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Clinical microfluidics for neutrophil genomics and proteomics.
Nat Med. 2010 Aug 29;
Authors: Kotz KT, Xiao W, Miller-Graziano C, Qian WJ, Russom A, Warner EA, Moldawer LL, De A, Bankey PE, Petritis BO, Camp DG, Rosenbach AE, Goverman J, Fagan SP, Brownstein BH, Irimia D, Xu W, Wilhelmy J, Mindrinos MN, Smith RD, Davis RW, Tompkins RG, Toner M, , Baker HV, Balis UG, Billiar TR, Calvano SE, Cobb JP, Cuschieri J, Finnerty CC, Gamelli RL, Gibran NS, Harbrecht BG, Hayden DL, Hennessy L, Herndon DN, Jeschke MG, Johnson JL, Klein MB, Lowry SF, Maier RV, Mason PH, McDonald-Smith GP, Minei JP, Moore EE, Nathens AB, O Keefe GE, Rahme LG, Remick DG, Schoenfeld DA, Shapiro MB, Sperry J, Storey JD, Tibshirani R, Warren HS, West MA, Wispelwey B, Wong WH
Neutrophils have key roles in modulating the immune response. We present a robust methodology for rapidly isolating neutrophils directly from whole blood with ‘on-chip’ processing for mRNA and protein isolation for genomics and proteomics. We validate this device with an ex vivo stimulation experiment and by comparison with standard bulk isolation methodologies. Last, we implement this tool as part of a near-patient blood processing system within a multi-center clinical study of the immune response to severe trauma and burn injury. The preliminary results from a small cohort of subjects in our study and healthy controls show a unique time-dependent gene expression pattern clearly demonstrating the ability of this tool to discriminate temporal transcriptional events of neutrophils within a clinical setting.
PMID: 20802500 [PubMed - as supplied by publisher]