Open AccessComparison of several digital and stereological methods for estimating surface area and volume of cells studied by confocal microscopy
Author(s) -
Kubínová Lucie,
Janáček Jiří,
Guilak Farshid,
Opatrný Zdeněk
Publication year - 1999
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/(sici)1097-0320(19990601)36:2<85::aid-cyto1>3.0.co;2-3
Subject(s) - volume (thermodynamics) , stereology , computer science , confocal , triangulation , surface (topology) , contouring , microscope , computer vision , confocal microscopy , microscopy , biomedical engineering , computer graphics (images) , optics , mathematics , physics , geometry , pathology , quantum mechanics , medicine
Background: The implementation of different methods for estimating the surface area and volume of cells studied by confocal microscopy was developed. The methods were compared from the point of view of their precision, applicability and efficiency. Methods: Interactive stereological methods (spatial grid method, fakir method, Cavalieri principle) as well as automatic digital methods (digital Crofton method, voxel counting, triangulation method, iso‐intensity contouring method) were considered. The methods were tested on model geometrical solids and on real volume images consisting of a stack of serial sections encompassing entire tobacco BY‐2 cells or cell chains. Results: It is shown that many of the studied methods are very precise when applied to cells of simple or moderately complex shapes. The automatic digital methods are fast and precise but their applicability is limited by the necessity to segment automatically the object surface and to find an optimal resolution. This limitation is not present in stereological methods which are applied interactively and thus are more time‐consuming. Conclusions: The presented implementations of the fakir method and the Cavalieri principle enable interactive, unbiased and efficient estimation of the cell surface area and volume. The recommended steps for measuring the surface area and/or volume of objects studied by confocal microscopy are described. Cytometry 36:85–95, 1999. © 1999 Wiley‐Liss, Inc.