title: Microfluidic image cytometry.
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.
authors.affiliation: Department of Molecular & Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, CA, USA. kkamei@icems.kyoto-u.ac.jp
authors.forename: Ken-ichiro
authors.initials: K
authors.lastname: Kamei
authors.forename: Jing
authors.initials: J
authors.lastname: Sun
authors.forename: Hsian-Rong
authors.initials: HR
authors.lastname: Tseng
authors.forename: Robert
authors.initials: R
authors.lastname: Damoiseaux
mesh_terms: D001932:Brain Neoplasms / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000032:analysis / Q000378:metabolism; D005909:Glioblastoma / Q000378:metabolism / Q000473:pathology; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D007150:Immunohistochemistry; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence; D051059:PTEN Phosphohydrolase / Q000032:analysis / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000032:analysis / Q000378:metabolism; D015203:Reproducibility of Results; D019893:Ribosomal Protein S6 Kinases / Q000032:analysis / Q000378:metabolism; D046888:Tissue Array Analysis / Q000379:methods
chemical_list: D066246:ErbB Receptors; D047908:Intracellular Signaling Peptides and Proteins; D051059:PTEN Phosphohydrolase; D019869:Phosphatidylinositol 3-Kinases; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D019893:Ribosomal Protein S6 Kinases; C512478:EGFR protein, human; C494929:PTEN protein, human