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4 ARTICLES PUBLISHED IN JoVE

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Biology

Long-term, High-resolution Confocal Time Lapse Imaging of Arabidopsis Cotyledon Epidermis during Germination
Kylee M. Peterson 1, Keiko U. Torii 1,2,3
1Department of Biology, University of Washington, 2Howard Hughes Medical Institute, University of Washington, 3PRESTO, Japan Science and Technology Agency

We describe a protocol using chamber slides and media to immobilize plant cotyledons for confocal imaging of the epidermis over several days of development, documenting stomatal differentiation. Fluorophore-tagged proteins can be tracked dynamically by expression and subcellular localization, increasing understanding of their possible roles during cell division and cell-type differentiation.

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Bioengineering

Photopatterning Proteins and Cells in Aqueous Environment Using TiO2 Photocatalysis
Hideaki Yamamoto 1,2, Takanori Demura 3, Kohei Sekine 3, Sho Kono 3, Michio Niwano 2,4, Ayumi Hirano-Iwata 2,5, Takashi Tanii 6
1Frontier Research Institute for Interdisciplinary Sciences, Tohoku University, 2CREST, Japan Science and Technology Agency, 3School of Fundamental Science and Engineering, Waseda University, 4Research Institute of Electrical Communication, Tohoku University, 5Graduate School of Biomedical Engineering, Tohoku University, 6Faculty of Science and Engineering, Waseda University

We describe a protocol for modifying cell affinity of a scaffold surface in aqueous environment. The method takes advantage of titanium dioxide photocatalysis to decompose organic film in the photo-irradiated region. We show that it can be used to create microdomains of scaffolding proteins, both ex situ and in situ.

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Engineering

Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets
Masamune Morita 1, Hitoyoshi Yamashita 1,2, Masayuki Hayakawa 1, Hiroaki Onoe 3, Masahiro Takinoue 1,4
1Department of Computational Intelligence and Systems Science, Interdisciplinary Graduate School of Science and Engineering, Tokyo Institute of Technology, 2Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 3Department of Mechanical Engineering, Keio University, 4PRESTO, Japan Science and Technology Agency

Here, we demonstrate a simple production method for size-controllable, monodisperse, water-in-oil (W/O) microdroplets using a capillary-based centrifugal microfluidic device. This method requires only a small sample volume and enables high-yield production. We expect this method will be useful for rapid biochemical and cellular analyses.

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Chemistry

Self-Assembly of Hybrid Lipid Membranes Doped with Hydrophobic Organic Molecules at the Water/Air Interface
Xingyao Feng 1, Teng Ma 2, Daisuke Tadaki 1, Ayumi Hirano-Iwata 1,2
1Research Institute of Electrical Communication, Tohoku University, 2Advanced Institute for Materials Research, Tohoku University

We report a protocol for producing a hybrid lipid membrane at the water/air interface by doping the lipid bilayer with copper (II) 2,9,16,23-tetra-tert-butyl-29H,31H-phthalocyanine (CuPc) molecules. The resulting hybrid lipid membrane has a lipid/CuPc/lipid sandwich structure. This protocol can also be applied to the formation of other functional nanomaterials.

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