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…

Açıklamalar

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
Pipettor tips [2-200 μL, 50-1,000 μL] Eppendorf epT.I.P.S.
22491334;
022491351
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

Etiketler

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Bu Makaleden Alıntı Yapın
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).

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