Sudan IV Staining: A Procedure for Staining Lipid-Laden Atherosclerotic Plaques in Harvested Mouse Aortic Arch

Abstract

Source: Centa, M., et al. Quantification of Atherosclerosis in Mice. J. Vis. Exp. (2019)

This video demonstrates a procedure using Sudan IV stain for staining the fatty deposits in the harvested mouse aortic arch to visualize the atherosclerotic plaques and determine the extent of atherosclerosis.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board

1. En Face Analysis of Aortic Arch and Brachiocephalic Artery

1. Prepare pinning beds for en face analysis of aortic arches. Fold a segment of paraffin wax film eight times to make a flat 25 mm x 25 mm surface. Wrap it with black electric insulation tape to make a dark background for the aorta. Place a label on the backside of the pinning bed and use a lead pencil to write the mouse identification number (normal pen ink will disappear in the staining process).
2. Tra....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Black electrical insulation tape (50 mm wide)	Any specialized retailer		To create pinning beds for aortic arches.
Dissecting scissors (10 cm, straight)	World Precision Instruments	14393	For general dissection of organs.
Dumont forceps #5 (11 cm, straight)	World Precision Instruments	500341	For microdissection of aorta.
Ethanol 70% (v/v)	VWR Chemicals	83801.29	Highly flammable liquid and vapour, store in a well-ventilated place, and keep cool.
Ethanol absolute ≥99.8%	VWR Chemicals	20821.31	Highly flammable liquid and vapour, store in a well-ventilated place, and keep cool.
Formaldehyde 4% stabilised, buffered (pH 7.0)	VWR Chemicals	9713.1	Harmful by inhalation, in contact with skin and if swallowed.
Isopropanol	Merck	1096341011	Flammable liquid, causes serious eye irritation, and may cause drowsiness or dizziness.
Micro Castroviejo needle holder (9 cm, straight)	World Precision Instruments	503376	For pinning of aortic arches.
Minutien insect pins, 0.10 mm	Fine Science Tools	26002-10	For pinning of aortic arches.
Parafilm M	Bemis	PM992	Paraffin wax film used to create pinning beds for aortic arches.
Phosphate buffered saline (PBS)	-	-	Sterile and RNase-free solution is required for perfusion of mice
Stereomicroscope	Leica Microsystems	MZ6	For dissection and en face documentation
Sudan IV	Sigma-Aldrich	S4261	Not classified as a hazardous substance or mixture.
Vannas scissors (8 cm, straight)	World Precision Instruments	503378	For microdissection of aorta.