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Micromachines 2013, 4(1), 90-102; doi:10.3390/mi4010090
Article

Molecular Surveillance of Viral Processes Using Silicon Nitride Membranes

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Received: 5 November 2012 / Revised: 18 December 2012 / Accepted: 5 March 2013 / Published: 14 March 2013
(This article belongs to the Special Issue Micromachined Tools for Nanoscale Science and Technology)
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Abstract

Here we present new applications for silicon nitride (SiN) membranes to evaluate biological processes. We determined that 50-nanometer thin films of SiN produced from silicon wafers were sufficiently durable to bind active rotavirus assemblies. A direct comparison of SiN microchips with conventional carbon support films indicated that SiN performs equivalent to the traditional substrate to prepare samples for Electron Microscopy (EM) imaging. Likewise, SiN films coated with Ni-NTA affinity layers concentrated rotavirus particles similarly to affinity-coated carbon films. However, affinity-coated SiN membranes outperformed glow-discharged conventional carbon films 5-fold as indicated by the number of viral particles quantified in EM images. In addition, we were able to recapitulate viral uncoating and transcription mechanisms directed onto the microchip surfaces. EM images of these processes revealed the production of RNA transcripts emerging from active rotavirus complexes. These results were confirmed by the functional incorporation of radiolabeled nucleotides into the nascent RNA transcripts. Collectively, we demonstrate new uses for SiN membranes to perform molecular surveillance on life processes in real-time.
Keywords: silicon nitride; microchips; rotavirus; RNA; electron microscopy silicon nitride; microchips; rotavirus; RNA; electron microscopy
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Gilmore, B.L.; Tanner, J.R.; McKell, A.O.; Boudreaux, C.E.; Dukes, M.J.; McDonald, S.M.; Kelly, D.F. Molecular Surveillance of Viral Processes Using Silicon Nitride Membranes. Micromachines 2013, 4, 90-102.

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