Sudan IV Staining: A Procedure for Staining Lipid-Laden Atherosclerotic Plaques in Harvested Mouse Aortic Arch
Published: April 30, 2023
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 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…
Divulgaciones
The authors have nothing to disclose.
Materials
| 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. |
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Sudan IV Staining: A Procedure for Staining Lipid-Laden Atherosclerotic Plaques in Harvested Mouse Aortic Arch. J. Vis. Exp. (Pending Publication), e21056, doi: (2023).