Issue 7, 2009
From the journal:

Lab on a Chip

Microfluidic structures for flow cytometric analysis of hydrodynamically focussed blood cells fabricated by ultraprecision micromachining

A. Kummrow,*a   J. Theisen,b   M. Frankowski,a   A. Tuchscheerer,b   H. Yildirim,a   K. Brattke,a   M. Schmidtb  and  J. Neukammera  

* Corresponding authors

a Physikalisch-Technische Bundesanstalt, Abbestr. 2–12, Berlin, Germany
E-mail: andreas.kummrow@ptb.de
Fax: +49-30-3481-7505
Tel: +49-30-3481-7643

b Institut für Konstruktion, Mikro- und Medizintechnik, Technische Universität Berlin, EW 1-2, Hardenbergstraße 36, Berlin, Germany

Abstract

We present three-dimensional microfluidic structures with integrated optical fibers, mirrors and electrodes for flow cytometric analysis of blood cells. Ultraprecision milling technique was used to fabricate different flow cells featuring single-stage and two-stage cascaded hydrodynamic focusing of particles by a sheath flow. Two dimensional focussing of the sample fluid was proven by fluorescence imaging in horizontal and vertical directions and found to agree satisfactorily with finite element calculations. Focussing of the sample stream down to 5 µm at a particle velocity of 3 m s−1 is accessible while maintaining stable operation for sample flow rates of up to 20 µL min−1. In addition to fluorescence imaging, the micro-flow cells were characterised by measurements of pulse shapes and pulse height distributions of monodisperse microspheres. We demonstrated practical use of the microstructures for cell differentiation employing light scatter to distinguish platelets and red blood cells. Furthermore, T-helper lymphocytes labelled by monoclonal antibodies were identified by measuring side scatter and fluorescence.

Graphical abstract: Microfluidic structures for flow cytometric analysis of hydrodynamically focussed blood cells fabricated by ultraprecision micromachining

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Article information

DOI
https://doi.org/10.1039/B808336C
Article type
Paper
Submitted
16 May 2008
Accepted
11 Dec 2008
First published
05 Jan 2009

Lab Chip, 2009,9, 972-981

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Microfluidic structures for flow cytometric analysis of hydrodynamically focussed blood cells fabricated by ultraprecision micromachining

A. Kummrow, J. Theisen, M. Frankowski, A. Tuchscheerer, H. Yildirim, K. Brattke, M. Schmidt and J. Neukammer, Lab Chip, 2009, 9, 972 DOI: 10.1039/B808336C

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