title: Determination of single cell surface protein expression using a tagless microfluidic method.
abstract: We describe a method to detect the expression of a surface protein in single cells without prior labeling or manipulation using a microfluidic device. When the protein is expressed on a cell surface, it undergoes transient bond formation with an immobilized ligand as the cell is pumped through a microfluidic channel, resulting in a specific decrease in the cell's velocity. We were able to detect the expression of interleukin 13 receptor alpha 2 (IL13Rα2) differentially expressed on LM2 cells, a subline of MDA-MB-231 human breast cancer cells with a unique lung metastatic capability. The detection of cells with high expression of the protein was near 100% sensitive and 100% specific. We also provided proof of principle of multiplexing by tracking the same cell over two, differentially-coated patches. The method is non-destructive and cells can be collected for reanalysis. The system can identify positive cells in a cell mixture. This method will have a potential impact in analyzing cancer cells when only a few are available, such as the case with needle aspirates and when labeling and manipulation result in cell loss.
authors.affiliation: Genome Data Systems, Inc., Hamilton, NJ, USA.
authors.forename: Rajan
authors.initials: R
authors.lastname: Kumar
authors.forename: Sri HariKrishna
authors.initials: SH
authors.lastname: Vellanki
authors.forename: Robin
authors.initials: R
authors.lastname: Smith
authors.forename: Robert
authors.initials: R
authors.lastname: Wieder
mesh_terms: D045744:Cell Line, Tumor; D005434:Flow Cytometry / Q000295:instrumentation* / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D053663:Interleukin-13 Receptor alpha2 Subunit / Q000032:analysis* / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D059010:Single-Cell Analysis / Q000295:instrumentation* / Q000379:methods*
chemical_list: D053663:Interleukin-13 Receptor alpha2 Subunit