Student Assistant (Dr.Neeves) CU Denver Mead, Colorado, United States
Introduction: Platelet purification is an essential technique employed to eradicate undesirable constituents from platelet-rich plasma (PRP). This is pivotal for the isolation of platelets for analytical purposes, including the examination of platelet dispersion, metabolic activity, and their function in flow-based assays. Conventional methods of platelet purification are often marred by the inadvertent activation and clumping of platelets, which culminates in a suboptimal recovery of unadulterated and viable platelets. To address this, the present device shows the opportunity to harnesses microfluidic technology to refine the differentiation of platelets from plasma impurities by incorporating a Dean flow mechanism.
Materials and
Methods: Parameters A mask was created using a 5-loop spiral design with two inlets and two outlets using AutoCAD. Channels (W: 120 µm, H: 60 µm) were spaced 230 µm from each other. The initial radius of curvature of the spiral is 3 mm and the total length of the microchannels is ~13 cm.
Photolithography/Device Fabrication The spiral channel was fabricated on a 4’’ silicon wafer using standard photolithography. KMPR 1035 was used to achieve the desired height of 60 um. PDMS was used to generate the spiral devices containing the channels in a 10:1 polymer to cross-linker ratio. Devices were plasma bonded to glass slides treated with 12M HCl and MeOH
Characterization One syringe contained 1 µm FITC fluorescently labelled beads in 0.05% SDS. The other syringe contained DI water. Using a Harvard syringe pump, the two solutions were perfused at varying flow rates that were calculated. Images were obtained using the Olympus IX81 microscope.
Results, Conclusions, and Discussions: Preliminary findings indicate that for the isolation process involving 1.0µm diameter beads, a Dean force coefficient of 0.47 yields optimal separation results that parallel the intended outcomes for platelet purification. However, further experimentation is required to substantiate these results due to anomalies encountered in several test iterations. Given that platelets typically range from 2-3µm in diameter, it is imperative to extend the investigation to include beads that are commensurate in size to ensure the relevance and applicability of the data to platelet separation techniques.
