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.
A microfluidic manifold has been designed, fabricated, and tested that hydrodynamically focuses a sample into the center of a microchannel and provides control over the vertical position of the sample via the flowrates of the focusing fluids. To characterize the focusing action, a mixing experiment was performed in which the sample fluid and focusing fluid contained different fluorescent dyes. By sweeping the ratio of the rate of the top focusing fluid to the rate of the bottom focusing fluid, the sample was positioned first near the top of the microchannel and then translated downward in steps to the bottom of the microchannel. Images were obtained with confocal microscopy, and the presumptive concentration distributions were computed using multiphysics software. The simulations were shown by direct visual comparison with the experimental images to accurately predict the distributions of fluids in our device. In order to quantitatively compare the two data sets, the images and simulations were analyzed using a simple center-of-mass measurement, and according to this measurement, the simulations accurately predicted the vertical position the focused sample.
Low-power electrolysis-based microfluidic pumps utilizing the principle of hydraulics, integrated with microfluidic channels in polydimethylsiloxane (PDMS) substrates, are presented. The electro-hydraulic pumps (EHPs), consisting of electrolytic, hydraulic and fluidic chambers, were investigated using two types of electrodes: stainless steel for larger volumes and annealed gold electrodes for smaller-scale devices. Using a hydraulic fluid chamber and a thin flexible PDMS membrane, this novel prototype successfully separates the reagent fluid from the electrolytic fluid, which is particularly important for biological and chemical applications. The hydraulic advantage of the EHP device arises from the precise control of flow rate by changing the electrolytic pressure generated, independent of the volume of the reagent chamber, mimicking the function of a hydraulic press. Since the reservoirs are pre-filled with reagents and sealed prior to testing, external fluid coupling is minimized. The stainless steel electrode EHPs were manufactured with varying chamber volume ratios (1 : 1 to 1 : 3) as a proof-of-concept, and exhibited flow rates of 1.25 to 30 microl/min with electrolysis-based actuation at 2.5 to 10 V(DC). The miniaturized gold electrode EHPs were manufactured with 3 mm diameters and 1 : 1 chamber volume ratios, and produced flow rates of 1.24 to 7.00 microl/min at 2.5 to 10 V(AC), with a higher maximum sustained pressure of 343 KPa, suggesting greater device robustness using methods compatible with microfabrication. The proposed technology is low-cost, low-power and disposable, with a high level of reproducibility, allowing for ease of fabrication and integration into existing microfluidic lab-on-a-chip and analysis systems.
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