Atherosclerosis, a complex vascular disease characterized by plaque buildup within arteries, is a leading cause of cardiovascular diseases. Accurate collection and analysis of atherosclerotic lesions are crucial for advancing our understanding of disease progression and developing effective therapeutic strategies. In this JoVE collection, we detail a step-by-step protocol for the collection, preservation, and subsequent analysis of atherosclerotic specimens from animal models, providing valuable insights into lesion composition and molecular mechanisms.

This collection will encompass a wide range of methodological approaches, including but not limited to:

This collection encompasses a wide range of methodological approaches, including, but not limited to:

1.    Animal Model Preparation:

Selecting appropriate animal models (e.g., ApoE-/- mice) with established susceptibility to atherosclerosis.

2.    Tissue Harvesting:

Euthanizing animals using approved methods. Perfusing the cardiovascular system to remove blood contaminants and facilitate fixation. Excising target arteries (e.g., aorta, carotid) with minimal disruption. 

3.    Fixation and Preservation:

Optimizing fixation duration and the choice of fixative (e.g., paraformaldehyde) to maintain tissue morphology. Embedding specimens in paraffin or cryoprotectant for long-term preservation.

4.    Histological Processing:

Sectioning tissues to obtain thin slices suitable for staining and analysis. Performing routine histological staining (e.g., Hematoxylin & Eosin) to visualize general morphology.

5.    Advanced Staining Techniques:

Employing immunohistochemistry to detect specific cell types (macrophages, smooth muscle cells) and molecules (lipids, cytokines). Utilizing special stains like Oil Red O to highlight lipid-rich regions.

6.    Microscopic Analysis:

Capturing high-resolution images of stained sections. Quantifying key parameters such as plaque size, composition, and inflammatory cell infiltration.

7.    Data Interpretation:

Correlating histological findings with molecular data to gain insights into disease mechanisms.

Comparing results across experimental groups or treatment conditions.

 This JoVE video article introduces a state-of-the-art method for collecting and analyzing atherosclerotic tissues, offering researchers a robust tool to investigate disease progression and potential therapeutic targets. By combining advanced histological techniques with molecular analyses, this protocol contributes to a deeper understanding of atherosclerosis and its underlying biology.