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 Wun-Hao Wu ( 吳文豪 ) Reporter: Wun-Hao Wu ( 吳文豪 ) 11/21, 2007

2. Outline introduce the device work principle fabrication result conclusion

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

4. μTAS ’05,Boston,MA,USA,Oct,9-13,2005,pp.868-870. dix AM Two advantage of using diX AM for immunoreactive surface. 1.It is provided as-deposited on conformally-coated three dimensional structures. 2.It is highly biocompatible in nature. diX AM substrate

5. μTAS ’05,Boston,MA,USA,Oct,9-13,2005,pp.868-870. principle 1.Adsorbing the Antibodies 2.Cell inject into the microchannel 3.Passing through the Sample plug 4.Then, Passing through the Cell sorting column. device

6. Adsorbing the Antibodies 1.Conjugating NHS-LC-LC-biotin to surface amines. 2.NHS-LC-LC-biotin is dissolved into dimethylsulfoxide. 3.pouring into bicine buffered saline(pH 8). 4.Biotin solution is introduced into micro column and incubated for one hour at 30 ℃. 5.Streptavidin and biotin- conjugated CD31 antibody solution is,each of them dissolved into PBS(pH 7.4).

7. Fabrication

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

9. HUVEC and HL60 are mixed at 5 ×10 5 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.