title: Selection and tuning of a fast and simple phase-contrast microscopy image segmentation algorithm for measuring myoblast growth kinetics in an automated manner.
abstract: Acquiring and processing phase-contrast microscopy images in wide-field long-term live-cell imaging and high-throughput screening applications is still a challenge as the methodology and algorithms used must be fast, simple to use and tune, and as minimally intrusive as possible. In this paper, we developed a simple and fast algorithm to compute the cell-covered surface (degree of confluence) in phase-contrast microscopy images. This segmentation algorithm is based on a range filter of a specified size, a minimum range threshold, and a minimum object size threshold. These parameters were adjusted in order to maximize the F-measure function on a calibration set of 200 hand-segmented images, and its performance was compared with other algorithms proposed in the literature. A set of one million images from 37 myoblast cell cultures under different conditions were processed to obtain their cell-covered surface against time. The data were used to fit exponential and logistic models, and the analysis showed a linear relationship between the kinetic parameters and passage number and highlighted the effect of culture medium quality on cell growth kinetics. This algorithm could be used for real-time monitoring of cell cultures and for high-throughput screening experiments upon adequate tuning.
authors.affiliation: Department of Chemical Engineering, Université Laval, Pavillon Adrien-Pouliot, 1065 ave. de la Médecine, Québec City, Québec G1V 0A6, Canada.
authors.forename: Pierre-Marc
authors.initials: PM
authors.lastname: Juneau
authors.forename: Alain
authors.initials: A
authors.lastname: Garnier
authors.forename: Carl
authors.initials: C
authors.lastname: Duchesne
mesh_terms: D000465:Algorithms; D057205:Automation, Laboratory / Q000379:methods; D049109:Cell Proliferation; D002478:Cells, Cultured; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008858:Microscopy, Phase-Contrast / Q000379:methods*; D032446:Myoblasts / Q000166:cytology* / Q000502:physiology*