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  Measuring image resolution in optical nanoscopy.

Nieuwenhuizen, R. P., Lidtke, K. A., Bates, M., Puig, D. L., Grünwald, D., Stallinga, S., et al. (2013). Measuring image resolution in optical nanoscopy. Nature Methods, 10(6), 557-562. doi:10.1038/NMETH.2448.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0013-F4AC-2 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-4838-3
Genre: Journal Article

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 Creators:
Nieuwenhuizen, R. P., Author
Lidtke, K. A., Author
Bates, M.1, Author              
Puig, D. L., Author
Grünwald, D., Author
Stallinga, S., Author
Rieger, B., Author
Affiliations:
1Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society, ou_578627              

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Free keywords: Nanoscale biophysics; Super-resolution microscopy; Fluorescence imaging; Cellular imaging
 Abstract: Resolution in optical nanoscopy (or super-resolution microscopy) depends on the localization uncertainty and density of single fluorescent labels and on the sample's spatial structure. Currently there is no integral, practical resolution measure that accounts for all factors. We introduce a measure based on Fourier ring correlation (FRC) that can be computed directly from an image. We demonstrate its validity and benefits on two-dimensional (2D) and 3D localization microscopy images of tubulin and actin filaments. Our FRC resolution method makes it possible to compare achieved resolutions in images taken with different nanoscopy methods, to optimize and rank different emitter localization and labeling strategies, to define a stopping criterion for data acquisition, to describe image anisotropy and heterogeneity, and even to estimate the average number of localizations per emitter. Our findings challenge the current focus on obtaining the best localization precision, showing instead how the best image resolution can be achieved as fast as possible.

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Language(s): eng - English
 Dates: 2013-04-282013-06
 Publication Status: Published in print
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1038/NMETH.2448
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Title: Nature Methods
Source Genre: Journal
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Pages: - Volume / Issue: 10 (6) Sequence Number: - Start / End Page: 557 - 562 Identifier: -