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

摘要

Abstract: We have studied three-dimensional reconstructionmethods to estimate the cell volume of astroglial cellsin primary culture. The studies are based onfluorescence imaging and optical sectioning. Anautomated image-acquisition system was developed tocollect two-dimensional microscopic images. Images werereconstructed by the Linear Maximum a Posteriori methodand the non-linear Maximum Likelihood ExpectationMaximization (ML-EM) method. In addition, because ofthe high computational demand of the ML-EM algorithm,we have developed a fast variant of this method. (1)Advanced image analysis techniques were applied foraccurate and automated cell volume determination. (2)The sensitivity and accuracy of the reconstructionmethods were evaluated by using fluorescent micro-beadswith known diameter. The algorithms were applied tofura-2-labeled astroglial cells in primary cultureexposed to hypo- or hyper-osmotic stress. The resultsshowed that the ML-EM reconstructed images are adequatefor the determination of volume changes in cells orparts thereof. !22