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Proceedings Paper

Single-cell volume estimation by applying three-dimensional reconstruction methods
Author(s): Siamak Khatibi; Louise Allansson; Tomas Gustavsson; Fredrik Blomstrand; Elisabeth Hansson; Torsten Olsson
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Paper Abstract

We have studied three-dimensional reconstruction methods to estimate the cell volume of astroglial cells in primary culture. The studies are based on fluorescence imaging and optical sectioning. An automated image-acquisition system was developed to collect two-dimensional microscopic images. Images were reconstructed by the Linear Maximum a Posteriori method and the non-linear Maximum Likelihood Expectation Maximization (ML-EM) method. In addition, because of the high computational demand of the ML-EM algorithm, we have developed a fast variant of this method. (1) Advanced image analysis techniques were applied for accurate and automated cell volume determination. (2) The sensitivity and accuracy of the reconstruction methods were evaluated by using fluorescent micro-beads with known diameter. The algorithms were applied to fura-2-labeled astroglial cells in primary culture exposed to hypo- or hyper-osmotic stress. The results showed that the ML-EM reconstructed images are adequate for the determination of volume changes in cells or parts thereof.

Paper Details

Date Published: 6 May 1999
PDF: 12 pages
Proc. SPIE 3605, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing VI, (6 May 1999); doi: 10.1117/12.347561
Show Author Affiliations
Siamak Khatibi, Chalmers Univ. of Technology (Sweden)
Louise Allansson, Goeteborg Univ. (Sweden)
Tomas Gustavsson, Chalmers Univ. of Technology (Sweden)
Fredrik Blomstrand, Goeteborg Univ. (Sweden)
Elisabeth Hansson, Goeteborg Univ. (Sweden)
Torsten Olsson, Chalmers Univ. of Technology (Sweden)

Published in SPIE Proceedings Vol. 3605:
Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing VI
Dario Cabib; Carol J. Cogswell; Jose-Angel Conchello; Jeremy M. Lerner; Tony Wilson, Editor(s)

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