A Case Study on 3-D Reconstruction and Shape Description of Peroxisomes in Yeast

Subcellular organelles are commonly analyzed using 2-D fluorescent microscopy. However, 3-D reconstruction and analysis of organelle topology in a high-throughput manner promises to result in a better understanding of cellular systems. We developed image analysis methods for automated quantitative analysis of peroxisome shapes. The methods employ 3-D image stacks obtained by confocal microscopy. There are three fundamental phases: image preprocessing and segmentation, 3-D reconstruction, and automated quantification of peroxisome topology in 3-D using shape descriptors. The algorithms are shown to produce results that can be used to classify objects of different topologies, and to enable visual studies of peroxisomes in 3-D.