Characterisation of atherosclerotic lesions with scanning electron microscopy (SEM) of wet tissue.
Atherosclerotic cardiovascular disease (CVD) is the universal leading cause of mortality and a major cause of morbidity. Additionally, the global epidemic of diabetes is associated with considerable cardiovascular mortality risk due to accelerated premature atherosclerosis. Development of effective therapies for atherosclerosis is dependent upon improved tools to assess atherosclerotic lesion progression in animal models. We present a novel technique that utilises scanning electron microscopy (SEM) for imaging wet biological specimens, thus enabling rapid and high-resolution imaging of atherosclerotic lesions. This wet SEM technique was used in an apoE-deficient mice model for morphological characterisation of early and advanced atherosclerotic lesions. Further demonstration of lipid-rich atherosclerotic lesions was carried out with osmium tetroxide staining for cholesterol. Gold immunolabelling of specific epitopes was applied in identification of the cellular and molecular components within the atherosclerotic lesions, namely foam cells, smooth muscle cells and collagen. The wet SEM technique demonstrates an accurate and detailed structural evaluation of the pathological process of atherosclerosis. Understanding the mechanisms that precipitate the atherosclerotic process, utilising this novel technique, may assist in the development of innovative therapeutic interventions for CVD management and prevention in the general population and in those with diabetes.
Kamari
Yehuda
Y
Tel-Hashomer and Sackler Faculty of Medicine, Tel Aviv University, Israel.
Cohen
Hofit
H
Shaish
Aviv
A
Bitzur
Rafael
R
Afek
Arnon
A
Shen
Shlomzion
S
Vainshtein
Anya
A
Harats
Dror
D
D000818:Animals; D001011:Aorta / Q000378:metabolism / Q000473:pathology; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D050197:Atherosclerosis / Q000378:metabolism* / Q000473:pathology*; D002784:Cholesterol / Q000378:metabolism*; D003094:Collagen / Q000378:metabolism; D004195:Disease Models, Animal; D000939:Epitopes; D005260:Female; D005487:Foam Cells / Q000166:cytology / Q000378:metabolism; D007150:Immunohistochemistry; D008264:Macrophages / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008855:Microscopy, Electron, Scanning / Q000379:methods*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism
D001057:Apolipoproteins E; D000939:Epitopes; D003094:Collagen; D002784:Cholesterol