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SEM observation

ZW Zongyi Wu
BW Bingzhang Wang
JT Jiahao Tang
BB Bingli Bai
SW Sheji Weng
ZX Zhongjie Xie
ZS Zijian Shen
DY Deyi Yan
LC Liang Chen
JZ Jingdong Zhang
LY Lei Yang
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Before SEM observation, frozen section was performed to cut the surface produced by band saw in order to obtain an observation surface that was less of mechanical damage. After washed by phosphate-buffered saline (PBS), the samples were dehydrated in a series of graded concentration of ethanol from 70 to 100% and soaked in tert-butyl alcohol. Lyophilization was performed with a vacuum freeze drier (Christ Delta 2-24 LSC, Osterode, Germany) subsequently. Then the samples were Au sputter-coated (Eiko VX-10, Tokyo, Japan) and examined under SEM (S-3000N, HITACHI, Japan). In order to obtain more data and decrease error, five observation points were set (Fig. ​(Fig.1c).1c). The diameter of collagen fibers and gap between collagen fibers were measured by ImageJ (version 1.8.0). Then, the mean values were calculated and compared in histograms.

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