title: A quantitative image cytometry technique for time series or population analyses of signaling networks.
abstract: BACKGROUND
  Modeling of cellular functions on the basis of experimental observation is increasingly common in the field of cellular signaling. However, such modeling requires a large amount of quantitative data of signaling events with high spatio-temporal resolution. A novel technique which allows us to obtain such data is needed for systems biology of cellular signaling.
  
  
  RESULTS
  We developed a fully automatable assay technique, termed quantitative image cytometry (QIC), which integrates a quantitative immunostaining technique and a high precision image-processing algorithm for cell identification. With the aid of an automated sample preparation system, this device can quantify protein expression, phosphorylation and localization with subcellular resolution at one-minute intervals. The signaling activities quantified by the assay system showed good correlation with, as well as comparable reproducibility to, western blot analysis. Taking advantage of the high spatio-temporal resolution, we investigated the signaling dynamics of the ERK pathway in PC12 cells.
  
  
  CONCLUSIONS
  The QIC technique appears as a highly quantitative and versatile technique, which can be a convenient replacement for the most conventional techniques including western blot, flow cytometry and live cell imaging. Thus, the QIC technique can be a powerful tool for investigating the systems biology of cellular signaling.
authors.affiliation: Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo, Japan.
authors.forename: Yu-ichi
authors.initials: Y
authors.lastname: Ozaki
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Uda
authors.forename: Takeshi H
authors.initials: TH
authors.lastname: Saito
authors.forename: Jaehoon
authors.initials: J
authors.lastname: Chung
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kubota
authors.forename: Shinya
authors.initials: S
authors.lastname: Kuroda
mesh_terms: D000465:Algorithms; D000818:Animals; D001331:Automation; D048049:Extracellular Signal-Regulated MAP Kinases / Q000032:analysis; D019044:Image Cytometry / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D016716:PC12 Cells; D051381:Rats; D015398:Signal Transduction; D049490:Systems Biology / Q000379:methods*; D013997:Time Factors
chemical_list: D048049:Extracellular Signal-Regulated MAP Kinases