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Proceedings Paper

Quantifying fluorescence signals in confocal image stacks deep in turbid media
Author(s): S. Beer; U. Maeder; T. Bergmann; J. M. Burg; M. Fiebich; F. Runkel
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Paper Abstract

When confocal depth stacks are taken, the collected signal (normally the fluorescence signal), decays dependent of the depth of the confocal slice in the turbid medium. This decay is caused by scattering and absorption of the exciting light and of the fluorescence light. As the attenuation parameters, i.e. scattering and absorption coefficients, are normally unknown when observing a new sample, a method is proposed to compensate for the attenuation of the involved light by correcting the fluorescence signal using the attenuation behavior of the sample measured directly on the spot where the fluorescence stack is taken. The method works without any a priori knowledge about the optical properties of the sample. Using this self-reference technique, a confocal fluorescence depth stack can be created where the signal intensity is not dependent on the scattering and absorption caused intensity decay. The proposed method is tested on fluorescent beads embedded in scattering and absorbing hydrogel phantoms.

Paper Details

Date Published: 2 February 2012
PDF: 6 pages
Proc. SPIE 8227, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIX, 82271G (2 February 2012); doi: 10.1117/12.907875
Show Author Affiliations
S. Beer, Technische Hochschule Mittelhessen Univ. of Applied Sciences Giessen (Germany)
U. Maeder, Technische Hochschule Mittelhessen Univ. of Applied Sciences Giessen (Germany)
T. Bergmann, Technische Hochschule Mittelhessen Univ. of Applied Sciences Giessen (Germany)
J. M. Burg, Technische Hochschule Mittelhessen Univ. of Applied Sciences Giessen (Germany)
M. Fiebich, Technische Hochschule Mittelhessen Univ. of Applied Sciences Giessen (Germany)
F. Runkel, Technische Hochschule Mittelhessen Univ. of Applied Sciences Giessen (Germany)


Published in SPIE Proceedings Vol. 8227:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIX
Jose-Angel Conchello; Carol J. Cogswell; Tony Wilson; Thomas G. Brown, Editor(s)

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