Browsing by Subject "fractionation"
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Item Antibody-Independent Isolation of Circulating Tumor Cells by Continuous-Flow Dielectrophoresis(2013-01) Shim, Sangjo; Stemke-Hale, Katherine; Tsimberidou, Apostolia M.; Noshari, Jamileh; Anderson, Thomas E.; Gascoyne, Peter R. C.; Shim, Sangjo; Noshari, Jamileh; Anderson, Thomas E.; Gascoyne, Peter R. C.Circulating tumor cells (CTCs) are prognostic markers for the recurrence of cancer and may carry molecular information relevant to cancer diagnosis. Dielectrophoresis (DEP) has been proposed as a molecular marker-independent approach for isolating CTCs from blood and has been shown to be broadly applicable to different types of cancers. However, existing batch-mode microfluidic DEP methods have been unable to process 10 ml clinical blood specimens rapidly enough. To achieve the required processing rates of 106 nucleated cells/min, we describe a continuous flow microfluidic processing chamber into which the peripheral blood mononuclear cell fraction of a clinical specimen is slowly injected, deionized by diffusion, and then subjected to a balance of DEP, sedimentation and hydrodynamic lift forces. These forces cause tumor cells to be transported close to the floor of the chamber, while blood cells are carried about three cell diameters above them. The tumor cells are isolated by skimming them from the bottom of the chamber while the blood cells flow to waste. The principles, design, and modeling of the continuous-flow system are presented. To illustrate operation of the technology, we demonstrate the isolation of circulating colon tumor cells from clinical specimens and verify the tumor origin of these cells by molecular analysis. (C) 2013 American Institute of Physics. [http://dx.doi.org/10.1063/1.4774304]Item Interstellar CN And CH+ In Diffuse Molecular Clouds: C-12/C-13 Ratios And Cn Excitation(2011-02) Ritchey, A. M.; Federman, S. R.; Lambert, David L.; Ritchey, A. M.; Federman, S. R.; Lambert, David L.We present very high signal-to-noise ratio absorption-line observations of CN and CH+ along 13 lines of sight through diffuse molecular clouds. The data are examined to extract precise isotopologic ratios of (CN)-C-12/(CN)-C-13 and (CH+)-C-12/(CH+)-C-13 in order to assess predictions of diffuse cloud chemistry. Our results on (CH+)-C-12/(CH+)-C-13 confirm that this ratio does not deviate from the ambient C-12/C-13 ratio in local interstellar clouds, as expected if the formation of CH+ involves nonthermal processes. We find that (CN)-C-12/(CN)-C-13, however, can be significantly fractionated away from the ambient value. The dispersion in our sample of (CN)-C-12/(CN)-C-13 ratios is similar to that found in recent surveys of (CO)-C-12/(CO)-C-13. For sight lines where both ratios have been determined, the (CN)-C-12/(CN)-C-13 ratios are generally fractionated in the opposite sense compared to (CO)-C-12/(CO)-C-13. Chemical fractionation in CO results from competition between selective photodissociation and isotopic charge exchange (ICE). An inverse relationship between (CN)-C-12/(CN)-C-13 and (CO)-C-12/(CO)-C-13 follows from the coexistence of CN and CO in diffuse cloud cores. However, an ICE reaction with CN may mitigate the enhancements in (CN)-C-12/(CN)-C-13 for lines of sight with low (CO)-C-12/(CO)-C-13 ratios. For two sight lines with high values of (CO)-C-12/(CO)-C-13, our results indicate that about 50% of the carbon is locked up in CO, which is consistent with the notion that these sight lines probe molecular cloud envelopes where the transition from C+ to CO is expected to occur. An analysis of CN rotational excitation yields a weighted mean value for T-01((CN)-C-12) of 2.754 +/- 0.002 K, which implies an excess over the temperature of the cosmic microwave background (CMB) of only 29 +/- 3 mK. This modest excess eliminates the need for a local excitation mechanism beyond electron and neutral collisions. The rotational excitation temperatures in (CN)-C-13 show no excess over the temperature of the CMB.Item Letter to H.B. Stenzel from Karl M. Wilbur on 1961-09-14(1961-09-14) Wilbur, Karl M.Item Microfluidic Enrichment of Small Proteins from Complex Biological Mixture on Nanoporous Silica Chip(2011-03) Hu, Ye; Gopal, Ashwini; Lin, Kevin; Peng, Yang; Tasciotti, Ennio; Zhang, Xiojing John; Ferrari, Mauro; Gopal, Ashwini; Lin, Kevin; Zhang, Xiojing JohnThe growing field of miniaturized diagnostics is hindered by a lack of pre-analysis treatments that are capable of processing small sample volumes for the detection of low concentration analytes in a high-throughput manner. This letter presents a novel, highly efficient method for the extraction of low-molecular weight (LMW) proteins from biological fluids, represented by a mixture of standard proteins, using integrated microfluidic systems. We bound a polydimethylsiloxane layer patterned with a microfluidic channel onto a well-defined nanoporous silica substrate. Using rapid, pressure-driven fractionation steps, this system utilizes the size-exclusion properties of the silica nanopores to remove high molecular weight proteins while simultaneously isolating and enriching LMW proteins present in the biological sample. The introduction of the microfluidic component offers important advantages such as high reproducibility, a simple user interface, controlled environment, the ability to process small sample volumes, and precise quantification. This solution streamlines high-throughput proteomics research on many fronts and may find broad acceptance and application in clinical diagnostics and point of care detection. (C) 2011 American Institute of Physics. [doi: 10.1063/1.3528237]