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

Yu Ichi Ozaki, Shinsuke Uda, Takeshi H. Saito, Jaehoon Chung, Hiroyuki Kubota, Shinya Kuroda

Research output: Contribution to journalArticle

23 Citations (Scopus)

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.

Original languageEnglish
Article numbere9955
JournalPloS one
Volume5
Issue number4
DOIs
Publication statusPublished - Sep 10 2010
Externally publishedYes

Fingerprint

Image Cytometry
Cell signaling
Time series
time series analysis
Systems Biology
cell communication
Assays
Western Blotting
Population
Phosphorylation
Flow cytometry
MAP Kinase Signaling System
PC12 Cells
quantitative analysis
Flow Cytometry
Western blotting
Image processing
methodology
Observation
image analysis

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

A quantitative image cytometry technique for time series or population analyses of signaling networks. / Ozaki, Yu Ichi; Uda, Shinsuke; Saito, Takeshi H.; Chung, Jaehoon; Kubota, Hiroyuki; Kuroda, Shinya.

In: PloS one, Vol. 5, No. 4, e9955, 10.09.2010.

Research output: Contribution to journalArticle

Ozaki, Yu Ichi ; Uda, Shinsuke ; Saito, Takeshi H. ; Chung, Jaehoon ; Kubota, Hiroyuki ; Kuroda, Shinya. / A quantitative image cytometry technique for time series or population analyses of signaling networks. In: PloS one. 2010 ; Vol. 5, No. 4.
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