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Proceedings Paper

Analysis of ultrasensitive fluorescence experiments
Author(s): Yuxing Sun; Bruce A. Whitehead; Lloyd M. Davis
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Paper Abstract

DNA sequencing and several other applications of single- molecule detection (SMD) currently under development utilize spectroscopic measurements for categorization of different types of fluorophores. In the collection and analysis of data from such experiments, the photon signals are sorted into different channels, depending upon their arrival time, emission wavelength, or other distinguishable properties. If the photon statistics are adequate, maximum-likelihood estimation (MLE) techniques can be successfully applied to determine which fluorophore is present. However, data analysis using neural network (NN) methods can offer several advantages. We consider data from a Monte Carlo simulation of SMD in a flow-cell, in which a time-resolved fluorescence decay profile is accumulated for each photon burst. A 2-layer NN, with sigmoid as the activation function, is trained on a set of simulated data using back-propagation and the (delta) - learning rule, and then used for identification of photon bursts in subsequent simulations. The NN is able to consider additional input parameters, such as the amplitudes of the weighted-sliding-sum digital-filter output of the photon bursts and the durations of the bursts. It can yield superior identification of photon bursts, particularly in cases where the fluorophores have disparate fluorescence quantum efficiencies, absorption cross-sections, or photodegradation efficiencies, or where the categorization includes other possibilities, such as background fluctuations, or the simultaneous presence of both fluorophores.

Paper Details

Date Published: 3 May 1999
PDF: 12 pages
Proc. SPIE 3602, Advances in Fluorescence Sensing Technology IV, (3 May 1999); doi: 10.1117/12.347537
Show Author Affiliations
Yuxing Sun, Univ. of Tennessee Space Institute (United States)
Bruce A. Whitehead, Univ. of Tennessee Space Institute (United States)
Lloyd M. Davis, Univ. of Tennessee Space Institute (United States)

Published in SPIE Proceedings Vol. 3602:
Advances in Fluorescence Sensing Technology IV
Joseph R. Lakowicz; Steven A. Soper; Richard B. Thompson, Editor(s)

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