Novel Video-FRAP method for the determination of anisotropic diffusion: Principles and optimization

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Fluorescence recovery after photobleaching (FRAP) is an established technique for determining solute diffusivity in biological tissues. Recently, we proposed a novel fluorescence photobleaching method, based on two independent analyses of the video-FRAP image series: the fast Fourier transform (FFT) and the Karhunen-Loève transform (KLT). In this study, the principles of this method are illustrated, and its optimization is discussed. Computer simulated FRAP experiments were used for the optimization of the choice of frequencies used in FFT analysis. The sensitivity of the technique to image noise contamination was evaluated. Additionally, the method was experimentally validated by characterizing fluorescein diffusivity in human annulus fibrosus. The results were compared to diffusivity data obtained by other methods. This study provides new insights into the optimization of data analysis of video-FRAP experiments. Furthermore, the experimental results verified the applicability of the method for the determination of solute diffusivity in biological media.

Original languageEnglish
Title of host publicationIFMBE Proceedings
Pages9-10
Number of pages2
Volume24
DOIs
StatePublished - Nov 6 2009
Event25th Southern Biomedical Engineering Conference 2009 - Miami, FL, United States
Duration: May 15 2009May 17 2009

Other

Other25th Southern Biomedical Engineering Conference 2009
CountryUnited States
CityMiami, FL
Period5/15/095/17/09

Fingerprint

Photobleaching
Fluorescence
Recovery
Fast Fourier transforms
Fluorescein
Contamination
Experiments
Tissue

Keywords

  • Annulus Fibrosus (AF)
  • Fast Fourier Transform (FFT)
  • Gaussian noise

ASJC Scopus subject areas

  • Biomedical Engineering
  • Bioengineering

Cite this

Novel Video-FRAP method for the determination of anisotropic diffusion : Principles and optimization. / Travascio, Francesco; Gu, Weiyong.

IFMBE Proceedings. Vol. 24 2009. p. 9-10.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Travascio, F & Gu, W 2009, Novel Video-FRAP method for the determination of anisotropic diffusion: Principles and optimization. in IFMBE Proceedings. vol. 24, pp. 9-10, 25th Southern Biomedical Engineering Conference 2009, Miami, FL, United States, 5/15/09. https://doi.org/10.1007/978-3-642-01697-4_5
@inproceedings{4a5c5a0509fa43f6babe8387988217d6,
title = "Novel Video-FRAP method for the determination of anisotropic diffusion: Principles and optimization",
abstract = "Fluorescence recovery after photobleaching (FRAP) is an established technique for determining solute diffusivity in biological tissues. Recently, we proposed a novel fluorescence photobleaching method, based on two independent analyses of the video-FRAP image series: the fast Fourier transform (FFT) and the Karhunen-Lo{\`e}ve transform (KLT). In this study, the principles of this method are illustrated, and its optimization is discussed. Computer simulated FRAP experiments were used for the optimization of the choice of frequencies used in FFT analysis. The sensitivity of the technique to image noise contamination was evaluated. Additionally, the method was experimentally validated by characterizing fluorescein diffusivity in human annulus fibrosus. The results were compared to diffusivity data obtained by other methods. This study provides new insights into the optimization of data analysis of video-FRAP experiments. Furthermore, the experimental results verified the applicability of the method for the determination of solute diffusivity in biological media.",
keywords = "Annulus Fibrosus (AF), Fast Fourier Transform (FFT), Gaussian noise",
author = "Francesco Travascio and Weiyong Gu",
year = "2009",
month = "11",
day = "6",
doi = "10.1007/978-3-642-01697-4_5",
language = "English",
isbn = "9783642016967",
volume = "24",
pages = "9--10",
booktitle = "IFMBE Proceedings",

}

TY - GEN

T1 - Novel Video-FRAP method for the determination of anisotropic diffusion

T2 - Principles and optimization

AU - Travascio, Francesco

AU - Gu, Weiyong

PY - 2009/11/6

Y1 - 2009/11/6

N2 - Fluorescence recovery after photobleaching (FRAP) is an established technique for determining solute diffusivity in biological tissues. Recently, we proposed a novel fluorescence photobleaching method, based on two independent analyses of the video-FRAP image series: the fast Fourier transform (FFT) and the Karhunen-Loève transform (KLT). In this study, the principles of this method are illustrated, and its optimization is discussed. Computer simulated FRAP experiments were used for the optimization of the choice of frequencies used in FFT analysis. The sensitivity of the technique to image noise contamination was evaluated. Additionally, the method was experimentally validated by characterizing fluorescein diffusivity in human annulus fibrosus. The results were compared to diffusivity data obtained by other methods. This study provides new insights into the optimization of data analysis of video-FRAP experiments. Furthermore, the experimental results verified the applicability of the method for the determination of solute diffusivity in biological media.

AB - Fluorescence recovery after photobleaching (FRAP) is an established technique for determining solute diffusivity in biological tissues. Recently, we proposed a novel fluorescence photobleaching method, based on two independent analyses of the video-FRAP image series: the fast Fourier transform (FFT) and the Karhunen-Loève transform (KLT). In this study, the principles of this method are illustrated, and its optimization is discussed. Computer simulated FRAP experiments were used for the optimization of the choice of frequencies used in FFT analysis. The sensitivity of the technique to image noise contamination was evaluated. Additionally, the method was experimentally validated by characterizing fluorescein diffusivity in human annulus fibrosus. The results were compared to diffusivity data obtained by other methods. This study provides new insights into the optimization of data analysis of video-FRAP experiments. Furthermore, the experimental results verified the applicability of the method for the determination of solute diffusivity in biological media.

KW - Annulus Fibrosus (AF)

KW - Fast Fourier Transform (FFT)

KW - Gaussian noise

UR - http://www.scopus.com/inward/record.url?scp=70350614578&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=70350614578&partnerID=8YFLogxK

U2 - 10.1007/978-3-642-01697-4_5

DO - 10.1007/978-3-642-01697-4_5

M3 - Conference contribution

AN - SCOPUS:70350614578

SN - 9783642016967

VL - 24

SP - 9

EP - 10

BT - IFMBE Proceedings

ER -