Evaluation resolution in live cell structured illumination microscopy

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68407700%3A21230%2F19%3A00332792" target="_blank" >RIV/68407700:21230/19:00332792 - isvavai.cz</a>

Výsledek na webu

<a href="https://doi.org/10.1117/12.2527885" target="_blank" >https://doi.org/10.1117/12.2527885</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1117/12.2527885" target="_blank" >10.1117/12.2527885</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Evaluation resolution in live cell structured illumination microscopy

Popis výsledku v původním jazyce

In the last decade, several different structured illumination microscopy (SIM) approaches have been developed. Precise determination of the effective spatial resolution in a live cell SIM reconstructed image is essential for reliable interpretation of reconstruction results. Theoretical resolution improvement can be calculated for every SIM method. In practice, the final spatial resolution of the cell structures in the reconstructed image is limited by many different factors. Therefore, assessing the resolution directly from the single image is an inherent part of the live cell imaging. There are several commonly used resolution measurement techniques based on image analysis. These techniques include full-width at half maximum (FWHM) criterion, or Fourier ring correlation (FRC). FWHM measurement requires fluorescence beads or sharp edge/line in the observed image to determine the point spread function (PSF). FRC method requires two stochastically independent images of the same observed sample. Based on our experimental findings, the FRC method does not seem to be well suited for measuring the resolution of SIM live cell video sequences. Here we show a method based on the Fourier transform analysis using power spectral density (PSD). In order to estimate the cut-off frequency from a noisy signal, we use PSD estimation based on Welch’s method. This method is widely used in non-parametric power spectra analysis. Since the PSD-based metric can be computed from a single SIM image (one video frame), without any prior knowledge of the acquiring system, it can become a fundamental tool for imaging in live cell biology.

Název v anglickém jazyce

Evaluation resolution in live cell structured illumination microscopy

Popis výsledku anglicky

In the last decade, several different structured illumination microscopy (SIM) approaches have been developed. Precise determination of the effective spatial resolution in a live cell SIM reconstructed image is essential for reliable interpretation of reconstruction results. Theoretical resolution improvement can be calculated for every SIM method. In practice, the final spatial resolution of the cell structures in the reconstructed image is limited by many different factors. Therefore, assessing the resolution directly from the single image is an inherent part of the live cell imaging. There are several commonly used resolution measurement techniques based on image analysis. These techniques include full-width at half maximum (FWHM) criterion, or Fourier ring correlation (FRC). FWHM measurement requires fluorescence beads or sharp edge/line in the observed image to determine the point spread function (PSF). FRC method requires two stochastically independent images of the same observed sample. Based on our experimental findings, the FRC method does not seem to be well suited for measuring the resolution of SIM live cell video sequences. Here we show a method based on the Fourier transform analysis using power spectral density (PSD). In order to estimate the cut-off frequency from a noisy signal, we use PSD estimation based on Welch’s method. This method is widely used in non-parametric power spectra analysis. Since the PSD-based metric can be computed from a single SIM image (one video frame), without any prior knowledge of the acquiring system, it can become a fundamental tool for imaging in live cell biology.

Druh

D - Stať ve sborníku

CEP obor

—

OECD FORD obor

20201 - Electrical and electronic engineering

Projekt

<a href="/cs/project/GA17-05840S" target="_blank" >GA17-05840S: Multikriteriální optimalizace modelů prostorově variantních zobrazovacích systémů</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název statě ve sborníku

Proceedings Volume 11137 - Applications of Digital Image Processing XLII

ISBN

978-1-5106-2967-7

ISSN

0277-786X

e-ISSN

1996-756X

Počet stran výsledku

9

Strana od-do

—

Název nakladatele

SPIE

Místo vydání

Bellingham

Místo konání akce

San Diego, California

Datum konání akce

11. 8. 2019

Typ akce podle státní příslušnosti

WRD - Celosvětová akce

Kód UT WoS článku

000511298300007