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Bioengineering
Schnelle hochauflösende Bildgebung des Gefäßsystems von Kleintieren für quantitative Studien
Schnelle hochauflösende Bildgebung des Gefäßsystems von Kleintieren für quantitative Studien
JoVE Journal
Bioengineering
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JoVE Journal Bioengineering
Rapid Whole-Mount High-Resolution Imaging of Small Animal Vasculature for Quantitative Studies

Schnelle hochauflösende Bildgebung des Gefäßsystems von Kleintieren für quantitative Studien

Full Text
821 Views
08:49 min
May 23, 2025

DOI: 10.3791/68206-v

Daibo Zhang1, Stephanie E. Lindsey1

1Mechanical and Aerospace Engineering,University of California San Diego

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This protocol introduces a rapid method for quantitative whole-mount three-dimensional vascular imaging using light-sheet fluorescence microscopy. The technique is demonstrated using the pharyngeal arch artery system of the chick embryo model, allowing for the quantification of hemodynamic forces through computational fluid dynamics.

Key Study Components

Area of Science

  • Neuroscience
  • Vascular Imaging
  • Embryology

Background

  • Understanding mechanisms of disease formation, particularly congenital heart defects.
  • Investigating the role of hemodynamic flow in disease development.
  • Utilizing various imaging techniques for high-resolution scans.
  • Addressing the need for accessible vascular imaging methods.

Purpose of Study

  • To provide a rapid and efficient vascular imaging technique.
  • To apply the method across various animal models, including embryos.
  • To reduce sample preparation time significantly.

Methods Used

  • Light-sheet fluorescence microscopy for imaging.
  • Computational fluid dynamics for hemodynamic analysis.
  • Minimal use of hazardous clearing compounds.
  • Application of the method to chick embryo models.

Main Results

  • Demonstrated efficacy of the imaging method on the pharyngeal arch artery system.
  • Quantified hemodynamic forces effectively.
  • Reduced sample preparation time from four weeks to three days.
  • Enabled imaging without reliance on vascular markers in early embryos.

Conclusions

  • The protocol offers a time-efficient solution for vascular imaging.
  • It is applicable to various animal models and their developmental stages.
  • The method enhances the understanding of vascular development and disease mechanisms.

Frequently Asked Questions

What is the main advantage of this imaging technique?
The main advantage is its rapid sample preparation time, reducing it from four weeks to just three days.
Can this method be applied to other animal models?
Yes, the protocol is designed to be applicable to various animal models, including embryos.
What imaging techniques are compared in this study?
The study compares light-sheet fluorescence microscopy with other imaging techniques like nano-computed tomography and 4D ultrasound.
How does this method contribute to understanding congenital heart defects?
It allows for the quantification of hemodynamic forces, which may contribute to the formation of congenital heart defects.
What are the implications of using minimal hazardous compounds?
Using minimal hazardous compounds makes the method safer and more accessible for use in various research settings.

Dieses Protokoll stellt eine schnelle Methode für die quantitative dreidimensionale Gefäßbildgebung mit Hilfe der Lichtblatt-Fluoreszenzmikroskopie vor. Die Wirksamkeit der Methode wird anhand des pharyngealen Bogenarteriensystems des Hühnerembryomodells demonstriert, wobei die hämodynamischen Kräfte mittels Computational Fluid Dynamics quantifiziert werden.

Wir arbeiten daran, die Mechanismen der Krankheitsentstehung zu verstehen, insbesondere im Zusammenhang mit angeborenen Herzfehlern und ob Veränderungen des hämodynamischen Flusses zur Krankheitsentstehung beitragen oder diese verstärken. Nano-Computertomographie, Magnetresonanztomographie bei kleinen Tieren, 4D-Ultraschall und optische Kohärenzdemografie werden alle verwendet, um hochauflösende, quantitative Bildgebungsscans für die Computermodellierung zu sammeln.

Unser Protokoll adressiert den Bedarf an einer schnellen und leicht zugänglichen quantitativen Gefäßbildgebungstechnik, die auf eine Vielzahl von Tiermodellen, einschließlich ihrer Embryonen und Organe, angewendet werden kann. Unser Protokoll ist zeiteffizient und verkürzt die Probenvorbereitungszeit von vier Wochen auf nur drei Tage. Es verwendet minimal gefährliche Clearing-Verbindungen und verlässt sich nicht auf Gefäßmarker, die in frühen Embryonen nicht vorhanden sind.

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