A quantitative image cytometry technique for time series or population analyses of signaling networks

PLoS One. 2010 Apr 1;5(4):e9955. doi: 10.1371/journal.pone.0009955.

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.

Methodology/principal findings: 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/significance: 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Animals
  • Automation
  • Extracellular Signal-Regulated MAP Kinases / analysis
  • Image Cytometry / methods*
  • Image Processing, Computer-Assisted
  • Immunohistochemistry
  • PC12 Cells
  • Rats
  • Signal Transduction*
  • Systems Biology / methods*
  • Time Factors

Substances

  • Extracellular Signal-Regulated MAP Kinases