title: Parylene mobile microplates integrated with an enzymatic release for handling of single adherent cells.
abstract: An approach for manipulating single adherent cells is developed that is integrated with an enzymatic batch release. This strategy uses an array of releasable microfabricated mobile substrates, termed microplates, formed from a biocompatible polymer, parylene. A parylene microplate array of 10-70 μm in diameter can be formed on an alginate hydrogel sacrificial layer by using a standard photolithographic process. The parylene surfaces are modified with fibronectin to enhance cell attachment, growth, and stretching. To load single cells onto these microplates, cells are initially placed in suspension at an optimized seeding density and are allowed to settle, stretch, and grow on individual microplates. The sacrificial layer underneath the microplate array can be dissolved on a time-scale of several seconds without cytotoxicity. This system allows the inspection of selected single adherent cells. The ability to assess single cells while maintaining their adhesive properties will broaden the examination of a variety of attributes, such as cell shape and cytoskeletal properties.
authors.affiliation: Institute of Industrial Science, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8505, Japan.
authors.forename: Tetsuhiko
authors.initials: T
authors.lastname: Teshima
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Onoe
authors.forename: Kaori
authors.initials: K
authors.lastname: Kuribayashi-Shigetomi
authors.forename: Hiroka
authors.initials: H
authors.lastname: Aonuma
authors.forename: Koki
authors.initials: K
authors.lastname: Kamiya
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Ishihara
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Kanuka
authors.forename: Shoji
authors.initials: S
authors.lastname: Takeuchi
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D004798:Enzymes / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology* / Q000469:parasitology; D006801:Humans; D008297:Male; D051379:Mice; D011108:Polymers / Q000737:chemistry*; D051381:Rats; D059010:Single-Cell Analysis / Q000295:instrumentation*; D014122:Toxoplasma / Q000502:physiology; D014992:Xylenes / Q000737:chemistry*
chemical_list: D004798:Enzymes; D011108:Polymers; D014992:Xylenes; C011055:parylene
keywords: enzymatic release; microfluidic devices; mobile microplates; parasitic infections; single-cell analysis