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. 2014 Feb 19;8(1):014108.
doi: 10.1063/1.4864235. eCollection 2014 Jan.

Using microfluidic chip to form brain-derived neurotrophic factor concentration gradient for studying neuron axon guidance

Hui Huang  1 , Lili Jiang  1 , Shu Li  2 , Jun Deng  1 , Yan Li  1 , Jie Yao  1 , Biyuan Li  1 , Junsong Zheng  1
Affiliations

Using microfluidic chip to form brain-derived neurotrophic factor concentration gradient for studying neuron axon guidance

Hui Huang et al. Biomicrofluidics. .

Abstract

Molecular gradients play a significant role in regulating biological and pathological processes. Although conventional gradient-generators have been used for studying chemotaxis and axon guidance, there are still many limitations, including the inability to maintain stable tempo-spatial gradients and the lack of the cell monitoring in a real-time manner. To overcome these shortcomings, microfluidic devices have been developed. In this study, we developed a microfluidic gradient device for regulating neuron axon guidance. A microfluidic device enables the generation of Brain-derived neurotrophic factor (BDNF) gradient profiles in a temporal and spatial manner. We test the effect of the gradient profiles on axon guidance, in the BDNF concentration gradient axon towards the high concentration gradient. This microfluidic gradient device could be used as a powerful tool for cell biology research.

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Figures

Figure 1
Figure 1
Chip fabrication and concentration gradients generation. (a) Schematic of gradient microfluidic chip. (b) Divided the cell culture chamber into six regions. (c) The SU-8 mold of gradients microfluidic chip. (d) Using blue and orange dyes to make the gradient formation visualization. (e) Using fluorescein to quantitatively analyse gradient profile. (f) The relationship between fluorescent intensity and the width of cell culture chamber.
Figure 2
Figure 2
The method to evaluate the neuron guidance angle.
Figure 3
Figure 3
Neurons response in the BDNF gradient and BDNF uniform microfluidic chip. (a) Population view of the neurons in the BDNF gradient microfluidic chip. From ① to ④, the concentration of BDNF concentration is increased, cell numbers increased too. (b) The number of neurons in different regions of BDNF gradient chip. (c) The direction of axon in the gradient microfluidic chip. Most of axons towards the high concentration gradient. (d) The direction of axon in the BDNF uniform microfluidic chip.
See this image and copyright information in PMC

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