Randall Division of Cell & Molecular Biophysics, King's College London, London, UK.
Int J Biochem Cell Biol. 2013 Aug;45(8):1669-78. doi: 10.1016/j.biocel.2013.05.010. Epub 2013 May 18.
Recently developed super-resolution techniques in optical microscopy have pushed the length scale at which cellular structure can be observed down to tens of nanometres. A wide array of methods have been described that fall under the umbrella term of super-resolution microscopy and each of these methods has different requirements for acquisition speed, experimental complexity, fluorophore requirements and post-processing of data. For example, experimental complexity can be decreased by using a standard widefield microscope for acquisition, but this requires substantial processing of the data to extract the super-resolution information. These powerful techniques are bringing new insights into the nanoscale structure of sub-cellular assemblies such as podosomes, which are an ideal system to observe with super-resolution microscopy as the structures are relatively thin and they form and dissociate over a period of several minutes. Here we discuss the major classes of super-resolution microscopy techniques, and demonstrate their relative performance by imaging podosomes.
近年来，光学显微镜中的超分辨率技术已经将可观察到的细胞结构的长度尺度推至几十纳米。已经描述了广泛的方法，这些方法都属于超分辨率显微镜这一统称，每种方法对采集速度、实验复杂性、荧光染料要求和数据后处理都有不同的要求。例如，通过使用标准的宽场显微镜进行采集，可以降低实验的复杂性，但这需要对数据进行大量处理才能提取超分辨率信息。这些强大的技术正在为细胞亚结构的纳米尺度结构带来新的认识，例如足突，这是一个用超分辨率显微镜观察的理想系统，因为这些结构相对较薄，并且在几分钟的时间内形成和分解。在这里，我们讨论了超分辨率显微镜技术的主要类别，并通过对足突进行成像来展示它们的相对性能。
