Hydrodynamic optical alignment for microflow cytometry

Abstract

A microfabricated flow cytometer has been developed that is capable of detecting nearly all of the microparticles in an aqueous suspension. Current design allows for integrated coupling between an optical fiber-based detection system and the particle stream via hydrodynamic focusing. By adjusting the relative flow-rates at the auxiliary inputs of the focusing manifold, the particle stream can be steered out-of-plane relative to the illuminating laser, and similarly the particle stream can be squeezed or expanded. The microfabricated device was constructed in polydimethylsiloxane with cross-sectional microfluidic dimensions of 125 µm × 125 µm. Using the present device and method, fluorescent microparticles in aqueous solution were counted at an absolute counting efficiency of 91 ± 4%. The coefficient of variation of the fluorescence pulse-heights for far-red fluorescent microparticles was 15%. The device exhibited a linear response to fluorescence intensity calibration microparticles as shown by comparison with a commercial cytometer instrument.