Lilledahl Magnus B, Stokke Bjørn T
Biophysics and Medical Technology, Department of Physics, The Norwegian University of Science and Technology Trondheim, Norway.
Front Microbiol. 2015 May 28;6:525. doi: 10.3389/fmicb.2015.00525. eCollection 2015.
Understanding of biology is underpinned by the ability to observe structures at various length scales. This is so in a historical context and is also valid today. Evolution of novel insight often emerges from technological advancement. Recent developments in imaging technologies that is relevant for characterization of extraceullar microbiological polysaccharides are summarized. Emphasis is on scanning probe and optical based techniques since these tools offers imaging capabilities under aqueous conditions more closely resembling the physiological state than other ultramicroscopy imaging techniques. Following the demonstration of the scanning probe microscopy principle, novel operation modes to increase data capture speed toward video rate, exploitation of several cantilever frequencies, and advancement of utilization of specimen mechanical properties as contrast, also including their mode of operation in liquid, have been developed on this platform. Combined with steps in advancing light microscopy with resolution beyond the far field diffraction limit, non-linear methods, and combinations of the various imaging modalities, the potential ultramicroscopy toolbox available for characterization of exopolysaccharides (EPS) are richer than ever. Examples of application of such ultramicroscopy strategies range from imaging of isolated microbial polysaccharides, structures being observed when they are involved in polyelectrolyte complexes, aspects of their enzymatic degradation, and cell surface localization of secreted polysaccharides. These, and other examples, illustrate that the advancement in imaging technologies relevant for EPS characterization supports characterization of structural aspects.
对生物学的理解建立在能够在各种长度尺度上观察结构的能力基础之上。这在历史背景下是如此,在今天也是有效的。新见解的产生往往源于技术进步。本文总结了与细胞外微生物多糖表征相关的成像技术的最新进展。重点是扫描探针和基于光学的技术,因为这些工具在水性条件下提供的成像能力比其他超微镜成像技术更接近生理状态。在演示了扫描探针显微镜原理之后,基于该平台开发了新的操作模式,以提高数据采集速度至视频速率,利用多种悬臂频率,以及利用样品的机械性能作为对比度,还包括它们在液体中的操作模式。结合提高光学显微镜分辨率超越远场衍射极限的步骤、非线性方法以及各种成像方式的组合,可用于表征胞外多糖(EPS)的潜在超微镜工具箱比以往任何时候都更加丰富。这种超微镜策略的应用实例包括对分离的微生物多糖的成像、它们参与聚电解质复合物时观察到的结构、它们的酶促降解方面以及分泌多糖的细胞表面定位。这些以及其他例子表明,与EPS表征相关的成像技术的进步支持了对结构方面的表征。