Selection and tuning of a fast and simple phase-contrast microscopy image segmentation algorithm for measuring myoblast growth kinetics in an automated manner

Microsc Microanal. 2013 Aug;19(4):855-66. doi: 10.1017/S143192761300161X. Epub 2013 May 30.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Automation, Laboratory / methods
  • Cell Proliferation*
  • Cells, Cultured
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Microscopy, Phase-Contrast / methods*
  • Myoblasts / cytology*
  • Myoblasts / physiology*