-lab. 研究概要 細胞の生命活動konishi/mems/gbc/ja-gbc.pdf-lab. © konishi-lab. ©...
TRANSCRIPT
© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L
© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L
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研究概要 1/4
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細胞の生命活動細胞の生命活動
神経細胞
イオンイオン
電気信号ピペット
検出電極
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© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L心筋細胞
イオンイオン
<単一細胞><細胞群> パッチクランプ法
細胞観測観測
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Mi Ch l A (MCA)
課題点課題点 ・ ロースループット・ ピペット製作、操作が困難
MEMS技術
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細胞吸引孔 と 電極 を一体化 した 構造
Micro Channel Array (MCA)絶縁膜開口部:
Φ10 μm
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© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L5 mm
吸引固定 による 細胞選択性 と S/N比の向上
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Micro / Nano Mechatronics LAB.Micro / Nano Mechatronics LAB. KonishiKonishi LAB.LAB.Group CellGroup Cell
5 mm
吸引孔:Φ4 μm 所望の組織形成・定量解析 に向けた 基本構造
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研究目的 2/4
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細胞外解析細胞外解析細胞外解析細胞外解析
MCA を基本構造とした 細胞操作細胞操作 ・・ 定量解析定量解析 ツール の 開発
細胞 解析細胞 解析細胞 解析細胞 解析© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L
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細胞外解析用ツール細胞外解析用ツール細胞外解析用ツール細胞外解析用ツール 細胞内解析用ツール細胞内解析用ツール細胞内解析用ツール細胞内解析用ツール
電極針 バネ支持梁細胞 絶縁膜
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© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L
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© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L支援ポール 吸引孔
ラッチ
ガイド構造陰圧
電極
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© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L
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細胞内挿入用針電極型細胞内挿入用針電極型 MCAMCA細胞 溶液溜
細胞ネットワーク解析用細胞ネットワーク解析用 MCAMCA陰圧
細胞(磁性粒子)
空気圧
磁路一体化微小孔
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© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L電極ON OFF電磁石
冶具
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Micro / Nano Mechatronics LAB.Micro / Nano Mechatronics LAB. KonishiKonishi LAB.LAB.Group CellGroup Cell
空気圧空気圧 ++ 磁気誘導磁気誘導 MCAMCA 化学的細胞膜穿孔用化学的細胞膜穿孔用 MCAMCA
電極ON OFF電磁石
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研究成果 3/4
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細胞外解析用ツール細胞外解析用ツール細胞外解析用ツール細胞外解析用ツール 細胞内解析用ツール細胞内解析用ツール細胞内解析用ツール細胞内解析用ツール
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駆動源(プローブ) 吸引孔
ガイド構造
1 sec 1 sec
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電極針
ラッチ
1 sec 1 sec 細胞内挿入用針電極型 MCA の駆動の様子© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L
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スライス自発活動電位の多点同時観測結果
hv細胞培地
1. 培地充填 2. 細胞吸引 3. 溶液交換 4. 細胞膜穿孔
OFF
磁性細胞(Φ10-20 μm)
流れ (40 nl / s)
OFF
(磁気)流れ (40 nl / s)
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光増感剤+ Lucifer Yellow 電極陰圧
吸引孔吸引孔 細胞細胞((PC12PC12)) LYLY染色された染色された細胞細胞
(Φ10 20 μm)
200 μm
磁路一体化
OFF
ON
OFF
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Micro / Nano Mechatronics LAB.Micro / Nano Mechatronics LAB. KonishiKonishi LAB.LAB.Group CellGroup Cell
単一細胞の磁気操作
30 μm
化学的細胞膜穿孔の実験結果
ON磁路 体化微小孔
磁気誘導@ 70 mA
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まとめ 4/4磁路一体化
微小孔© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L
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細胞操作・解析用ツール細胞操作・解析用ツール のの 開発開発
細胞外10
パーマロイ
Φ20 μm
200 μm
微小孔
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細胞ネットワーク解析用MCA
空気圧+磁気誘導MCA
細胞内
細胞ネットワーク解析用細胞ネットワーク解析用 MCAMCA空気圧空気圧 ++ 磁気誘導磁気誘導 MCAMCA
10 mm
染色された細胞
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細胞内
細胞内挿入用針電極型細胞内挿入用針電極型 MCAMCA化学的細胞膜穿孔用化学的細胞膜穿孔用 MCAMCA
Suction hole
Needle probe
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© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L関連発表
細胞内挿入用針電極型MCA
細胞・細胞ネットワーク定量解析への応用細胞・細胞ネットワーク定量解析への応用hole 30 μm
化学的細胞膜穿孔用MCA
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関連発表
○T.Hiranishi, W.Tonomura, K.Ino, M.Okochi, H.Honda and S.Konishi, “Both Pneumatic and Magnetic Induction of Scattered Cells on Micro Channel Arrayfor Cellular Analysis”, IEEE International Conference on Micro Electro Mechanical Systems (MEMS2008), pp.284-287, Tucson, Arizona, USA, 2008.
○W.Tonomura, T.Kurashima, Y.Takayama, H.Moriguchi, Y.Jimbo and S.Konishi, “Simultaneous Multipoint Measurement of Cellular Network by IsolatedMicro Channel Array with Pt-Black Electrodes”, IEEE International Conference on Transducers & Eurosensors, pp.1793-1796, Lyon, France, 2007. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L© Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab. © Konishi-Lab.-Lab. © Konishi-L
Micro / Nano Mechatronics LAB.Micro / Nano Mechatronics LAB. KonishiKonishi LAB.LAB.Group CellGroup Cell
y pp y
○K.Iso, T.K.Saito, H.Muguruma, H.Tabata and S.Konishi, “Photodynamic Perforation of Cell Membrane on Micro Channel Array toward Intracellular Technology”, IEEE International Conference on Micro Electro Mechanical Systems (MEMS2007), pp.445-448, Kobe, Japan, 2007.