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将超分辨率显微镜引入核心设施的管理

Managing the Introduction of Super-Resolution Microscopy into a Core Facility.

作者信息

Kamykowski Jeffrey A, Storrie Brian

机构信息

Digital Microscopy Laboratory, Department of Physiology and Biophysics, Slot 505, University of Arkansas for Medical Sciences, 4301 West Markham St, Little Rock, AR, 72202, USA.

出版信息

Methods Mol Biol. 2017;1663:15-19. doi: 10.1007/978-1-4939-7265-4_2.

Abstract

Super resolution techniques place the resolution of fluorescence microscopy closer to the size of the underlying cell structure or molecular machine being studied. Structured illumination techniques will give users a set of tools that are close to their past experience and relatively simple and quick to learn. The present dyes can be used. Resolution approaching 100 nm XY can be achieved. In contrast, stochastic methods such as PALM/STORM typically require the choice of new dyes and a much greater learning curve to master the technology and calculations. However, a further fivefold resolution improvement is possible. Stimulated depletion techniques such as STED offer a third set of approaches that will again require the use of new dyes. All these approaches require substantial investment in new equipment and in user training. There is no free lunch in the search for better resolution.

摘要

超分辨率技术使荧光显微镜的分辨率更接近所研究的潜在细胞结构或分子机器的大小。结构照明技术将为用户提供一套与他们过去经验相近且相对简单易学的工具。可以使用现有的染料。能够实现接近100纳米XY的分辨率。相比之下,诸如PALM/STORM之类的随机方法通常需要选择新的染料,并且需要更大的学习曲线来掌握该技术和计算方法。然而,分辨率还有可能进一步提高五倍。诸如STED之类的受激发射损耗技术提供了另一套方法,这同样需要使用新的染料。所有这些方法都需要在新设备和用户培训方面进行大量投资。在追求更高分辨率的过程中没有免费的午餐。

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