HIGH-THROUGHPUT COMBINATORIAL STUDY OF AXON INITIATION AND GUIDANCE OF SINGLE NEURONS IN RESPONSE TO MICROFLUIDIC BIOCHEMICAL GRADIENTS

摘要

In this paper, we report the development of a high-throughput microfluidic gradient generator array for the study of axon growth and guidance dynamics of 512 individual primary mammalian neurons in parallel. We adapt a previously reported microfluidic method of creating gradients on open-surfaces termed "microjets" [1], combine it with live-cell, auto-focus time-lapse imaging to create an integrated platform. This format (a) gives us statistically-rich data from a single experiment, (b) produces a stable gradient with negligible shear forces on the culture surface, (c) is amenable to long-term culture of primary mammalian neurons, and (d) allows for the presentation of different concentration gradients of two opposing biochemical molecules.