Miniaturized flow cytometer with 3D hydrodynamic particle focusing and integrated optical elements applying silicon photodiodes

In this study, the design, realization and measurement results of a novel optofluidic system capable of performing absorbance-based flow cytometric analysis is presented. This miniaturized laboratory platform, fabricated using SU-8 on a silicon substrate, comprises integrated polymer-based waveguides for light guiding and a biconcave cylindrical lens for incident light focusing. The optical structures are detached from the microfluidic sample channel resulting in a significant increase in optical sensitivity. This allows the application of standard solid-state laser and standard silicon-based photodiodes operated by lock-in-amplification resulting in a highly practical and effective detection system. The easy-to-fabricate single-layer microfluidic structure enables independently adjustable 3D hydrodynamic sample focusing to an arbitrary position in the channel. To confirm the fluid dynamics and raytracing simulations and to characterize the system, different sets of microparticles and T-lymphocyte cells (Jurkat cell line) for vital staining were investigated by detecting the extinction (axial light loss) signal. The analytical classification via signal peak height/width demonstrates the high sensitivity and sample discrimination capability of this compact low-cost/low-power microflow cytometer.