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Analysis and applications of fluores...

Chen, Yan.

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Analysis and applications of fluorescence fluctuation spectroscopy.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Analysis and applications of fluorescence fluctuation spectroscopy./
Author:	Chen, Yan.
Description:	136 p.
Notes:	Source: Dissertation Abstracts International, Volume: 60-09, Section: B, page: 4464.
Contained By:	Dissertation Abstracts International60-09B.
Subject:	Biophysics, General. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9944818
ISBN:	0599466901

Analysis and applications of fluorescence fluctuation spectroscopy.
Chen, Yan.

Analysis and applications of fluorescence fluctuation spectroscopy. - 136 p.

Source: Dissertation Abstracts International, Volume: 60-09, Section: B, page: 4464.

Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1999.

Fluorescence correlation spectroscopy (FCS) exploits the fluctuations of particles to determine kinetic processes from the autocorrelation function and is used to obtain information about the number of fluorescent particles in a small observation volume. We have developed a new technique, the photon counting histogram (PCH) method, to analyze fluctuation data. PCH and autocorrelation analysis focus on different properties of the fluctuation process: the autocorrelation function is a measure of the time-dependent decay of the fluctuations to their equilibrium value, while the photon counting histogram describes the amplitude distribution of these fluctuations. We derived the theory of the PCH for fluorescence fluctuation experiments and constructed an algorithm to calculate the histogram numerically. Measured photon counting distributions obtained with a two-photon excitation source agree within experimental error with the theoretical PCH.

ISBN: 0599466901Subjects--Topical Terms:

1019105
Biophysics, General.

Analysis and applications of fluorescence fluctuation spectroscopy.
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100 1 $a Chen, Yan. $3 1250924
245 1 0 $a Analysis and applications of fluorescence fluctuation spectroscopy.
300 $a 136 p.
500 $a Source: Dissertation Abstracts International, Volume: 60-09, Section: B, page: 4464.
500 $a Adviser: Enrico Gratton.
502 $a Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1999.
520 $a Fluorescence correlation spectroscopy (FCS) exploits the fluctuations of particles to determine kinetic processes from the autocorrelation function and is used to obtain information about the number of fluorescent particles in a small observation volume. We have developed a new technique, the photon counting histogram (PCH) method, to analyze fluctuation data. PCH and autocorrelation analysis focus on different properties of the fluctuation process: the autocorrelation function is a measure of the time-dependent decay of the fluctuations to their equilibrium value, while the photon counting histogram describes the amplitude distribution of these fluctuations. We derived the theory of the PCH for fluorescence fluctuation experiments and constructed an algorithm to calculate the histogram numerically. Measured photon counting distributions obtained with a two-photon excitation source agree within experimental error with the theoretical PCH.
520 $a We revived another analysis method, moment analysis, which was introduced almost ten years ago. This method calculates G(0) from the fluctuation moments of the photon counts and corrects for their shot noise contribution. The main advantage of moment analysis is its simplicity and model independent determination of G(0).
520 $a Binary dye mixtures were used to demonstrate the influence of the molecular brightness on the fluctuation amplitude G(0). This dependence allows to resolve multiple species based on the difference in their molecular brightness, which we applied to a titration study of an antibody with its ligand. The global fit of the average photon counts &lang;k&rang; and the fluctuation amplitude G(0) as a function of the protein concentration determined the number of binding sites and the dissociation coefficient KD. The analysis further demonstrated heterogeneity of the antibody complex on a time scale not easily accessible by other techniques.
590 $a School code: 0090.
650 4 $a Biophysics, General. $3 1019105
650 4 $a Statistics. $3 517247
690 $a 0786
690 $a 0463
710 2 0 $a University of Illinois at Urbana-Champaign. $3 626646
773 0 $t Dissertation Abstracts International $g 60-09B.
790 1 0 $a Gratton, Enrico, $e advisor
790 $a 0090
791 $a Ph.D.
792 $a 1999
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9944818