We selected two rabbits among six KO founder rabbits for pathological analysis, which were 5♀ founder with LDLR/apoE double-KO and 7♂ founder with large fragment deletion. The aortic trees were isolated and opened out and, after fixing in formalin for 24 h, they were stained with Sudan IV (Wako Pure Chemical Industries Ltd., Osaka, Japan). For histological analysis, serial paraffin sections were stained with hematoxylin-eosin (HE) and immunohistochemically stained with monoclonal antibodies against either macrophages (clone: RAM11, Dako, Carpinteria, CA) or a-smooth muscle actin for smooth muscle cells (clone: HHF35, Dako, Carpinteria, CA).
Sudan 4
Manufactured by Fujifilm
Sourced in Japan
Sudan IV is a lab equipment product that functions as a biological stain. It is commonly used in histology and cytology procedures to detect and identify the presence of lipids in tissue samples.
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Lab products found in correlation
5 protocols using sudan 4
1
Pathological Analysis of Aortic Trees
2
Histological Analysis of Aortic Atherosclerosis
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Under anesthesia, PBS was perfused from left ventricle and then animals were killed and heart and aorta were dissected. Sudan IV (Wako: 192-04392) staining was conducted for aortic arch. Adventitial fat tissue was removed and aorta was dissected longitudinally. The image analysis software NIH Image (version 1.61; http://rsbweb.nih.gov/ij/ ) was used to calculate the ratio of the plaque lesion to the total aortic arch area.
Isolated thoracic aorta was fixed overnight with formalin at 4 °C. Tissue was routinely processed for paraffin embedding and 4 μm sections of thoracic aorta were cut and mounted on silanized slides and were stained by Hematoxylin Eosin (HE), alpha-smooth muscle actin (α-SMA) (NB300-978, RRID:AB_2273628) and CD68 (ab125212, RRID:AB_10975465, 1:500).
Isolated thoracic aorta was fixed overnight with formalin at 4 °C. Tissue was routinely processed for paraffin embedding and 4 μm sections of thoracic aorta were cut and mounted on silanized slides and were stained by Hematoxylin Eosin (HE), alpha-smooth muscle actin (α-SMA) (NB300-978, RRID:AB_2273628) and CD68 (ab125212, RRID:AB_10975465, 1:500).
3
Preparation of Artificially Soiled Fabric
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The materials used to prepare the artificially soiled fabric were cotton canequim (Laundry Science Association in Japan) , oleic acid (Fujifilm Wako Chemicals) , palmitic acid (Fujifilm Wako Chemicals) , and Sudan IV (Fujifilm Wako Chemicals) . Sodium hydroxide (Fujifilm Wako Chemicals, guaranteed grade) was used to adjust the pH of the cleaning solution, and sodium dodecyl sulfate (SDS, Fujifilm Wako Chemicals, for Biochemistry) was used as a surfactant.
4
Aortic Valve and Atherosclerosis Histology
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The heart and aorta were perfusion‐fixed with 4% paraformaldehyde. The aorta was excised from the root to the iliac artery and stained with Sudan IV (Catalog: 194‐07652Wako Pure Chemical Industries, Ltd.). The percent Sudan IV‐positive areas were measured using the image analyzer software, WinROOF (Mitani Corp), as previously reported, with some modifications.27 The transverse section of the aortic valve was stained with Oil Red O (Catalog: O0625‐25G; Sigma) and MOMA‐2 (Catalog: ab33451; Abcam, 1:1000 dilution), and the positively stained areas with the two stains were measured using the image analyzer software, Image J (NIH), and the positively stained areas were calculated, as previously reported, with some modifications.27
5
Quantifying Aortic Plaque and Inflammation
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Under anesthesia, PBS was perfused from left ventricle and then animals were killed and heart and aorta were dissected. Sudan IV (Wako: 192-04392) staining was conducted for aortic arch. Adventitial fat tissue was removed and aorta was dissected longitudinally. The image analysis software NIH Image (version 1.61; http://rsbweb.nih.gov/ij/ ) was used to calculate the ratio of the plaque lesion to the total aortic arch area.
Isolated heart was fixed overnight with formalin at 4 °C. Tissue was routinely processed for paraffin embedding and 4 μm sections of aortic root were cut and mounted on silanised slides and were stained by Masson trichrome, CD68 (ab125212, 1:500) and Mac-2 (CL8942AP, 1:2000). The positive areas were calculated as the ratio of the positively stained area to the total aortic valve area. Necrotic core areas, defined as areas primarily composed of lipids with cholesterol clefts, cellular debris, and foam cells, were determined from Masson trichrome stained sections.
Isolated heart was fixed overnight with formalin at 4 °C. Tissue was routinely processed for paraffin embedding and 4 μm sections of aortic root were cut and mounted on silanised slides and were stained by Masson trichrome, CD68 (ab125212, 1:500) and Mac-2 (CL8942AP, 1:2000). The positive areas were calculated as the ratio of the positively stained area to the total aortic valve area. Necrotic core areas, defined as areas primarily composed of lipids with cholesterol clefts, cellular debris, and foam cells, were determined from Masson trichrome stained sections.
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