3-DIMENSIONAL LABELING PROGRAM FOR ELUCIDATION OF THE GEOMETRIC-PROPERTIES OF BIOLOGICAL PARTICLES IN 3-DIMENSIONAL SPACE

For all biological particles such as cells or cellular organelles, the re are three-dimensional coordinates representing the centroid or cent er of gravity, These coordinates and other numerical parameters such a s volume, fluorescence intensity, surface area, and shape are referred to in this paper as geometric properties, which may provide critical information for the clarification of in situ mechanisms of molecular a nd cellular functions in living organisms, We have established a metho d for the elucidation of these properties, designated the three-dimens ional labeling program (3DLP), Algorithms of 3DLP are so simple that t his method can be carried out through the use of software combinations in image analysis on a personal computer, To evaluate 3DLP,it was app lied to a 32-cell-stage sea urchin embryo, double stained with FITC fo r cellular protein of blastomeres and propidium iodide for nuclear DNA , A stack of optical serial section images was obtained by confocal la ser scanning microscopy, The method was found effective for determinin g geometric properties and should prove applicable to the study of man y different kinds of biological particles in three-dimensional space. (C) 1996 Academic Press, Inc.