細胞および組織の研究向けAFM

AFMは細胞生物学の研究にとって非常に重要なツールです。AFMは生きた非固定の細胞の3D形状データを提供できます。しかし、細胞生物学におけるAFMの最大の強みは、正確で定量的なメカニカル測定を生理的条件に近い状態（培地中37℃）で提供できる機能にあります。細胞や基質の弾性および粘弾性応答は、フォースマップやAFMベースのマイクロレオロジーのテクニックでそれぞれごく普通に測定できます。測定される細胞のモジュラスは、変化のない細胞、異なる発達段階の細胞、分化、病気、薬剤や機械的ストレスといった刺激に対する細胞の応答になる可能性があります。細胞外基質（ECM；extracellular matrix）が細胞分化、運命、シグナル伝達、遺伝子転写、がん、心血管疾患、アポトーシスといった中で演じる役割のために、基質や細胞の微小環境のモジュラスの測定もまた重要です。
 
倒立型光学顕微鏡（蛍光、共焦点、TIRFなど）と組み合わせたとき、両方のイメージング法からのデータは、蛍光標識した構造と関連付けるために、AFM形状像と結合できます。そのオプティクスはAFMの探針に細胞の特定領域をプローブするように指示するために使用でき、イメージングが難しい細胞タイプにとって重要になります。最後に、AFMは機械的な刺激を細胞に与えるためにも使用され、その関連応答（例えば、イオンハンドリングや膜電位変化など）は、生きた細胞や組織における機械的シグナル伝達を理解するために、光学的に記録できます。
 

機能

  • 培養中の生きた細胞をイメージング
  • 細胞や基質の弾性や粘弾性応答を測定
  • AFMを倒立型光学顕微鏡や蛍光テクニックと融合
  • 光学イメージを、AFMイメージやフォース測定のための関心領域（ROI；region of interest）を選択するために使用
  • AFM形状像やモジュラスマップを、光学イメージや3D AFMイメージ上に重ね合わせ
     

一般的なアプリケーション

  • 処理後の生きた細胞の動的イメージング
  • がん細胞における剛性や粘弾性変化
  • 細胞分化における細胞基質の影響
  • 機械的な刺激に対する細胞の応答

Selected Publications

I. Acerbi, L. Cassereau, I. Dean, Q. Shi, A. Au, C. Park, Y. Y. Chen, J. Liphardt, E. S. Hwang, and V. M. Weaver, "Human breast cancer invasion and aggression correlates with ECM stiffening and immune cell infiltration," Integr. Biol. 7, 1120-1134 (2015). doi:10.1039/c5ib00040h

D. B. Agus, J. F. Alexander, W. Arap, S. Ashili, J. E. Aslan, R. H. Austin, V. Backman, K. J. Bethel, R. Bonneau et al., "A physical sciences network characterization of non-tumorigenic and metastatic cells," Sci. Rep. 3, 1449 (2013). doi:10.1038/srep01449

Z. Bálint, I. A. Krizbai, I. Wilhelm, A. E. Farkas, Á. Párducz, Z. Szegletes, and G. Váró, "Changes induced by hyperosmotic mannitol in cerebral endothelial cells: an atomic force microscopic study," Eur. Biophys. J. 36, 113-120 (2006). doi:10.1007/s00249-006-0112-4

S. Bagchi, H. Tomenius, L. M. Belova, and N. Ausmees, "Intermediate filament-like proteins in bacteria and a cytoskeletal function in Streptomyces," Mol. Microbiol. 70, 1037-1050 (2008). doi:10.1111/j.1365-2958.2008.06473.x

K. B. Bernick, T. P. Prevost, S. Suresh, and S. Socrate, "Biomechanics of single cortical neurons," Acta Biomater. 7, 1210-1219 (2011). doi:10.1016/j.actbio.2010.10.018

Z. Deng, T. Zink, H. yuan Chen, D. Walters, F. tong Liu, and G. yu Liu, "Impact of Actin Rearrangement and Degranulation on the Membrane Structure of Primary Mast Cells: A Combined Atomic Force and Laser Scanning Confocal Microscopy Investigation," Biophys. J. 96, 1629-1639 (2009). doi:10.1016/j.bpj.2008.11.015

P. C. D. P. Dingal, A. M. Bradshaw, S. Cho, M. Raab, A. Buxboim, J. Swift, and D. E. Discher, "Fractal heterogeneity in minimal matrix models of scars modulates stiff-niche stem-cell responses via nuclear exit of a mechanorepressor," Nat. Mater. 14, 951-960 (2015). doi:10.1038/nmat4350

A. J. Engler, "Myotubes differentiate optimally on substrates with tissue-like stiffness: pathological implications for soft or stiff microenvironments," J. Cell Biol. 166, 877-887 (2004). doi:10.1083/jcb.200405004

A. J. Engler, S. Sen, H. L. Sweeney, and D. E. Discher, "Matrix Elasticity Directs Stem Cell Lineage Specification," Cell 126, 677-689 (2006). doi:10.1016/j.cell.2006.06.044

N. A. Geisse, S. P. Sheehy, and K. K. Parker, "Control of myocyte remodeling in vitro with engineered substrates," In Vitro Cellular & Developmental Biology - Animal 45, 343-350 (2009). doi:10.1007/s11626-009-9182-9

J. Gingras, R. M. Rioux, D. Cuvelier, N. A. Geisse, J. W. Lichtman, G. M. Whitesides, L. Mahadevan, and J. R. Sanes, "Controlling the Orientation and Synaptic Differentiation of Myotubes with Micropatterned Substrates," Biophys. J. 97, 2771-2779 (2009). doi:10.1016/j.bpj.2009.08.038

F. M. Hecht, J. Rheinlaender, N. Schierbaum, W. H. Goldmann, B. Fabry, and T. E. Schäffer, "Imaging viscoelastic properties of live cells by AFM: power-law rheology on the nanoscale," Soft Matter 11, 4584-4591 (2015). doi:10.1039/c4sm02718c

Z. Hong, K. J. Reeves, Z. Sun, Z. Li, N. J. Brown, and G. A. Meininger, "Vascular Smooth Muscle Cell Stiffness and Adhesion to Collagen I Modified by Vasoactive Agonists," PLOS ONE 10, e0119533 (2015). doi:10.1371/journal.pone.0119533

Z. Hong, K. J. Reeves, Z. Sun, Z. Li, N. J. Brown, and G. A. Meininger, "Vascular Smooth Muscle Cell Stiffness and Adhesion to Collagen I Modified by Vasoactive Agonists," PLOS ONE 10, e0119533 (2015). doi:10.1371/journal.pone.0119533

I. L. Ivanovska, J.-W. Shin, J. Swift, and D. E. Discher, "Stem cell mechanobiology: diverse lessons from bone marrow," Trends Cell Biol. 25, 523-532 (2015). doi:10.1016/j.tcb.2015.04.003

J. Jaczewska, M. H. Abdulreda, C. Y. Yau, M. M. Schmitt, I. Schubert, P.-O. Berggren, C. Weber, R. R. Koenen, V. T. Moy, and E. P. Wojcikiewicz, "TNF-α and IFN-γ promote lymphocyte adhesion to endothelial junctional regions facilitating transendothelial migration," J. Leukocyte Biol. 95, 265-274 (2013). doi:10.1189/jlb.0412205

H. Jin, D. A. Heller, M. Kalbacova, J.-H. Kim, J. Zhang, A. A. Boghossian, N. Maheshri, and M. S. Strano, "Detection of single-molecule H2O2 signalling from epidermal growth factor receptor using fluorescent single-walled carbon nanotubes," Nat. Nanotechnol. 5, 302-309 (2010). doi:10.1038/nnano.2010.24

G. Kaushik, A. C. Zambon, A. Fuhrmann, S. I. Bernstein, R. Bodmer, A. J. Engler, and A. Cammarato, "Measuring passive myocardial stiffness in Drosophila melanogaster to investigate diastolic dysfunction," J. Cell. Mol. Med. 16, 1656-1662 (2012). doi:10.1111/j.1582-4934.2011.01517.x

M. S. Kellermayer, Á. Karsai, A. Kengyel, A. Nagy, P. Bianco, T. Huber, Á. Kulcsár, C. Niedetzky, R. Proksch, and L. Grama, "Spatially and Temporally Synchronized Atomic Force and Total Internal Reflection Fluorescence Microscopy for Imaging and Manipulating Cells and Biomolecules," Biophys. J.91, 2665-2677 (2006). doi:10.1529/biophysj.106.085456

J. Liu, N. Sun, M. A. Bruce, J. C. Wu, and M. J. Butte, "Atomic Force Mechanobiology of Pluripotent Stem Cell-Derived Cardiomyocytes," PLoS ONE 7, e37559 (2012). doi:10.1371/journal.pone.0037559

J. I. Lopez, I. Kang, W.-K. You, D. M. McDonald, and V. M. Weaver, "In situ force mapping of mammary gland transformation," Integr. Biol. 3, 910 (2011). doi:10.1039/c1ib00043h

J. L. Maciaszek, and G. Lykotrafitis, "Sickle cell trait human erythrocytes are significantly stiffer than normal," J. Biomech. 44, 657-661 (2011). doi:10.1016/j.jbiomech.2010.11.008

J. M. Maloney, D. Nikova, F. Lautenschläger, E. Clarke, R. Langer, J. Guck, and K. J. V. Vliet, "Mesenchymal Stem Cell Mechanics from the Attached to the Suspended State," Biophys. J. 99, 2479-2487 (2010). doi:10.1016/j.bpj.2010.08.052

J. K. Mouw, Y. Yui, L. Damiano, R. O. Bainer, J. N. Lakins, I. Acerbi, G. Ou, A. C. Wijekoon, K. R. Levental, P. M. Gilbert, E. S. Hwang, Y.-Y. Chen, and V. M. Weaver, "Tissue mechanics modulate microRNA-dependent PTEN expression to regulate malignant progression," Nat. Med. 20, 360-367 (2014). doi:10.1038/nm.3497

M. F. Murphy, M. J. Lalor, F. C. Manning, F. Lilley, S. R. Crosby, C. Randall, and D. R. Burton, "Comparative study of the conditions required to image live human epithelial and fibroblast cells using atomic force microscopy," Microsc. Res. Tech. 69, 757-765 (2006). doi:10.1002/jemt.20339

M. Prabhune, G. Belge, A. Dotzauer, J. Bullerdiek, and M. Radmacher, "Comparison of mechanical properties of normal and malignant thyroid cells," Micron 43, 1267-1272 (2012). doi:10.1016/j.micron.2012.03.023

M. Prass, "Direct measurement of the lamellipodial protrusive force in a migrating cell," J. Cell Biol. 174, 767-772 (2006). doi:10.1083/jcb.200601159

A. Raman, S. Trigueros, A. Cartagena, A. P. Z. Stevenson, M. Susilo, E. Nauman, and S. A. Contera, "Mapping nanomechanical properties of live cells using multi-harmonic atomic force microscopy," Nat. Nanotechnol. 6, 809-814 (2011). doi:10.1038/nnano.2011.186

F. Rehfeldt, A. E. X. Brown, M. Raab, S. Cai, A. L. Zajac, A. Zemel, and D. E. Discher, "Hyaluronic acid matrices show matrix stiffness in 2D and 3D dictates cytoskeletal order and myosin-II phosphorylation within stem cells," Integr. Biol. 4, 422 (2012). doi:10.1039/c2ib00150k

J. Rother, H. Noding, I. Mey, and A. Janshoff, "Atomic force microscopy-based microrheology reveals significant differences in the viscoelastic response between malign and benign cell lines," Open Biology 4, 140046-140046 (2014). doi:10.1098/rsob.140046

S. Sen, S. Subramanian, and D. E. Discher, "Indentation and Adhesive Probing of a Cell Membrane with AFM: Theoretical Model and Experiments," Biophys. J. 89, 3203-3213 (2005). doi:10.1529/biophysj.105.063826

E. Spedden, and C. Staii, "Neuron Biomechanics Probed by Atomic Force Microscopy," Int. J. Mol. Sci.14, 16124-16140 (2013). doi:10.3390/ijms140816124

E. Spedden, J. D. White, E. N. Naumova, D. L. Kaplan, and C. Staii, "Elasticity Maps of Living Neurons Measured by Combined Fluorescence and Atomic Force Microscopy," Biophys. J. 103, 868-877 (2012). doi:10.1016/j.bpj.2012.08.005

J. R. Tse, and A. J. Engler, "Stiffness Gradients Mimicking In Vivo Tissue Variation Regulate Mesenchymal Stem Cell Fate," PLoS ONE 6, e15978 (2011). doi:10.1371/journal.pone.0015978

K. R. Wilhelm, E. Roan, M. C. Ghosh, K. Parthasarathi, and C. M. Waters, "Hyperoxia increases the elastic modulus of alveolar epithelial cells through Rho kinase," FEBS Journal 281, 957-969 (2013). doi:10.1111/febs.12661

Y. Xiong, A. C. Lee, D. M. Suter, and G. U. Lee, "Topography and Nanomechanics of Live Neuronal Growth Cones Analyzed by Atomic Force Microscopy," Biophys. J. 96, 5060-5072 (2009). doi:10.1016/j.bpj.2009.03.032

W. Xu, R. Mezencev, B. Kim, L. Wang, J. McDonald, and T. Sulchek, "Cell Stiffness Is a Biomarker of the Metastatic Potential of Ovarian Cancer Cells," PLoS ONE 7, e46609 (2012). doi:10.1371/journal.pone.0046609

E. K. Yim, E. M. Darling, K. Kulangara, F. Guilak, and K. W. Leong, "Nanotopography-induced changes in focal adhesions, cytoskeletal organization, and mechanical properties of human mesenchymal stem cells," Biomaterials 31, 1299-1306 (2010). doi:10.1016/j.biomaterials.2009.10.037

【セミナー開催】 EBSDセミナーと講習会追加日程のお知らせ https://t.co/HsrIcsNEVV
1:50 午後 - 6 7 17
さらにTweetを見る