Center for Nanophase Materials Sciences, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA; E-Mails:
Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA; E-Mails:
Biosensors (Basel). 2014 Mar 14;4(1):63-75. doi: 10.3390/bios4010063. eCollection 2014 Mar.
The attachment and arrangement of microbes onto a substrate is influenced by both the biochemical and physical surface properties. In this report, we develop lectin-functionalized substrates containing patterned, three-dimensional polymeric structures of varied shapes and densities and use these to investigate the effects of topology and spatial confinement on lectin-mediated microbe immobilization. Films of poly(glycidyl methacrylate)-block-4,4-dimethyl-2-vinylazlactone (PGMA-b-PVDMA) were patterned on silicon surfaces into line arrays or square grid patterns with 5 μm wide features and varied pitch. The patterned films had three-dimensional geometries with 900 nm film thickness. After surface functionalization with wheat germ agglutinin, the size of Pseudomonas fluorescens aggregates immobilized was dependent on the pattern dimensions. Films patterned as parallel lines or square grids with a pitch of 10 μm or less led to the immobilization of individual microbes with minimal formation of aggregates. Both geometries allowed for incremental increases in aggregate size distribution with each increase in pitch. These engineered surfaces combine spatial confinement with affinity-based capture to control the extent of microbe adhesion and aggregation, and can also be used as a platform to investigate intercellular interactions and biofilm formation in microbial populations of controlled sizes.
微生物在基质上的附着和排列受到生化和物理表面特性的影响。在本报告中,我们开发了带有图案的、具有不同形状和密度的三维聚合物结构的凝集素功能化基底,并使用这些基底来研究拓扑和空间限制对凝集素介导的微生物固定化的影响。聚(甲基丙烯酸缩水甘油酯)-嵌段-4,4-二甲基-2-乙烯基恶唑啉(PGMA-b-PVDMA)薄膜在硅表面上形成线阵列或具有 5 μm 宽特征和不同节距的正方形网格图案。图案化的薄膜具有 900nm 厚的三维几何形状。经过麦胚凝集素表面功能化后,固定化荧光假单胞菌聚集体的大小取决于图案尺寸。具有 10μm 或更小节距的平行线条或正方形网格图案导致单个微生物的固定化,聚集体的形成最小。这两种几何形状都允许通过每次增加节距来逐渐增加聚集体的大小分布。这些工程表面将空间限制与基于亲和力的捕获结合起来,以控制微生物附着和聚集的程度,还可以用作研究具有受控大小的微生物群体中的细胞间相互作用和生物膜形成的平台。