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 TokyoHongo 7-3-1,Bunkyo-ku, Tokyo 113-8656,Japan

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

Outline

introduce the device work principle fabrication result conclusion

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 .

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

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

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).

Fabrication

result

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

HUVEC

Antibody

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

Conclusion

Label-free sorting. Higher throughput In the future, We can identify for some our diff

erent bodies cells, such as stem cells or monocyte. It can help us to recognize how it work.