UBC Theses and Dissertations
Multiplexed end-point microfluidic chemotaxis assay using centrifugal alignment Satti, Ramaa Krishna Sampath
A fundamental challenge to the scalability of microfluidic chemotaxis assays is the requirement for time-lapse imaging to continuously track migrating cells. Current assays are not suitable for drug testing and drug screening applications that require the ability to perform hundreds of experiments in parallel. End-point chemotaxis assays wherein cells are aligned at fixed starting point before migration have been proposed as an alternative to continuous tracking. Previous methods developed to align cells use fluid flow to pattern cells in traps or constrictions, requiring external instrumentation and subjecting the cells to high shear stress. By patterning cells through centrifugation in our microfluidic device, alignment can be achieved without precise flow control while applying minimal shear stress to cells. This technique is insensitive to cell geometry and is capable of handling rare cells in the liquid sample. Additionally, as the chemical gradient in the assay is generated through passive diffusion, the stand-alone device can be placed in the incubator and subsequently imaged to obtain the migration characteristics in each of the 12 devices on our substrate. The device was used to observe the response of human neutrophils to gradients of fMLP. Our study reveals the potential to leverage cell alignment through centrifugation to develop highly scalable, end-point microfluidic chemotaxis assays.
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