论文标题
猪关节组织的三维成像向下到亚细胞水平
Three-dimensional imaging of porcine joint tissues down to the subcellular level
论文作者
论文摘要
关节组织由小梁和皮质骨以及钙化和透明软骨组成,这是由于局部X射线吸收值范围广泛的范围,这对在亚细胞水平上的基于硬X射线的可视化提出了挑战。钙化组织的密度需要相当高的光子能,这通常会导致软骨内的对比度不足,并阻碍了单个生物细胞的可视化。组织的脱钙化减少了总和X射线吸收值,并可以选择较低的光子能量。通过乙醇固定和石蜡组织嵌入可以实现进一步的对比度增强。在这项研究中,我们(i)搜索了一种适当的可视化方法,以研究激光骨质瘤产生的病变,(ii)在乙醇固定和随后使用基于实验室和同步子辐射的微辐射学的乙醇固定和随后的石蜡嵌入后,可视化了脱钙的猪关节。 Soleil的Anatomix光束线上的实验以1.3 um的像素大小进行离轴扫描模式进行。可以使关节所有层中的单个细胞可见,并显示出乙醇固定和石蜡嵌入的作用。
Joint tissues consist of trabecular and cortical bone as well as calcified and hyaline cartilage, which presents a challenge for hard X-ray-based visualization on the sub-cellular level due to the wide range of local X-ray absorption values. The density of the calcified tissues requires rather high photon energy, which often leads to insufficient contrast within the cartilage and impedes the visualization of individual biological cells. Decalcification of the tissues reduces the total and local X-ray absorption values and allows for selecting a lower photon energy. Further contrast enhancement can be achieved by ethanol fixation and paraffin tissue embedding. In this study, we (i) searched for an appropriate visualization method to investigate lesions generated by a laser osteotome and (ii) visualized a decalcified porcine joint after ethanol fixation and subsequent paraffin embedding using laboratory- and synchrotron radiation-based microtomography. The experiments at the ANATOMIX beamline of Synchrotron SOLEIL were performed in off-axis scan mode with a pixel size of 1.3 um. Individual cells in all layers of the joint could be made visible and the effect of ethanol fixation and paraffin embedding demonstrated.