Centro Brasileiro de Pesquisas Físicas, Xavier Sigaud, 150, CEP 22290-180 Rio de Janeiro, Brazil; Universidade Federal do Rio de Janeiro, Centro de Ciências da Saúde, Instituto de Ciências Biomédicas, Laboratório de Biomineralização, CEP 21941-590 Rio de Janeiro, Brazil.
Instituto Nacional de Metrologia, Qualidade e Tecnologia, Divisão de Metrologia de Materiais, Duque de Caxias, CEP 25250-020 Rio de Janeiro, Brazil.
Acta Biomater. 2014 Sep;10(9):3875-84. doi: 10.1016/j.actbio.2014.01.023. Epub 2014 Jan 30.
We investigated the ultrastructure and crystallographic orientation of spicules from the calcareous sponge Paraleucilla magna (subclass Calcaronea) by transmission and scanning electron microscopy using two different methods of sample preparation: ultramicrotomy and focused ion beam (FIB). It was found that the unpaired actine from the spicules was oriented in the [211] zone axis. The plane that contains the unpaired actine and divides symmetrically the paired actines is the (-120). This plane is a mirror plane of the hexagonal lattice system. All the spicule types analyzed presented the same crystallographic orientation. Electron nanodiffraction maps from 4μm×4μm regions prepared by FIB showed disorientation of <2° between diffraction patterns obtained from neighbor regions, indicating the presence of a unique, highly aligned calcite crystalline phase. Among the eight FIB sections obtained, four presented high pore density. In one section perpendicular to the actine axis pores were observed only in the center of the spicule aligned in a circular pattern and surrounded by a faint circular contour with a larger radius. The presence of amorphous carbon representative of organic molecules detected by electron energy loss spectroscopy was correlated neither with porosity nor with specific lattice planes.
我们通过透射电子显微镜和扫描电子显微镜,使用两种不同的样品制备方法——超薄切片和聚焦离子束(FIB),研究了钙质海绵 Paraleucilla magna(亚纲 Calcaronea)的骨针的超微结构和结晶取向。结果发现,未配对的肌动蛋白在骨针中沿[211]晶带轴取向。包含未配对的肌动蛋白并将配对的肌动蛋白对称分割的平面是(-120)。该平面是六方晶格系统的镜面。分析的所有骨针类型均呈现相同的结晶取向。通过 FIB 制备的 4μm×4μm 区域的电子纳米衍射图谱显示,来自相邻区域的衍射图谱之间的取向偏差小于 2°,表明存在独特的、高度有序的方解石结晶相。在所获得的八个 FIB 切片中,有四个呈现出高孔隙密度。在一个垂直于肌动蛋白轴的切片中,仅在沿圆形图案排列并被具有更大半径的微弱圆形轮廓包围的骨针中心观察到孔。通过电子能量损失光谱检测到的代表有机分子的无定形碳既与孔隙率无关,也与特定晶格平面无关。