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Medicine
염화제이철 유발 동맥 혈전증 및 3D 전자 현미경 분석을 위한 시료 수집
염화제이철 유발 동맥 혈전증 및 3D 전자 현미경 분석을 위한 시료 수집
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Medicine
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JoVE Journal Medicine
Ferric Chloride-Induced Arterial Thrombosis and Sample Collection for 3D Electron Microscopy Analysis

염화제이철 유발 동맥 혈전증 및 3D 전자 현미경 분석을 위한 시료 수집

Full Text
2,502 Views
03:57 min
March 17, 2023

DOI: 10.3791/64985-v

Smita Joshi*1, Alexis N. Smith1, Kanakanagavalli Shravani Prakhya1, Hammodah R. Alfar1, Joshua Lykins1, Ming Zhang1, Irina Pokrovskaya2, Maria Aronova3, Richard D. Leapman3, Brian Storrie2, Sidney W. Whiteheart*1

1Department of Molecular and Cellular Biochemistry,University of Kentucky, 2Department of Physiology and Cell Biology,University of Arkansas for Medical Sciences, 3Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering,National Institutes of Health

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Overview

This protocol outlines a FeCl3-mediated method to induce arterial thrombosis and collect samples for electron microscopy analysis. It allows researchers to study the structural aspects of thrombus growth and architecture at various stages.

Key Study Components

Area of Science

  • Neuroscience
  • Thrombosis
  • Hemostasis

Background

  • Understanding thrombosis is crucial for developing treatments for cardiovascular diseases.
  • Platelet and hemostatic system components play significant roles in thrombus formation.
  • Electron microscopy provides detailed structural insights into thrombus development.
  • Reproducibility in experimental methods is essential for reliable results.

Purpose of Study

  • To identify key components involved in thrombosis.
  • To analyze how thrombus growth is influenced at the cellular level.
  • To establish a reproducible method for studying thrombosis stages.

Methods Used

  • FeCl3-mediated arterial injury induction.
  • Sample collection at various thrombosis stages.
  • Preparation of samples for electron microscopy.
  • Careful handling of tissue to avoid complications.

Main Results

  • Identification of structural changes in thrombus over time.
  • Insights into the cellular mechanisms driving thrombus formation.
  • Establishment of a reliable protocol for future studies.
  • Recommendations for minimizing tissue handling during procedures.

Conclusions

  • The method provides a reproducible framework for studying thrombosis.
  • Electron microscopy reveals critical details about thrombus architecture.
  • Further research can build on these findings to enhance understanding of thrombosis.

Frequently Asked Questions

What is the main advantage of this technique?
It allows a reproducible way to study many stages of thrombosis on a structural level.
What should beginners avoid during the procedure?
Over-handling the tissue in the surgical area to prevent bleeding and complications.
How is the injury site marked?
By loosely encircling the region between two surgical threads of 0.1 millimeter size.
What type of microscopy is used for analysis?
Electron microscopy is used to analyze the samples collected.
What components are studied in this protocol?
Components of platelets and the hemostatic system involved in thrombosis.
What is the focus of the study?
To determine how thrombus growth and architecture are affected at the cellular level.

본 프로토콜은 동맥 혈전증을 유도하기 위해 FeCl3 매개 손상을 사용하는 방법, 및 전자 현미경 분석을 위해 혈전증의 다양한 단계에서 동맥 손상 샘플을 수집하고 준비하는 방법을 설명합니다.

이 방법을 사용하면 혈소판과 지혈 시스템의 어떤 성분이 혈전증에 중요한지 결정할 수 있습니다. 또한 이 방법은 혈전 성장과 구조가 세포 수준에서 어떻게 영향을 받는지 결정할 수 있습니다. 이 기술의 가장 큰 장점은 구조적 수준에서 혈전증의 여러 단계를 재현 가능한 방법으로 연구 할 수 있다는 것입니다.

이 기술을 처음 접하는 사람은 수술 부위의 조직을 과도하게 취급하지 않아야 합니다. 이것은 출혈로 이어질 수 있으며 경동맥의 식별을 지나치게 복잡하게 만들 수 있습니다. 경동맥을 노출시킨 후 0.1mm 크기의 두 수술실 사이의 영역을 느슨하게 둘러싸서 손상 부위를 표시하기 시작합니다.

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