期刊
METHODS AND APPLICATIONS IN FLUORESCENCE
卷 3, 期 1, 页码 -出版社
IOP PUBLISHING LTD
DOI: 10.1088/2050-6120/3/1/014003
关键词
localization microscopy; particle averaging; resolution improvement
资金
- NIH [R01R01GM100114]
- NSF [0954836]
- NewMexico Spatiotemporal Modeling Center [NIH P50GM0852673]
- Dutch Technology Foundation STW, Netherlands Organisation for Scientific Research (NWO)
- Ministry of Economic Affairs, Agriculture and Innovation
- Division Of Physics
- Direct For Mathematical & Physical Scien [0954836] Funding Source: National Science Foundation
Inspired by recent developments in localization microscopy that applied averaging of identical particles in 2D for increasing the resolution even further, we discuss considerations for alignment (registration) methods for particles in general and for 3D in particular. We detail that traditional techniques for particle registration from cryo electron microscopy based on cross-correlation are not suitable, as the underlying image formation process is fundamentally different. We argue that only localizations, i.e. a set of coordinates with associated uncertainties, are recorded and not a continuous intensity distribution. We present a method that owes to this fact and that is inspired by the field of statistical pattern recognition. In particular we suggest to use an adapted version of the Bhattacharyya distance as a merit function for registration. We evaluate the method in simulations and demonstrate it on 3D super-resolution data of Alexa 647 labelled to the Nup133 protein in the nuclear pore complex of Hela cells. From the simulations we find suggestions that for successful registration the localization uncertainty must be smaller than the distance between labeling sites on a particle. These suggestions are supported by theoretical considerations concerning the attainable resolution in localization microscopy and its scaling behavior as a function of labeling density and localization precision.
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