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A method for quantifying cell size from differential interference contrast images: validation and application to osmotically stressed chondrocytes
Author(s) -
Alexopoulos L. G.,
Erickson G. R.,
Guilak F.
Publication year - 2002
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1046/j.0022-2720.2001.00976.x
Subject(s) - differential interference contrast microscopy , enhanced data rates for gsm evolution , contrast (vision) , interference (communication) , optics , microscopy , biological system , range (aeronautics) , confocal microscopy , edge detection , volume (thermodynamics) , computer science , materials science , image processing , artificial intelligence , physics , image (mathematics) , biology , computer network , channel (broadcasting) , quantum mechanics , composite material
Summary An automatic image analysis method was developed to determine the shape and size of spheroidal cells from a time series of differential interference contrast (DIC) images. The program incorporates an edge detection algorithm and dynamic programming for edge linking. To assess the accuracy and working range of the method, results from DIC images of different focal planes and resolutions were compared to confocal images in which the cell membrane was fluorescently labelled. The results indicate that a 1‐µm focal drift from the in‐focus plane can lead to an overestimation of cell volume up to 14.1%, mostly due to shadowing effects of DIC microscopy. DIC images allow for accurate measurements when the focal plane lies in a zone slightly above the centre of a spherical cell. In this range the method performs with 1.9% overall volume error without taking into account the error introduced by the representation of the cell as a sphere. As a test case, the method was applied to quantify volume changes due to acute changes of osmotic stress.