Injection of Microbubbles into Isolated Mouse Embryos: A Technique to Deliver Microbubble Contrast Agents into the Vasculature of Living Murine Embryos
Published: April 30, 2023
Abstract
Source: Denbeigh, J. M. et al. Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound. J. Vis. Exp. (2015)
This video demonstrates a procedure to inject microbubble contrast agents into an isolated late gestation stage mouse embryo. The microbubble contrast agent-based ultrasound imaging helps visualize and characterize the vascular environment of the embryo.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Microbubble Preparation Preparing microbubbles for injection NOTE: The assistant performs this step when ‘Injection of Microbubbles (MB) into Embryos’ is commenced. Each embryo is injected once, with the type of microbubble chosen for each injection to be randomly selected by the assi…
Divulgazioni
The authors have nothing to disclose.
Materials
| Ice | |||
| 0.9% sodium chloride (saline) | Hospira | 0409-7984-11 | |
| Phosphate Buffered Saline [1x] | Sigma | D8537 | 1x, w/o calcium chloride & magnesium chloride |
| Ultrasound contrast agent, target ready and untargeted | MicroMarker; VisualSonics Inc. | ||
| Ultrasound gel (Aquasonic 100, colourless) | CSP Medical | 133-1009 | |
| Ice bucket | Cole Parmer | RK 06274-01 | |
| Imaging Platform | VisualSonics Inc. | Integrated Rail System | |
| Light source, fiber-optic | Fisher Scientific | 12-562-36 | Ideally has adjustable arms |
| Luers (12), polypropylene barbed female ¼-28 UNF thread | Cole Parmer | 45500-30 | |
| Micro-ultrasound system, high-frequency | VisualSonics Inc. | Vevo2100 | |
| Needles, 21 gauge (1”) | VWR | 305165 | |
| Perforated spoon (Moria) | Fine Science Tools | MC 17 10373-17 | |
| Pins (6), black anodized minutien 0.15 mm | Fine Science Tools | 26002-15 | |
| Syringes, 1 mL Normject | Fisher | 14-817-25 | |
| Syringe infusion pump | Bio-lynx | NE-1000 | |
| Pipettors [2-20 μL, 20-200 μL, 100-1,000 μL] | Eppendorf | Research Plus adjustable 3120000038; 3120000054; 3120000062 |
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| Pipettor tips [2-200 μL, 50-1,000 μL] | Eppendorf | epT.I.P.S. 22491334; 022491351 |
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| Scissors | |||
| Glass capillaries, 1 x 90 mm GD-1 with filament | Narishige | GD-1 | |
| Glass needle puller | Narishige | PN-30 | |
| Tubes, Eppendorf | VWR | 20170-577 | |
| Tube racks (3) | VWR | 82024-462 | |
| Hot plate | VWR | 89090-994 | |
| Syringes (10), 30 mL | VWR | CA64000-041 | |
| Ultrasound transducer, 20 MHz | VisualSonics Inc. | MS250 |
Tags
Citazione di questo articolo
Injection of Microbubbles into Isolated Mouse Embryos: A Technique to Deliver Microbubble Contrast Agents into the Vasculature of Living Murine Embryos. J. Vis. Exp. (Pending Publication), e20743, doi: (2023).