Microfluidic Adaptation of Density-Gradient Centrifugation for Isolation of Particles and Cells

Abstract

Density-gradient centrifugation is a label-free approach that has been extensively used for cell separations. Though elegant, this process is time-consuming (>30 min), subjects cells to high levels of stress (>350 g) and relies on user skill to enable fractionation of cells that layer as a narrow band between the density-gradient medium and platelet-rich plasma. We hypothesized that microfluidic adaptation of this technique could transform this process into a rapid fractionation approach where samples are separated in a continuous fashion while being exposed to lower levels of stress (<100 g) for shorter durations of time (<3 min). To demonstrate proof-of-concept, we designed a microfluidic density-gradient centrifugation device and constructed a setup to introduce samples and medium like Ficoll in a continuous, pump-less fashion where cells and particles can be exposed to centrifugal force and separated via different outlets. Proof-of-concept studies using binary mixtures of low-density polystyrene beads (1.02 g/cm3) and high-density silicon dioxide beads (2.2 g/cm3) with Ficoll–Paque (1.06 g/cm3) show that separation is indeed feasible with >99% separation efficiency suggesting that this approach can be further adapted for separation of cells.

Keywords

• cell separations
• label-free cell separation
• microfluidics
• density-gradient centrifugation