A Microfluidic Culture Platform to Assess Axon Degeneration

Methods Mol Biol. 2020:2143:83-96. doi: 10.1007/978-1-0716-0585-1_7.

Abstract

The field of microfluidics allows for the precise spatial manipulation of small amounts of fluids. Within microstructures, laminar flow of fluids can be exploited to control the diffusion of small molecules, creating desired microenvironments for cells. Cellular neuroscience has benefited greatly from devices designed to fluidically isolate cell bodies and axons. Microfluidic devices specialized for neuron compartmentalization are made of polydimethylsiloxane (PDMS) which is gas permeable, is compatible with fluorescence microscopy, and has low cost. These devices are commonly used to study signals initiated exclusively on axons, somatodendritic compartments, or even single synapses. We have also found that microfluidic devices allow for rapid, reproducible interrogation of axon degeneration. Here, we describe the methodology for assessing axonal degeneration in microfluidic devices. We describe several use cases, including enucleation (removal of cell bodies) and trophic deprivation to investigate axon degeneration in pathological and developmental scenarios, respectively.

Keywords: Axon degeneration; Injury; Microfluidic devices; NGF deprivation; PDMS; Wallerian degeneration.

MeSH terms

  • Animals
  • Axons / physiology*
  • Axotomy
  • Cells, Cultured
  • Dimethylpolysiloxanes
  • Equipment Design
  • Immunohistochemistry / methods
  • Intravital Microscopy / methods
  • Lab-On-A-Chip Devices*
  • Mice
  • Microfluidic Analytical Techniques / instrumentation
  • Microfluidic Analytical Techniques / methods*
  • Nerve Growth Factor / pharmacology
  • Random Allocation
  • Reproducibility of Results
  • Sensory Receptor Cells / ultrastructure
  • Single-Blind Method
  • Superior Cervical Ganglion / cytology
  • Wallerian Degeneration / physiopathology*

Substances

  • Dimethylpolysiloxanes
  • baysilon
  • Nerve Growth Factor