We have determined the parameters necessary to fabricate reproducible neuronal patterns which
we are using to begin studying fundamental issues in developmental neurobiology. The addition of a beam
homogenizer, as well as a new surface preparation, has enabled the routine production of reproducible, high-resolution (2−20 μm) organosilane patterns. The effects of surface preparation and beam dosage were monitored
using X-ray photoelectron spectroscopy (XPS) and proof of patterning is provided by high-resolution imaging
XPS. We also report the guidance of neuronal adhesion and neurite outgrowth and the creation of reproducibly
defined circuits of embryonic (E18−19) rat hippocampal neurons using these patterned surfaces in vitro. We
have achieved a >50% rate of pattern formation, and at times the rate approaches 90%. We are using these
patterns to address the issue of how geometric pattern cues might be used to affect cell-to-cell communication
and we report the preliminary results on the synaptic development of the hippocampal neurons using dual
patch-clamp electrophysiology. We monitored neurite outgrowth and the emergence of both spontaneous and
evoked synaptic activity for both patterned and unpatterned (control) hippocampal cultures. The results indicate
the intriguing possibility that geometry itself may be a modulating or trophic factor for cell development.
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