Micro flow cytometer with 3D hydrodynamic focusing

This paper reports a micro flow cytometer fabricated in Polymethylmethacrylate (PMMA) in which a 3d hydrodynamic flow focusing is employed in order to align the particles in a single line along the focused stream. The device has been fabricated by direct micro milling of two parts of PMMA that were finally bonded together. With a suitable choice of the fluidic channel geometry, a circular sample stream located in the center of the channel is obtained. Numerical simulations have been performed in order to investigate the flow characteristic of the structure and find the desiderated geometry. Three dimensional hydrofocusing of the sample fluid was analysed and demonstrated by cross sectional fluorescence imaging in good agreement with numerical simulations. Flow cytometry measurements have been performed by using 10 mu m particles. From the analysis of the fluorescence signals collected at each transit event we can confirm that the device was capable of creating a single-file particle stream. The results show that the device was capable of discriminating single microparticles with a good signal-to-noise ratio and a high throughput.