Stenger D A, Hickman J J, Bateman K E, Ravenscroft M S, Ma W, Pancrazio J J, Shaffer K, Schaffner A E, Cribbs D H, Cotman C W
Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, Washington, DC 20375, USA.
J Neurosci Methods. 1998 Aug 1;82(2):167-73. doi: 10.1016/s0165-0270(98)00047-8.
High resolution substrates, created using patterned self-assembled monolayers, are shown to direct axonal and dendritic process extension at the level of a single hippocampal neuron. Axons and dendrites were identified using morphological characteristics and immunocytochemical markers. Patterns were formed on glass coverslips from a co-planar monolayer of cell adhesive aminosilanes and non-adhesive fluorinated silanes. On patterned surfaces, the percentage of the total number of cells attached to the 0.71 mm2 substrate field with compliance to the 25-micron diameter 'somal adhesion site' reached 41 +/- 7% (mean +/- S.D., 428 cells counted). A total of 76 +/- 11% of cells that adhered to a somal attachment site developed a lone process > or = 100 microns oriented in the direction of the continuous aminosilane pathway which was shown to express axonal markers. Cells on either the fluorinated silane, which is non-permissive for neurite outgrowth, or localized on an aminosilane region only 5 microns wide failed to extend major processes. This approach is amenable to a variety of industry standard fabrication techniques and may be used to study the role of fine scale spatial cues in neuronal development and synapse formation.
使用图案化自组装单分子层创建的高分辨率底物，被证明能在单个海马神经元水平上引导轴突和树突的生长。通过形态学特征和免疫细胞化学标记物来识别轴突和树突。图案是由细胞粘附性氨基硅烷和非粘附性氟化硅烷的共平面单分子层在玻璃盖玻片上形成的。在图案化表面上，附着在0.71平方毫米底物区域且符合25微米直径“体细胞粘附位点”的细胞总数百分比达到41±7%（平均值±标准差，共计数428个细胞）。总共76±11%附着在体细胞附着位点的细胞形成了一个长度≥100微米的单一突起，其方向与连续氨基硅烷途径一致，该途径显示表达轴突标记物。在氟化硅烷（不允许神经突生长）上的细胞，或仅位于5微米宽的氨基硅烷区域内的细胞，均未能延伸出主要突起。这种方法适用于各种行业标准制造技术，可用于研究精细尺度空间线索在神经元发育和突触形成中的作用。
