Fixed Pattern Noise in Localization Microscopy

Patrick Fox-Roberts; Tianqi Wen; Klaus Suhling; Susan Cox

doi:10.1002/cphc.201300756

Fixed Pattern Noise in Localization Microscopy

Patrick Fox-Roberts, Tianqi Wen, Klaus Suhling, Susan Cox^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

4 Citations (Scopus)

Abstract

Localization microscopy vastly improves the resolution achieved by fluorescence microscopy by fitting the positions of individual fluorophores. We examine the reconstructions produced by different fitting algorithms for instances of fixed pattern noisesystematic tendencies to alter estimated emitter positions according to their subpixel location in a way that does not reflect the ground truth structure. We show that while not readily visible at standard empirical signal strengths, fixed pattern noise can occur when performing sub-pixel fitting, and that its degree varies according to the algorithm used and the relative size of the pixels compared to the point spread function. For pixel sizes in the range 80-170 nm, this results in variations in accuracy of the order of 2-4 nmcomparatively small for many applications, but non-negligible in scenarios where very high accuracy is sought.

Original language	English
Pages (from-to)	677-686
Number of pages	10
Journal	ChemPhysChem
Volume	15
Issue number	4
DOIs	https://doi.org/10.1002/cphc.201300756
Publication status	Published - 17 Mar 2014

Keywords

algorithms
fixed pattern noise
fluorescence
localization microscopy
super-resolution
SINGLE-MOLECULE LOCALIZATION
SUPERRESOLUTION MICROSCOPY
FLUORESCENT-PROBES
HIGH-RESOLUTION
ACCURACY
NANOSCOPY
TRACKING

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10.1002/cphc.201300756

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title = "Fixed Pattern Noise in Localization Microscopy",

abstract = "Localization microscopy vastly improves the resolution achieved by fluorescence microscopy by fitting the positions of individual fluorophores. We examine the reconstructions produced by different fitting algorithms for instances of fixed pattern noisesystematic tendencies to alter estimated emitter positions according to their subpixel location in a way that does not reflect the ground truth structure. We show that while not readily visible at standard empirical signal strengths, fixed pattern noise can occur when performing sub-pixel fitting, and that its degree varies according to the algorithm used and the relative size of the pixels compared to the point spread function. For pixel sizes in the range 80-170 nm, this results in variations in accuracy of the order of 2-4 nmcomparatively small for many applications, but non-negligible in scenarios where very high accuracy is sought.",

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author = "Patrick Fox-Roberts and Tianqi Wen and Klaus Suhling and Susan Cox",

year = "2014",

month = mar,

day = "17",

doi = "10.1002/cphc.201300756",

language = "English",

volume = "15",

pages = "677--686",

journal = "ChemPhysChem",

issn = "1439-4235",

publisher = "Wiley-Blackwell",

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T1 - Fixed Pattern Noise in Localization Microscopy

AU - Fox-Roberts, Patrick

AU - Wen, Tianqi

AU - Suhling, Klaus

AU - Cox, Susan

PY - 2014/3/17

Y1 - 2014/3/17

N2 - Localization microscopy vastly improves the resolution achieved by fluorescence microscopy by fitting the positions of individual fluorophores. We examine the reconstructions produced by different fitting algorithms for instances of fixed pattern noisesystematic tendencies to alter estimated emitter positions according to their subpixel location in a way that does not reflect the ground truth structure. We show that while not readily visible at standard empirical signal strengths, fixed pattern noise can occur when performing sub-pixel fitting, and that its degree varies according to the algorithm used and the relative size of the pixels compared to the point spread function. For pixel sizes in the range 80-170 nm, this results in variations in accuracy of the order of 2-4 nmcomparatively small for many applications, but non-negligible in scenarios where very high accuracy is sought.

AB - Localization microscopy vastly improves the resolution achieved by fluorescence microscopy by fitting the positions of individual fluorophores. We examine the reconstructions produced by different fitting algorithms for instances of fixed pattern noisesystematic tendencies to alter estimated emitter positions according to their subpixel location in a way that does not reflect the ground truth structure. We show that while not readily visible at standard empirical signal strengths, fixed pattern noise can occur when performing sub-pixel fitting, and that its degree varies according to the algorithm used and the relative size of the pixels compared to the point spread function. For pixel sizes in the range 80-170 nm, this results in variations in accuracy of the order of 2-4 nmcomparatively small for many applications, but non-negligible in scenarios where very high accuracy is sought.

KW - algorithms

KW - fixed pattern noise

KW - fluorescence

KW - localization microscopy

KW - super-resolution

KW - SINGLE-MOLECULE LOCALIZATION

KW - SUPERRESOLUTION MICROSCOPY

KW - FLUORESCENT-PROBES

KW - HIGH-RESOLUTION

KW - ACCURACY

KW - NANOSCOPY

KW - TRACKING

U2 - 10.1002/cphc.201300756

DO - 10.1002/cphc.201300756

M3 - Article

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VL - 15

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EP - 686

JO - ChemPhysChem

JF - ChemPhysChem

IS - 4

ER -

Fixed Pattern Noise in Localization Microscopy

Abstract

Keywords

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