ベント光ファイバーを用いた共焦点蛍光顕微鏡によるミトコンドリアの力学応答

メンバー: 梅田倫弘、岩見健太郎、長崎秀昭、太田善浩
分野: 応用物理学、ナノ・マイクロ科学、基礎生物学
所属: 工学研究院
キーワード: ミトコンドリア、ヤング率、ミトコンドリア活性、屈曲光ファイバー、共焦点蛍光顕微鏡
ウェブサイト:
研究概要

心臓血管系の心筋細胞は、血流や心臓拍動によって発生する機械的応力に常にさらされている。細胞への力の効果は、様々な研究が報告されているが、ミトコンドリアのような細胞小器官への影響は未だ未解明である。そこで、我々は、先端が平坦なベント光ファイバー(BOF)と共焦点蛍光顕微鏡を用いて、ミトコンドリアにナノニュートンの外力印加を試みた。BOFチップを用いて単一ミトコンドリアに力を印加することによってミトコンドリアのヤング率は、9.6~115kPaの領域にあることが分かった。さらに、ミトコンドリアを膜電位色素で染色し、蛍光強度の変化を計測した。その結果、より活性度の高いミトコンドリアは、より高い脱分極周波数を示すことが明らかとなった。同様な傾向は、50nN以下の外力印加で観測された。さらに、100nNより大きな外力では、脱分極周波数は低下することが示された。
以上の研究結果より、ミトコンドリアの機能は、ナノニュートンレベルの外力などの物理的環境下で影響を受けることが明らかとなった。
主要論文・参考事項
1)Yongbo Li, Satoshi Hondaa, Kentaro Iwamia, Yoshihiro Ohtab, and Norihiro Umeda:Analysis of mitochondrial mechanical dynamics using a confocal fluorescence microscope with a bent optical fiber, J. of Microscopy(2015) in press
2)Takashi Asaka1, Tomohiro Itayama, Hideaki Nagasaki, Kentaro Iwami, Chizuko Yamamoto, Yukiko Hara, Atsushi Masuda, and Norihiro Umeda:Development of a pH sensor based on a nanostructured filter adding pH-sensitive fluorescent dye for detecting acetic acid in photovoltaic modules, Japanese Journal of Applied Physics (2015) in press
3)M. Ishii, K. Iwami, and N. Umeda, Appl. Phys. Lett.,106, 021115 (2015)
4)Takashi Asaka, Kentaro Iwami, Atsushi Taguchi, Norihiro Umeda, and Atsushi Masuda, Detection of acid moisture in photovoltaic modules using a dual wavelength pH-sensitive fluorescent dye, Japanese Journal of Applied Physics, 53, 04ER-1-04ER4(2014)
5)Daisuke Morikawa, Keita Kanematsu, Takahiro Shibata, Keisuke Haseda, Norihiro Umeda, and Yoshihiro Ohta, Detection of swelling of single isolated mitochondrion with optical microscopy, BIOMEDICAL OPTICS EXPRESS, 5, 3, 848-857 (2014)
6)Kentaro Iwamia, Shin Kaneko, Ryuzo Shinta, Jun Fujihara, Hideaki Nagasaki,Yasufumi Matsumura, Norihiro Umeda, Plasmon-resonance dew condensation sensor made of gold-ceramic nanocomposite and its application in condensation prevention, Sensors and Actuators B: Chemical, 184, 301-305 (2013)
7)Kentaro Iwami, Miho Ishii, Yuzuru Kuramochi, Kenichi Ida, and Norihiro Umeda, Ultrasmall radial polarizer array based on patterned plasmonic nanoslits, Appl. Phys. Lett., 101/16, 161119-1~161119-4(2012)
8)Yasuaki Kanazashi, Yongbo Li, Takumi Onojima, Kentaro Iwami, Yoshihiro Ohta, Norihiro Umeda, pH Measurement using Dual-Wavelength Fluorescent Ratio by Two-Photon Excitation for Mitochondrial Activity,Japanese Journal of Applied Physics,51/117001, 117001-1~117001-5(2012)
8)Takeshi Saitou, Muhammad Nurdin Bin, Hiroyuki Kowa, Norihiro Umeda, Kuniharu Takizawa, Eiichi Kondoh, and Lianhua Jin, Birefringence Polarimeter Using Dual LiNbO3 Electrooptic Crystal Modulators, Japanese Journal of Applied Physics, 51,82201, 082201-1~082201-5(2012)
9)Priyank Mohan, Ryuzo Shinta, Jun Fujiwara, Hiroaki Takahashi, Derrick Mott, Yasufumi Matsumura, Goro Mizutani, Kentaro Iwami, Norihiro Umeda, Shinya Maenosono, Boehmite nanorod/gold nanoparticle nanocomposite film for an easy-to-use optical humidity sensor, Sensors and Actuators B: Chemical, 168, 429-435(2012)
10)Hideki Nagasaki, Sin Kaneko, Kentaro Iwami, Norihiro Umeda, Localized Surface Plasmon Resonance Dew Sensor for Use Under Low Humidity Conditions, Japanese Journal of Applied Physics, 51, 027301-1~027301-4(2012)
11)Yongbo Li, Ryosuke Shinohara, Kentaro Iwami, Yoshihiro Ohta and Norihiro Umeda, Observation of Mitochondrial Activity Based on Temporal and Spatial pH Variation Measured by Near-Field Fluorescent Ratiometrym Science China Physics, Mechanics and Astronomy, 54, 12, 2225-2229(2011)
お問い合わせ先
東京農工大学・先端産学連携研究推進センター
urac[at]ml.tuat.ac.jp([at]を@に変換してください)
Analysis of mitochondrial mechanical dynamics using a confocal fluorescence microscope with a bent optical fiber

Research members: Dr. Norihiro Umeda, Dr. Kentarou Iwami, Dr. Hideaki Nagasaki, Dr. Yoshihiro Ohta
Research fields: Applied physics, Nano/Micro science, Basic biology
Departments: Institute of Engineering
Keywords: mitochondria, elastic moduli, mitochondrial activity, flat-ended tip, bent optical fiber, confocal fluorescence microscope
Web site:
Summary

The cells in the cardiovascular system are constantly subjected to mechanical forces created by blood flow and the beating heart. The effect of forces on cells has been extensively investigated, but their effect on cellular organelles such as mitochondria remains unclear. We examined the impact of nano-Newton forces on mitochondria using a bent optical fiber (BOF) with a flat-ended tip (diameter exceeding 2 μm) and a confocal fluorescence microscope. By indenting a single mitochondrion with the BOF tip, we found that the mitochondrial elastic modulus was proportional to the (−1/2) power of the mitochondrial radius in the 9.6–115 kPa range. We stained the mitochondria with a potential-metric dye (TMRE) and measured the changes in TMRE fluorescence intensity. We confirmed that more active mitochondria exhibit a higher frequency of repetitive transient depolarization. The same trend was observed at forces lower than 50 nN. We further showed that the depolarization frequency of mitochondria decreases under an extremely large force (nearly 100 nN). We conclude that mitochondrial function is affected by physical environmental factors, such as external forces at the nano-Newton level.
Reference articles and patents
1)Yongbo Li, Satoshi Hondaa, Kentaro Iwamia, Yoshihiro Ohtab, and Norihiro Umeda:Analysis of mitochondrial mechanical dynamics using a confocal fluorescence microscope with a bent optical fiber, J. of Microscopy(2015) in press
2)Takashi Asaka1, Tomohiro Itayama, Hideaki Nagasaki, Kentaro Iwami, Chizuko Yamamoto, Yukiko Hara, Atsushi Masuda, and Norihiro Umeda:Development of a pH sensor based on a nanostructured filter adding pH-sensitive fluorescent dye for detecting acetic acid in photovoltaic modules, Japanese Journal of Applied Physics (2015) in press
3)M. Ishii, K. Iwami, and N. Umeda, Appl. Phys. Lett.,106, 021115 (2015)
4)Takashi Asaka, Kentaro Iwami, Atsushi Taguchi, Norihiro Umeda, and Atsushi Masuda, Detection of acid moisture in photovoltaic modules using a dual wavelength pH-sensitive fluorescent dye, Japanese Journal of Applied Physics, 53, 04ER-1-04ER4(2014)
5)Daisuke Morikawa, Keita Kanematsu, Takahiro Shibata, Keisuke Haseda, Norihiro Umeda, and Yoshihiro Ohta, Detection of swelling of single isolated mitochondrion with optical microscopy, BIOMEDICAL OPTICS EXPRESS, 5, 3, 848-857 (2014)
6)Kentaro Iwamia, Shin Kaneko, Ryuzo Shinta, Jun Fujihara, Hideaki Nagasaki,Yasufumi Matsumura, Norihiro Umeda, Plasmon-resonance dew condensation sensor made of gold-ceramic nanocomposite and its application in condensation prevention, Sensors and Actuators B: Chemical, 184, 301-305 (2013)
7)Kentaro Iwami, Miho Ishii, Yuzuru Kuramochi, Kenichi Ida, and Norihiro Umeda, Ultrasmall radial polarizer array based on patterned plasmonic nanoslits, Appl. Phys. Lett., 101/16, 161119-1~161119-4(2012)
8)Yasuaki Kanazashi, Yongbo Li, Takumi Onojima, Kentaro Iwami, Yoshihiro Ohta, Norihiro Umeda, pH Measurement using Dual-Wavelength Fluorescent Ratio by Two-Photon Excitation for Mitochondrial Activity,Japanese Journal of Applied Physics,51/117001, 117001-1~117001-5(2012)
8)Takeshi Saitou, Muhammad Nurdin Bin, Hiroyuki Kowa, Norihiro Umeda, Kuniharu Takizawa, Eiichi Kondoh, and Lianhua Jin, Birefringence Polarimeter Using Dual LiNbO3 Electrooptic Crystal Modulators, Japanese Journal of Applied Physics, 51,82201, 082201-1~082201-5(2012)
9)Priyank Mohan, Ryuzo Shinta, Jun Fujiwara, Hiroaki Takahashi, Derrick Mott, Yasufumi Matsumura, Goro Mizutani, Kentaro Iwami, Norihiro Umeda, Shinya Maenosono, Boehmite nanorod/gold nanoparticle nanocomposite film for an easy-to-use optical humidity sensor, Sensors and Actuators B: Chemical, 168, 429-435(2012)
10)Hideki Nagasaki, Sin Kaneko, Kentaro Iwami, Norihiro Umeda, Localized Surface Plasmon Resonance Dew Sensor for Use Under Low Humidity Conditions, Japanese Journal of Applied Physics, 51, 027301-1~027301-4(2012)
11)Yongbo Li, Ryosuke Shinohara, Kentaro Iwami, Yoshihiro Ohta and Norihiro Umeda, Observation of Mitochondrial Activity Based on Temporal and Spatial pH Variation Measured by Near-Field Fluorescent Ratiometrym Science China Physics, Mechanics and Astronomy, 54, 12, 2225-2229(2011)
Contact
University Research Administration Center(URAC),
Tokyo University of Agriculture andTechnology
urac[at]ml.tuat.ac.jp
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