Cell-Based Microarrays pp 191-206

Part of the Methods in Molecular Biology book series (MIMB, volume 706)

Microfluidic Image Cytometry

  • Ken-ichiro Kamei
  • Jing Sun
  • Hsian-Rong Tseng
  • Robert Damoiseaux
Protocol

Abstract

Cell-based arrays offer powerful tools for genomics/proteomics and drug discovery, and are widely applicable for most cell lines. However, it is challenging to apply cell-based arrays for in vitro diagnosis due to limited amount of patient samples. Here, we utilized and demonstrated microfluidic image cytometry (MIC), capable of quantitative, single-cell profiling of multiple signaling molecules using only 300–3,000 cells from clinical brain tumor specimens for in vitro molecular diagnosis. First, we characterized the PI3K/AKT/mTOR pathway, which is often over-activated in the brain tumors, in U87 brain tumor cell lines by measuring EGFR, PTEN, pAKT, and pS6 with a MIC platform, and applied this measurement to clinical brain tumor specimens. In conjunction with statistical analysis, we were able to characterize extensive inter- and intra-tumoral molecular heterogeneity.

Key words

Microfluidic image cytometry molecular diagnosis glioblastoma Pi3k signaling pathway immunofluorescence biostatistics 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Ken-ichiro Kamei
    • 1
  • Jing Sun
    • 1
  • Hsian-Rong Tseng
    • 1
  • Robert Damoiseaux
    • 2
  1. 1.Department of Molecular & Medical PharmacologyDavid Geffen School of Medicine, University of CaliforniaLos AngelesUSA
  2. 2.Molecular Screening Shared ResourceDavid Geffen School of Medicine, University of CaliforniaLos AngelesUSA

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