title: Long-Term Growth of Moss in Microfluidic Devices Enables Subcellular Studies in Development.
abstract: Key developmental processes that occur on the subcellular and cellular level or occur in occluded tissues are difficult to access, let alone image and analyze. Recently, culturing living samples within polydimethylsiloxane (PDMS) microfluidic devices has facilitated the study of hard-to-reach developmental events. Here, we show that an early diverging land plant, Physcomitrella patens, can be continuously cultured within PDMS microfluidic chambers. Because the PDMS chambers are bonded to a coverslip, it is possible to image P. patens development at high resolution over long time periods. Using PDMS chambers, we report that wild-type protonemal tissue grows at the same rate as previously reported for growth on solid medium. Using long-term imaging, we highlight key developmental events, demonstrate compatibility with high-resolution confocal microscopy, and obtain growth rates for a slow-growing mutant. By coupling the powerful genetic tools available to P. patens with long-term growth and imaging provided by PDMS microfluidic chambers, we demonstrate the capability to study cellular and subcellular developmental events in plants directly and in real time.
authors.affiliation: Department of Biology (C.S.B., S.-Z.W., M.B.) and Plant Biology Graduate Program (C.S.B.), University of Massachusetts, Amherst, Massachusetts 01003; andDepartment of Chemical Engineering (K.N., J.O.) and Department of Molecular Biology (K.N.), University of Wyoming, Laramie, Wyoming 82071.
authors.forename: Carlisle S
authors.identifier: 0000-0003-0188-8657
authors.initials: CS
authors.lastname: Bascom
authors.affiliation: Department of Biology (C.S.B., S.-Z.W., M.B.) and Plant Biology Graduate Program (C.S.B.), University of Massachusetts, Amherst, Massachusetts 01003; andDepartment of Chemical Engineering (K.N., J.O.) and Department of Molecular Biology (K.N.), University of Wyoming, Laramie, Wyoming 82071.
authors.forename: Shu-Zon
authors.identifier: 0000-0003-1387-2011
authors.initials: SZ
authors.lastname: Wu
authors.affiliation: Department of Biology (C.S.B., S.-Z.W., M.B.) and Plant Biology Graduate Program (C.S.B.), University of Massachusetts, Amherst, Massachusetts 01003; andDepartment of Chemical Engineering (K.N., J.O.) and Department of Molecular Biology (K.N.), University of Wyoming, Laramie, Wyoming 82071.
authors.forename: Katherine
authors.identifier: 0000-0002-1070-1551
authors.initials: K
authors.lastname: Nelson
authors.affiliation: Department of Biology (C.S.B., S.-Z.W., M.B.) and Plant Biology Graduate Program (C.S.B.), University of Massachusetts, Amherst, Massachusetts 01003; andDepartment of Chemical Engineering (K.N., J.O.) and Department of Molecular Biology (K.N.), University of Wyoming, Laramie, Wyoming 82071.
authors.forename: John
authors.identifier: 0000-0003-1023-1964
authors.initials: J
authors.lastname: Oakey
authors.affiliation: Department of Biology (C.S.B., S.-Z.W., M.B.) and Plant Biology Graduate Program (C.S.B.), University of Massachusetts, Amherst, Massachusetts 01003; andDepartment of Chemical Engineering (K.N., J.O.) and Department of Molecular Biology (K.N.), University of Wyoming, Laramie, Wyoming 82071 bezanilla@bio.umass.edu.
authors.forename: Magdalena
authors.identifier: 0000-0001-6124-9916
authors.initials: M
authors.lastname: Bezanilla
mesh_terms: D019068:Bryopsida / Q000166:cytology / Q000235:genetics / Q000254:growth & development*; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D056656:Lab-On-A-Chip Devices; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D018613:Microscopy, Confocal; D009154:Mutation; D015203:Reproducibility of Results; D013997:Time Factors; D059008:Time-Lapse Imaging / Q000379:methods; D046509:Tissue Culture Techniques / Q000295:instrumentation / Q000379:methods
chemical_list: D004129:Dimethylpolysiloxanes; C013830:baysilon