title: A self-reconfiguring metamorphic nanoinjector for injection into mouse zygotes.
abstract: This paper presents a surface-micromachined microelectromechanical system nanoinjector designed to inject DNA into mouse zygotes which are ≈90 μm in diameter. The proposed injection method requires that an electrically charged, DNA coated lance be inserted into the mouse zygote. The nanoinjector's principal design requirements are (1) it must penetrate the lance into the mouse zygote without tearing the cell membranes and (2) maintain electrical connectivity between the lance and a stationary bond pad. These requirements are satisfied through a two-phase, self-reconfiguring metamorphic mechanism. In the first motion subphase a change-point six-bar mechanism elevates the lance to ≈45 μm above the substrate. In the second motion subphase, a compliant folded-beam suspension allows the lance to translate in-plane at a constant height as it penetrates the cell membranes. The viability of embryos following nanoinjection is presented as a metric for quantifying how well the nanoinjector mechanism fulfills its design requirements of penetrating the zygote without causing membrane damage. Viability studies of nearly 3000 nanoinjections resulted in 71.9% of nanoinjected zygotes progressing to the two-cell stage compared to 79.6% of untreated embryos.
authors.affiliation: Nexus Spine, LLC, Salt Lake City, Utah 84124, USA.
authors.forename: Quentin T
authors.initials: QT
authors.lastname: Aten
authors.affiliation: Department of Mechanical Engineering, Brigham Young University, Provo, Utah 84602, USA.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Jensen
authors.affiliation: Department of Microbiology and Molecular Biology, Brigham Young University, Provo, Utah 84602, USA.
authors.forename: Sandra H
authors.initials: SH
authors.lastname: Burnett
authors.affiliation: Department of Mechanical Engineering, Brigham Young University, Provo, Utah 84602, USA.
authors.forename: Larry L
authors.initials: LL
authors.lastname: Howell
mesh_terms: D000818:Animals; D004247:DNA / Q000494:pharmacology*; D018014:Gene Transfer Techniques / Q000295:instrumentation*; D051379:Mice; D008845:Microinjections / Q000295:instrumentation / Q000379:methods; D015053:Zygote / Q000166:cytology*
chemical_list: D004247:DNA