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Adhesion-Based Cell Sorter with Antibody-immobilized Functionalized-Paryene Surface

Adhesion-Based Cell Sorter with Antibody-immobilized Functionalized-Paryene Surface. Author ： Junichi Miwa, Yuji Suzuki and Nobuhide asagi Department of Mechanical Engineering,The University of Tokyo Hongo 7-3-1,Bunkyo-ku, Tokyo 113-8656,Japan Reporter: Wun-Hao Wu ( 吳文豪 )

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Adhesion-Based Cell Sorter with Antibody-immobilized Functionalized-Paryene Surface

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  1. Adhesion-Based Cell Sorter with Antibody-immobilized Functionalized-Paryene Surface Author：Junichi Miwa, Yuji Suzuki and Nobuhide asagi Department of Mechanical Engineering,The University of Tokyo Hongo 7-3-1,Bunkyo-ku, Tokyo 113-8656,Japan Reporter:Wun-Hao Wu (吳文豪) 11/21, 2007

  2. Outline • introduce • the device work principle • fabrication • result • conclusion

  3. introduce Nature Biotech., vol.17 , no.11,pp.1109-1111.1999 BIOLOGY OF REPRODUCTION 61, 582–589 (1999) The cell sorter is more merit of label-free sorting and the potential for higher throughput .

  4. diX AM substrate dix AM μTAS ’05 ,Boston ,MA,USA ,Oct,9-13,2005,pp.868-870. • Two advantage of using diX AM for immunoreactive surface . • It is provided as-deposited on conformally-coated three dimensional structures. • It is highly biocompatible in nature.

  5. principle device μTAS ’05 ,Boston ,MA,USA ,Oct,9-13,2005,pp.868-870. • Adsorbing the Antibodies • Cell inject into the microchannel • Passing through the Sample plug • Then, Passing through the Cell sorting column.

  6. Adsorbing the Antibodies • Conjugating NHS-LC-LC-biotin to surface amines. • NHS-LC-LC-biotin is dissolved into dimethylsulfoxide. • pouring into bicine buffered saline(pH 8). • Biotin solution is introduced into micro column and incubated for one hour at 30 ℃. • Streptavidin and biotin-conjugated CD31 antibody solution is,each of them dissolved into PBS(pH 7.4).

  7. Fabrication

  8. result HUVEC Antibody Time-averaged HUVEC velocity in microcolumns with various surface antibody immobilizations.

  9. HUVEC and HL60 are mixed at 5 ×105 cells/mL total concentration, at the ratio of 1:1. The flow is driven by a syringe pump at the total flow rate of 1.2 μL/min, with the volume ratio between the main inlet and two hydeodynamic focousing buffer inlets being 2:1:1. the corresponding bulk mean flow velocity in the cell sorting column is 1 mm/s

  10. Conclusion • Label-free sorting. • Higher throughput • In the future, We can identify for some our different bodies cells, such as stem cells or monocyte. It can help us to recognize how it work.

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