Sueiras Vivian M, Moy Vincent T, Ziebarth Noël M
Department of Biomedical Engineering, University of Miami College of Engineering, Coral Gables, FL.
Department of Physiology and Biophysics, University of Miami Miller School of Medicine, Miami, FL.
Mol Vis. 2015 Mar 15;21:316-23. eCollection 2015.
To image the ultrastructure of the anterior lens capsule at the nanoscale level using atomic force microscopy (AFM).
Experiments were performed on anterior lens capsules maintained in their in situ location surrounding the lens from six human cadavers (donor age range: 44-88 years), four cynomolgus monkeys (Macaca fascicularis age range: 4.83-8.92 years), and seven pigs (<6 months). Hydration of all samples was maintained using Dulbecco's Modified Eagle Medium (DMEM). Whole lenses were removed from the eye and placed anterior side up in agarose gel before gel hardening where only the posterior half of the lens was contained within the gel. After the gel hardened, the Petri dish was filled with DMEM until the point where the intact lens was fully submerged. AFM was used to image the anterior lens surface in contact mode. An integrated analysis program was used to calculate the interfibrillar spacing, fiber diameter, and surface roughness of the samples.
The AFM images depict a highly ordered fibrous structure at the surface of the lens capsule in all three species. The interfibrillar spacing for the porcine, cynomolgus monkey, and human lens capsules was 0.68±0.25, 1.80±0.39, and 1.08±0.25 μm, respectively. In the primate, interfibrillar spacing significantly decreased linearly as a function of age. The fiber diameters ranged from 50 to 950 nm. Comparison of the root mean square (RMS) and average deviation demonstrate that the surface of the porcine lens capsule is the smoothest, and that the human and cynomolgus monkey capsules are significantly rougher.
AFM was successful in providing high-resolution images of the nanostructure of the lens capsule samples. Species-dependent differences were observed in the overall structure and surface roughness.
使用原子力显微镜(AFM)在纳米尺度下对晶状体前囊膜的超微结构进行成像。
对取自6具人类尸体(供体年龄范围:44 - 88岁)、4只食蟹猴(猕猴年龄范围:4.83 - 8.92岁)和7头猪(<6个月)的晶状体前囊膜进行实验,这些前囊膜保持在晶状体原位周围。使用杜氏改良伊格尔培养基(DMEM)维持所有样本的水化状态。将完整晶状体从眼中取出,在琼脂糖凝胶硬化前正面朝上放置于凝胶中,此时凝胶仅包含晶状体的后半部分。凝胶硬化后,向培养皿中加入DMEM直至完整晶状体完全浸没。使用AFM以接触模式对晶状体前表面进行成像。使用综合分析程序计算样本的纤维间间距、纤维直径和表面粗糙度。
AFM图像显示所有三个物种的晶状体囊膜表面均为高度有序的纤维结构。猪、食蟹猴和人类晶状体囊膜的纤维间间距分别为0.68±0.25、1.80±0.39和1.08±0.25μm。在灵长类动物中,纤维间间距随年龄呈线性显著减小。纤维直径范围为50至950nm。均方根(RMS)和平均偏差的比较表明,猪晶状体囊膜表面最光滑,而人类和食蟹猴的囊膜明显更粗糙。
AFM成功提供了晶状体囊膜样本纳米结构的高分辨率图像。在整体结构和表面粗糙度方面观察到了物种依赖性差异。