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• 電子顕微鏡による細胞内微細空間構造の解析：膜細胞骨格から核膜骨格へ
• My research works
• Bird’s-eye view of frog retina by SEM
• 視細胞外節の超微構造
• Supra molecular organization of rod outer segments
• Freeze-etched replicas of rod outer segments of photoreceptor cells
• Localization of anti-opsin antibodies：Difference in fixation
• Immuno-localization of anti-opsin
• Morphogenesis of rod outer segments
• Polarity of actin filaments that are found at the tip of the connecting cilium : S1 decoration
• Electron micrographs of synaptic ribbons
• Synaptic ribbons observed from various points of view at mesoscale
• Anti-Caveolin 1 antibodies are found on the synaptic ribbon
• Elastic and inelastic scattered electrons
• Optical pass way in the electron microscope equipped with γ type energy filter
• EELS imaging of Ca in cone synaptic pedicle
• Ca localization determined by EELS imaging
• アクチン細胞骨格の空間構造と空間特異性
• Purpose
• Membrane and cytoplasmic cytoskeleton
• Spatial distribution of membrane actin filaments
• Live cell imaging of GFP-actin in NRK cells
• Cytoplasmic surface of cell membrane covered with many actin filaments
• Clathrin and actin filaments
• Few membrane cytoskeletons are detected even in grazing thin section,
• Tomography of membrane cytoskeleton
• Actin filaments are classified into three types based on spatial distribution
• AFM imaging of membrane cytoskeleton in water
• AFM imaging of membrane cytoskeleton in water (2)
• AFM imaging of clathrin coats in water
• Whole cell observation
• Cryo-electron micrograph of native cytoskeleton beneath the cell membrane
• Cryo-electron micrograph of native cytoskeleton beneath the cell membrane (2)
• Cryo-electron micrograph of native cytoskeleton beneath the cell membrane (3)
• Spatial Specificity of Actin Cytoskeletons found by immuno-labeling
• Protocol of immuno-freeze-etching
• Actin filaments are identified by labeling with colloidal gold conjugated antibody
• IQGAP 1
• VASP
• Cdc 42
• Rac 5nm gold / Sla 1 10nm gold
• N WASP 10nm gold / IQGAP 1 5 nm gold
• Myosin II appeared to be more abundant on stress fibers than the other type of actin filaments on the membrane.
• Focal contact related proteins [Integrin b1]
• Focal contact related proteins [Zyxin]
• Immuno-gold labeling (IQCAP-1) in cryo-electron microscopy
• Cryo-electron micrograph showing the dominant area of caveoli labeled with anti-IQGAP1 immunogold
• Summary １
• Cytoplasmic and Peri-nuclear actin filaments
• CLSM visualize the cytoskeleton mainly at a bottom plane.
• Cytoplasmic cytoskeleton observed by extremely high voltage (1000KV) TEM.
• High voltage electron microscopic image of unroofed pigment epithelial cells.
• Myosin II localization in cytoplasmic cytoskeleton
• High resolution SEM image of cytoplasmic cytoskeleton
• Actin filaments in the periphery of nucleus
• Many actin filaments are extending from the nuclear envelope
• Actin filaments are associated intimately with intermediate filaments on the nuclear envelope.
• Grazing section of the nucleus labeled with SI
• Actin and intermediate filaments originating from nuclear pores (periphery)
• Back scattered electron image showing vimentin filaments extending from nuclear pores
• Anti-vimentin immuno-golds were found predominantly in periphery of the nucleus.
• Intermediate filaments covering the outer nuclear envelope were identified as Vimentin.
• Anti-Nesprin 1 antibodies are detected predominantly near nuclear pores.
• Summary ２
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