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Simple method for quantification of fast plasma-membrane movements

Nicolas Van Larebeke (UGent) , Marc Bracke (UGent) and Marcus Mareel (UGent)
(1992) CYTOMETRY. 13(1). p.1-8
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Abstract
We present a method for the quantification of the fast plasma membrane movements that are involved in ruffling, blebbing, fast shape change, and fast translocation. The method is based on the Kontron Vidas image analysis computer program. Video images from cells viewed through an inverted microscope were transmitted to the computer. The procedure was as follows: 4 consecutive video images were averaged (image1); 28 s later a second set of 4 video images was averaged (image 2); image 2 was subtracted from image 1 and the grey level of each pixel of the resulting image was increased with 128 grey level units, resulting in the subtraction image, showing a uniform grey background speckled with brighter and darker spots corresponding to areas of movement. These spots were discriminated and turned into white objects against a black background. Interactive editing was used to delete artefacts that resulted from floating debris. The total area of the discriminated objects was measured, and the parameter motile area in mu-m2 per cell was calculated. We have applied our method to the study of motility induced in epithelial cell lines by the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate and by epidermal growth factor.
Keywords
CELL MEMBRANE, CELL MOTILITY, CELL-SURFACE, RUFFLING, BLEBBING, CELL-SHAPE, TRANSLOCATION, IMAGE ANALYSIS, TPA, EGF, GROWTH-FACTOR, CULTURE, REORGANIZATION, FIBROBLASTS, INDUCTION, CARCINOMA, INVASION, INVITRO, ACTIN

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Citation

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Chicago
Van Larebeke, Nicolas, Marc Bracke, and Marcus Mareel. 1992. “Simple Method for Quantification of Fast Plasma-membrane Movements.” Cytometry 13 (1): 1–8.
APA
Van Larebeke, N., Bracke, M., & Mareel, M. (1992). Simple method for quantification of fast plasma-membrane movements. CYTOMETRY, 13(1), 1–8.
Vancouver
1.
Van Larebeke N, Bracke M, Mareel M. Simple method for quantification of fast plasma-membrane movements. CYTOMETRY. 1992;13(1):1–8.
MLA
Van Larebeke, Nicolas, Marc Bracke, and Marcus Mareel. “Simple Method for Quantification of Fast Plasma-membrane Movements.” CYTOMETRY 13.1 (1992): 1–8. Print.
@article{151151,
  abstract     = {We present a method for the quantification of the fast plasma membrane movements that are involved in ruffling, blebbing, fast shape change, and fast translocation. The method is based on the Kontron Vidas image analysis computer program. Video images from cells viewed through an inverted microscope were transmitted to the computer. The procedure was as follows: 4 consecutive video images were averaged (image1); 28 s later a second set of 4 video images was averaged (image 2); image 2 was subtracted from image 1 and the grey level of each pixel of the resulting image was increased with 128 grey level units, resulting in the subtraction image, showing a uniform grey background speckled with brighter and darker spots corresponding to areas of movement. These spots were discriminated and turned into white objects against a black background. Interactive editing was used to delete artefacts that resulted from floating debris. The total area of the discriminated objects was measured, and the parameter motile area in mu-m2 per cell was calculated. We have applied our method to the study of motility induced in epithelial cell lines by the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate and by epidermal growth factor.},
  author       = {Van Larebeke, Nicolas and Bracke, Marc and Mareel, Marcus},
  issn         = {0196-4763},
  journal      = {CYTOMETRY},
  keyword      = {CELL MEMBRANE,CELL MOTILITY,CELL-SURFACE,RUFFLING,BLEBBING,CELL-SHAPE,TRANSLOCATION,IMAGE ANALYSIS,TPA,EGF,GROWTH-FACTOR,CULTURE,REORGANIZATION,FIBROBLASTS,INDUCTION,CARCINOMA,INVASION,INVITRO,ACTIN},
  language     = {eng},
  number       = {1},
  pages        = {1--8},
  title        = {Simple method for quantification of fast plasma-membrane movements},
  url          = {http://dx.doi.org/10.1002/cyto.990130103},
  volume       = {13},
  year         = {1992},
}

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