Effects of different dehydration methods on the preservation of aortic and renal glycocalyx structures in mice
Zhi Li,
Quanbin Zhang,
Yuan-yuan Sun,
Ning Wu
Affiliations
Zhi Li
CAS and Shandong Province Key Laboratory of Experimental Marine Biology, Center for Ocean Mega-Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China; Department of Drugs and Bioproducts, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China; University of Chinese Academy of Sciences, Beijing, China
Quanbin Zhang
CAS and Shandong Province Key Laboratory of Experimental Marine Biology, Center for Ocean Mega-Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China; Department of Drugs and Bioproducts, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China
Yuan-yuan Sun
CAS and Shandong Province Key Laboratory of Experimental Marine Biology, Center for Ocean Mega-Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China; Corresponding author.
Ning Wu
CAS and Shandong Province Key Laboratory of Experimental Marine Biology, Center for Ocean Mega-Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China; Department of Drugs and Bioproducts, Qingdao National Laboratory for Marine Science and Technology, Qingdao, China; Nantong Zhongke Marine Science and Technology Research and Development Center, Nantong, China; Corresponding author. Institute of Oceanology Chinese Academy of Sciences, China,
Glycocalyx is located outside the vascular endothelial cells playing an important role in vascular homeostasis. However, lacking efficient detection methods is one of the biggest obstacles to study the glycocalyx. In this study, three dehydration methods were used to compare the preservation of HUVEC, aorta and kidney glycocalyx by transmission electron microscope. The chemical pre-fixation was performed by lanthanum nitrate staining, and the mice aorta and renal glycocalyx were prepared by different dehydration methods such as ethanol gradient, acetone gradient and low temperature dehydration. HUVEC glycocalyx was prepared by acetone gradient and low temperature dehydration. Low temperature dehydration method preserves HUVEC and mice aortic glycocalyx completely, which had a certain thickness and presented a needle-like structure. But for mice kidney, the acetone gradient dehydration preparation method could better preserve the glycocalyx integrity than other two methods. In conclusion, low temperature dehydration method is suitable for HUVEC and aortic glycocalyx preservation, acetone gradient dehydration method is more suitable for kidney glycocalyx preservation.
