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Sensors 2013, 13(4), 4674-4693; doi:10.3390/s130404674

Microflow Cytometers with Integrated Hydrodynamic Focusing

1,* , 2
1, 1
1 Physikalisch-Technische Bundesanstalt (PTB), Abbestrasse 2-12, 10587 Berlin, Germany 2 Technische Universität Berlin, Strasse des 17. Juni 135, 10623 Berlin, Germany
* Author to whom correspondence should be addressed.
Received: 1 March 2013 / Revised: 22 March 2013 / Accepted: 28 March 2013 / Published: 9 April 2013
(This article belongs to the Special Issue Microfluidic Devices)
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This study demonstrates the suitability of microfluidic structures for high throughput blood cell analysis. The microfluidic chips exploit fully integrated hydrodynamic focusing based on two different concepts: Two-stage cascade focusing and spin focusing (vortex) principle. The sample—A suspension of micro particles or blood cells—is injected into a sheath fluid streaming at a substantially higher flow rate, which assures positioning of the particles in the center of the flow channel. Particle velocities of a few m/s are achieved as required for high throughput blood cell analysis. The stability of hydrodynamic particle positioning was evaluated by measuring the pulse heights distributions of fluorescence signals from calibration beads. Quantitative assessment based on coefficient of variation for the fluorescence intensity distributions resulted in a value of about 3% determined for the micro-device exploiting cascade hydrodynamic focusing. For the spin focusing approach similar values were achieved for sample flow rates being 1.5 times lower. Our results indicate that the performances of both variants of hydrodynamic focusing suit for blood cell differentiation and counting. The potential of the micro flow cytometer is demonstrated by detecting immunologically labeled CD3 positive and CD4 positive T-lymphocytes in blood.
Keywords: microfluidics; flow cytometry; lab-on-a-chip; single cell analysis; blood cell differentiation; hydrodynamic focusing; ultraprecision milling microfluidics; flow cytometry; lab-on-a-chip; single cell analysis; blood cell differentiation; hydrodynamic focusing; ultraprecision milling
This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Frankowski, M.; Theisen, J.; Kummrow, A.; Simon, P.; Ragusch, H.; Bock, N.; Schmidt, M.; Neukammer, J. Microflow Cytometers with Integrated Hydrodynamic Focusing. Sensors 2013, 13, 4674-4693.

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