Immunomagnetic T cell capture from blood for PCR analysis using microfluidic systems

被引:124
作者
Furdui, VI [1 ]
Harrison, DJ [1 ]
机构
[1] Univ Alberta, Dept Chem, Edmonton, AB T6G 2G2, Canada
关键词
D O I
10.1039/b409366f
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A one-step immunomagnetic separation technique was performed on a microfluidic platform for the isolation of specific cells from blood samples. The cell isolation and purification studies targeted T cells, as a model for low abundance cells ( about 1: 10,000 cells), with more dilute cells as the ultimate goal. T cells were successfully separated on-chip from human blood and from reconstituted blood samples. Quantitative polymerase chain reaction analysis of the captured cells was used to characterize the efficiency of T cell capture in a variety of flow path designs. Employing many (4-8), 50 mm deep narrow channels, with the same overall cross section as a single, 3 mm wide channel, was much more effective in structuring dense enough magnetic bead beds to trap cells in a flowing stream. The use of 8-multiple bifurcated flow paths increased capture efficiencies from similar to20 up to 37%, when compared to a straight 8-way split design, indicating the value of ensuring uniform flow distribution into each channel in a flow manifold for effective cell capture. Sample flow rates of up to 3 muL min(-1) were evaluated in these capture beds.
引用
收藏
页码:614 / 618
页数:5
相关论文
共 29 条
[1]   Purification and characterization of PCR-inhibitory components in blood cells [J].
Abu al-Soud, W ;
Rådström, P .
JOURNAL OF CLINICAL MICROBIOLOGY, 2001, 39 (02) :485-493
[2]   A fully integrated micromachined magnetic particle separator [J].
Ahn, CH ;
Allen, MG ;
Trimmer, W ;
Jun, YN ;
Erramilli, S .
JOURNAL OF MICROELECTROMECHANICAL SYSTEMS, 1996, 5 (03) :151-158
[3]  
AKANE A, 1994, J FORENSIC SCI, V39, P362
[4]   Micro total analysis systems. 2. Analytical standard operations and applications [J].
Auroux, PA ;
Iossifidis, D ;
Reyes, DR ;
Manz, A .
ANALYTICAL CHEMISTRY, 2002, 74 (12) :2637-2652
[5]   Preparation and hybridization analysis of DNA/RNA from E-coli on microfabricated bioelectronic chips [J].
Cheng, J ;
Sheldon, EL ;
Wu, L ;
Uribe, A ;
Gerrue, LO ;
Carrino, J ;
Heller, MJ ;
O'Connell, JP .
NATURE BIOTECHNOLOGY, 1998, 16 (06) :541-546
[6]   Chip PCR .2. Investigation of different PCR amplification systems in microfabricated silicon-glass chips [J].
Cheng, J ;
Shoffner, MA ;
Hvichia, GE ;
Kricka, LJ ;
Wilding, P .
NUCLEIC ACIDS RESEARCH, 1996, 24 (02) :380-385
[7]  
Chhanabhai M, 1997, AM J CLIN PATHOL, V108, P295
[8]  
Chiem NH, 1998, CLIN CHEM, V44, P591
[9]   FIELD AMPLIFIED SAMPLE INJECTION IN HIGH-PERFORMANCE CAPILLARY ELECTROPHORESIS [J].
CHIEN, RL ;
BURGI, DS .
JOURNAL OF CHROMATOGRAPHY, 1991, 559 (1-2) :141-152
[10]   Integrated chip-based capillary electrophoresis [J].
Effenhauser, CS ;
Bruin, GJM ;
Paulus, A .
ELECTROPHORESIS, 1997, 18 (12-13) :2203-2213