Precise measurement of the resolution in light microscopy using Fourier transform

Arnold Vainrub

doi:10.1063/1.2912954

Precise measurement of the resolution in light microscopy using Fourier transform

Arnold Vainrub

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

4 Scopus citations

Abstract

The resolution power of light microscope has been accurately measured (±5%) by Fourier transform of various object images and further evaluation of the highest spatial frequency in Fourier spectrum. Any unknown shape plane object with a shape feature's size smaller than the resolution to be measured was shown to provide a reliable resolution test. This simple method gives a direct measurement of the resolution power as defined by Abbe [Archiv. F. Mikroskopische Anat. 9, 413 (1873)]. The results have been justified by comparison to a standard resolution measurement by using calibrated periodic line patterns. Notably, the approach is applicable in super-resolution light microscopy (transmission, reflection, and fluorescence), where calibrated resolution targets do not occur. It was conveniently implemented by using a compact disk as a test object and free IMAGEJ imaging software.

Original language	English (US)
Article number	046112
Journal	Review of Scientific Instruments
Volume	79
Issue number	4
DOIs	https://doi.org/10.1063/1.2912954
State	Published - 2008
Externally published	Yes

ASJC Scopus subject areas

Instrumentation

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10.1063/1.2912954

Cite this

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title = "Precise measurement of the resolution in light microscopy using Fourier transform",

abstract = "The resolution power of light microscope has been accurately measured (±5\%) by Fourier transform of various object images and further evaluation of the highest spatial frequency in Fourier spectrum. Any unknown shape plane object with a shape feature's size smaller than the resolution to be measured was shown to provide a reliable resolution test. This simple method gives a direct measurement of the resolution power as defined by Abbe [Archiv. F. Mikroskopische Anat. 9, 413 (1873)]. The results have been justified by comparison to a standard resolution measurement by using calibrated periodic line patterns. Notably, the approach is applicable in super-resolution light microscopy (transmission, reflection, and fluorescence), where calibrated resolution targets do not occur. It was conveniently implemented by using a compact disk as a test object and free IMAGEJ imaging software.",

author = "Arnold Vainrub",

note = "Funding Information: The support of the John E. Fetzer Memorial Fund (Grant No. 23-C) is acknowledged. I am grateful to Mats Gustafsson who via correspondence made clear to me the notions of resolution in imaging. I thank Vitaly Vodyanoy and Oleg Pustovyy for making the microphotographs of Richardson slide. A. Vainrub{\textquoteright} email is [email protected].",

year = "2008",

doi = "10.1063/1.2912954",

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journal = "Review of Scientific Instruments",

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AU - Vainrub, Arnold

N1 - Funding Information: The support of the John E. Fetzer Memorial Fund (Grant No. 23-C) is acknowledged. I am grateful to Mats Gustafsson who via correspondence made clear to me the notions of resolution in imaging. I thank Vitaly Vodyanoy and Oleg Pustovyy for making the microphotographs of Richardson slide. A. Vainrub’ email is [email protected].

PY - 2008

Y1 - 2008

N2 - The resolution power of light microscope has been accurately measured (±5%) by Fourier transform of various object images and further evaluation of the highest spatial frequency in Fourier spectrum. Any unknown shape plane object with a shape feature's size smaller than the resolution to be measured was shown to provide a reliable resolution test. This simple method gives a direct measurement of the resolution power as defined by Abbe [Archiv. F. Mikroskopische Anat. 9, 413 (1873)]. The results have been justified by comparison to a standard resolution measurement by using calibrated periodic line patterns. Notably, the approach is applicable in super-resolution light microscopy (transmission, reflection, and fluorescence), where calibrated resolution targets do not occur. It was conveniently implemented by using a compact disk as a test object and free IMAGEJ imaging software.

AB - The resolution power of light microscope has been accurately measured (±5%) by Fourier transform of various object images and further evaluation of the highest spatial frequency in Fourier spectrum. Any unknown shape plane object with a shape feature's size smaller than the resolution to be measured was shown to provide a reliable resolution test. This simple method gives a direct measurement of the resolution power as defined by Abbe [Archiv. F. Mikroskopische Anat. 9, 413 (1873)]. The results have been justified by comparison to a standard resolution measurement by using calibrated periodic line patterns. Notably, the approach is applicable in super-resolution light microscopy (transmission, reflection, and fluorescence), where calibrated resolution targets do not occur. It was conveniently implemented by using a compact disk as a test object and free IMAGEJ imaging software.

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Precise measurement of the resolution in light microscopy using Fourier transform

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