Single-cell volume estimation by applying three-dimensional reconstruction methods

Siamak Khatibi; Louise Allansson; Tomas Gustavsson; Fredrik Blomstrand; Elisabeth Hansson; Torsten Olsson

doi:10.1117/12.347561

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6 May 1999 Single-cell volume estimation by applying three-dimensional reconstruction methods

Siamak Khatibi, Louise Allansson, Tomas Gustavsson, Fredrik Blomstrand, Elisabeth Hansson, Torsten Olsson

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Proceedings Volume 3605, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing VI; (1999) https://doi.org/10.1117/12.347561
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

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.

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Siamak Khatibi, Louise Allansson, Tomas Gustavsson, Fredrik Blomstrand, Elisabeth Hansson, and Torsten Olsson "Single-cell volume estimation by applying three-dimensional reconstruction methods", Proc. SPIE 3605, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing VI, (6 May 1999); https://doi.org/10.1117/12.347561

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