tr><tr><td>Zeiss SteREO Discovery.V12 version 4.7.1.0.</td><td>Zeiss</td><td></td><td>Equipped with Axiocam MRm camera and AxioVision sofware</td></tr></tbody></table>

twee-foton laser-gemedieerde ablatie van osteoblasten in zich ontwikkelende zebravislarven

Two-Photon Laser-Mediated Ablation of Osteoblasts in Developing Zebrafish Larvae

Twee-foton laser-gemedieerde ablatie van osteoblasten in zich ontwikkelende zebravislarven

Zebrafish (Danio rerio) have an outstanding capacity to regenerate different organs and appendages. Bone regeneration in zebrafish has been studied using ...

two-photon-laser-mediated-ablation-osteoblasts-developing-zebrafish

Research

JoVE Journal

Developmental Biology

Hier beschrijven we een methode voor het monteren van zebravis embryo's voor de lange termijn imaging, twee-foton beeldvorming en weefsel-schade technieken en time-lapse confocale beeldvorming.

Twee-foton axotomy en time-lapse confocale imaging in levende zebravis embryo&#39;s

Acute nierschade (AKI) bij de mens is een veel voorkomend klinisch probleem wordt veroorzaakt door schade aan de epitheliale cellen die de nieren nefronen bevatten, en de AKI wordt geassocieerd met een hoge sterfte van...

Laser Ablatie van de zebravis Pronephros renale epitheliale regeneratie Study

De werkwijze omvat de hier gepresenteerde precieze schade van levende zebravis embryo's met hoge energie laserpulsen en de analyse van deze letsels en hun herstel met de tijd. We laten ook zien hoe genetisch enkel label of groepen van skeletspiercellen kunnen worden gevolgd tijdens en na de laser-licht veroorzaakte schade.

Laser toegebrachte schade van zebravis Embryonale skeletspier

Deze video beschrijft de methode om axonen in zebravisembryo's te verwonden met behulp van twee fotonlaseraxotomie en het observeren van axonale herstel van letsel.

Twee-Foton Laser Axotomie: Een methode om axonen in zebravisembryo's te verwonden en axonale herstel te observeren

Source: Morsch, M.,  et. al., Triggering Cell Stress and Death Using Conventional UV Laser Confocal Microscopy. J. Vis. Exp. (2017) In this video, we explore the technique of high-intensity UV laser ablation for precisely targeting and studying individual neurons in transgenic zebrafish. We demonstrate how to prepare, immobilize, and visualize neuronal responses, highlighting the cellular dynamics of injury and recovery.

Selective Single-Neuron Laser Ablation in a Zebrafish Larva Using a Confocal Microscope

Source: Muto, A., et al. Ablation of a Neuronal Population Using a Two-photon Laser and Its Assessment Using Calcium Imaging and Behavioral Recording in Zebrafish Larvae. J. Vis. Exp. (2018) In this video, a two-photon laser ablation procedure is demonstrated, enabling the selective destruction of fluorescent neurons in agarose-immobilized zebrafish larvae. High-resolution imaging before and after the procedure confirms efficient neuronal damage, as observed by the absence of fluorescence.

Targeted Neuronal Ablation Using Two-Photon Microscopy in a Zebrafish Larava

Hier presenteren we een protocol om een genetisch gelabelde subpopulatie van neuronen door een twee-foton laser vanaf Zebrafish larven lasertherapie.

Ablatie van een neuronale bevolking met behulp van een twee-foton Laser en haar beoordeling met Calcium Imaging en gedrags opname in Zebrafish larven

Laserablatie is een breed toepasbare technologie voor het bestuderen van regeneratie in biologische systemen. Het gepresenteerde protocol beschrijft het gebruik van een standaard laserscanning confocale microscoop voor laserablatie en de daaropvolgende time-lapse beeldvorming van het regenereren van interneuromastcellen in de zebravis laterale lijn.

Confocale Microscoop-gebaseerde Laser Ablatie en Regeneratie Test in Zebrafish Interneuromast Cellen

Embryonale ontwikkeling vereist grootschalige coördinatie van celbewegingen. Twee-foton excitatie gemedieerde laserablatie maakt de ruimtelijk gecontroleerde 3-dimensionale ablatie van grote groepen diepe cellen mogelijk. Bovendien kan deze techniek de reactie van collectief migrerende cellen in vivo op verstoringen in hun mechanische omgeving onderzoeken.

Diepe en ruimtelijk gecontroleerde volume-ablaties met behulp van een twee-fotonenmicroscoop in de zebravis Gastrula

We presenteren een protocol voor het vastleggen van de dynamica van regeneratie van de staartvin van zebravislarven op een volledige weefselschaal met behulp van stereomicroscopie op basis van helder veld. Deze techniek maakt het mogelijk om de regeneratiedynamica met een enkele celresolutie vast te leggen. Deze methodologie kan worden aangepast aan elke stereomicroscoop uitgerust met een CCD-camera en time-lapse software.

Vastleggen van Tissue Repair in zebravis Larven met Time-lapse Helderveld stereomicroscopie

Leer twee-foton laserablatie van osteoblasten in zich ontwikkelende Danio rerio larven. Deze methode onderzoekt botregeneratie en de rol van het immuunsysteem bij het herstelproces.

Twee-foton laser-gemedieerde ablatie van osteoblasten in zich ontwikkelende zebravislarven

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