Complex homogeneous and heterogeneous fluorescence anisotropy decays: Enhancing analysis accuracy

Željko ̌ Bajzer, M. C C Moncrieffe, Ivo Penzar, Franklyn G. Prendergast

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

In biological macromolecules, fluorophores often exhibit multiple depolarizing motions that require multiple lifetimes and rotational relaxation times to define fluorescence intensity and anisotropy decays. The related analysis of time-correlated single-photon counting data becomes uncertain due to the multitude of decay parameters and numerical sensitivity to deconvolution of the instrument response function (IRF) via discretization of integrals. By using simulations we show that improved discretizations based on quadratic and cubic local approximations of the IRF yield more accurate estimation of short rotational relaxation times and lifetimes than the commonly used Grinvald-Steinberg discretization, which in turn appears more reliable than two discretizations based on linear local approximations of the IRF. In addition, our simulation suggests that cubic approximation is the most advantageous in discriminating complex heterogeneous and homogeneous anisotropy decay. We show that among three different information criteria, the Akaike information criterion is best suited for detection of heterogeneity in rotational relaxation times. It is capable of detecting heterogeneity even when anisotropy decay appears homogeneous within statistical errors of estimation.

Original languageEnglish (US)
Pages (from-to)1765-1775
Number of pages11
JournalBiophysical Journal
Volume81
Issue number3
StatePublished - 2001

Fingerprint

Fluorescence Polarization
Anisotropy
Photons

ASJC Scopus subject areas

  • Biophysics

Cite this

Bajzer, Ž. ., Moncrieffe, M. C. C., Penzar, I., & Prendergast, F. G. (2001). Complex homogeneous and heterogeneous fluorescence anisotropy decays: Enhancing analysis accuracy. Biophysical Journal, 81(3), 1765-1775.

Complex homogeneous and heterogeneous fluorescence anisotropy decays : Enhancing analysis accuracy. / Bajzer, Željko ̌; Moncrieffe, M. C C; Penzar, Ivo; Prendergast, Franklyn G.

In: Biophysical Journal, Vol. 81, No. 3, 2001, p. 1765-1775.

Research output: Contribution to journalArticle

Bajzer, Ž, Moncrieffe, MCC, Penzar, I & Prendergast, FG 2001, 'Complex homogeneous and heterogeneous fluorescence anisotropy decays: Enhancing analysis accuracy', Biophysical Journal, vol. 81, no. 3, pp. 1765-1775.
Bajzer Ž, Moncrieffe MCC, Penzar I, Prendergast FG. Complex homogeneous and heterogeneous fluorescence anisotropy decays: Enhancing analysis accuracy. Biophysical Journal. 2001;81(3):1765-1775.
Bajzer, Željko ̌ ; Moncrieffe, M. C C ; Penzar, Ivo ; Prendergast, Franklyn G. / Complex homogeneous and heterogeneous fluorescence anisotropy decays : Enhancing analysis accuracy. In: Biophysical Journal. 2001 ; Vol. 81, No. 3. pp. 1765-1775.
@article{c182ae2ce8de4b239f0af8a2da292219,
title = "Complex homogeneous and heterogeneous fluorescence anisotropy decays: Enhancing analysis accuracy",
abstract = "In biological macromolecules, fluorophores often exhibit multiple depolarizing motions that require multiple lifetimes and rotational relaxation times to define fluorescence intensity and anisotropy decays. The related analysis of time-correlated single-photon counting data becomes uncertain due to the multitude of decay parameters and numerical sensitivity to deconvolution of the instrument response function (IRF) via discretization of integrals. By using simulations we show that improved discretizations based on quadratic and cubic local approximations of the IRF yield more accurate estimation of short rotational relaxation times and lifetimes than the commonly used Grinvald-Steinberg discretization, which in turn appears more reliable than two discretizations based on linear local approximations of the IRF. In addition, our simulation suggests that cubic approximation is the most advantageous in discriminating complex heterogeneous and homogeneous anisotropy decay. We show that among three different information criteria, the Akaike information criterion is best suited for detection of heterogeneity in rotational relaxation times. It is capable of detecting heterogeneity even when anisotropy decay appears homogeneous within statistical errors of estimation.",
author = "Bajzer, {Željko ̌} and Moncrieffe, {M. C C} and Ivo Penzar and Prendergast, {Franklyn G.}",
year = "2001",
language = "English (US)",
volume = "81",
pages = "1765--1775",
journal = "Biophysical Journal",
issn = "0006-3495",
publisher = "Biophysical Society",
number = "3",

}

TY - JOUR

T1 - Complex homogeneous and heterogeneous fluorescence anisotropy decays

T2 - Enhancing analysis accuracy

AU - Bajzer, Željko ̌

AU - Moncrieffe, M. C C

AU - Penzar, Ivo

AU - Prendergast, Franklyn G.

PY - 2001

Y1 - 2001

N2 - In biological macromolecules, fluorophores often exhibit multiple depolarizing motions that require multiple lifetimes and rotational relaxation times to define fluorescence intensity and anisotropy decays. The related analysis of time-correlated single-photon counting data becomes uncertain due to the multitude of decay parameters and numerical sensitivity to deconvolution of the instrument response function (IRF) via discretization of integrals. By using simulations we show that improved discretizations based on quadratic and cubic local approximations of the IRF yield more accurate estimation of short rotational relaxation times and lifetimes than the commonly used Grinvald-Steinberg discretization, which in turn appears more reliable than two discretizations based on linear local approximations of the IRF. In addition, our simulation suggests that cubic approximation is the most advantageous in discriminating complex heterogeneous and homogeneous anisotropy decay. We show that among three different information criteria, the Akaike information criterion is best suited for detection of heterogeneity in rotational relaxation times. It is capable of detecting heterogeneity even when anisotropy decay appears homogeneous within statistical errors of estimation.

AB - In biological macromolecules, fluorophores often exhibit multiple depolarizing motions that require multiple lifetimes and rotational relaxation times to define fluorescence intensity and anisotropy decays. The related analysis of time-correlated single-photon counting data becomes uncertain due to the multitude of decay parameters and numerical sensitivity to deconvolution of the instrument response function (IRF) via discretization of integrals. By using simulations we show that improved discretizations based on quadratic and cubic local approximations of the IRF yield more accurate estimation of short rotational relaxation times and lifetimes than the commonly used Grinvald-Steinberg discretization, which in turn appears more reliable than two discretizations based on linear local approximations of the IRF. In addition, our simulation suggests that cubic approximation is the most advantageous in discriminating complex heterogeneous and homogeneous anisotropy decay. We show that among three different information criteria, the Akaike information criterion is best suited for detection of heterogeneity in rotational relaxation times. It is capable of detecting heterogeneity even when anisotropy decay appears homogeneous within statistical errors of estimation.

UR - http://www.scopus.com/inward/record.url?scp=0034866087&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034866087&partnerID=8YFLogxK

M3 - Article

C2 - 11509386

AN - SCOPUS:0034866087

VL - 81

SP - 1765

EP - 1775

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 3

ER -