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Molecular Surveillance of Viral Processes Using Silicon Nitride Membranes
Virginia Tech Carilion Research Institute, 2 Riverside Circle, Roanoke, VA 24016, USA
Protochips, Inc., 616 Hutton Street, Raleigh, NC 27606, USA
These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 5 November 2012; in revised form: 18 December 2012 / Accepted: 5 March 2013 / Published: 14 March 2013
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
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MDPI and ACS Style
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.
Gilmore BL, Tanner JR, McKell AO, Boudreaux CE, Dukes MJ, McDonald SM, Kelly DF. Molecular Surveillance of Viral Processes Using Silicon Nitride Membranes. Micromachines. 2013; 4(1):90-102.
Gilmore, Brian L.; Tanner, Justin R.; McKell, Allison O.; Boudreaux, Crystal E.; Dukes, Madeline J.; McDonald, Sarah M.; Kelly, Deborah F. 2013. "Molecular Surveillance of Viral Processes Using Silicon Nitride Membranes." Micromachines 4, no. 1: 90-102.