Skip to main content

Imaging Living Chondrocyte Surface Structures With AFM Contact Mode

  • Protocol
Book cover Atomic Force Microscopy

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 242))

  • 1634 Accesses

Abstract

In its most established mode of operation, named constant force contact mode, atomic force microscopy (AFM) has been applied to image the 2D and 3D architecture of surfaces. Any deflection of the tip as a result of surface topography is recorded. The microscope reconstructs an image of the surface from the x, y, and z scan data to develop a 3D illustration of any surface at the micro- and nanometer level. The production of high-resolution images of a wide variety of biological samples at near-native conditions and the possibility to measure very low local forces is proving to be a powerful tool for cell analysis (1,2). In contrast with electron microscopy observations in particular, AFM improves biological studies involving imaging by also monitoring dynamic processes. However, the investigation of soft biomaterials with this special method is still challenging. This chapter reviews practical details of imaging two cell lines: human chondrocytes and human osteosarcoma. However, characteristics described are not unique to this type of cell. Principally, all types of adherently growing cells can be investigated with the techniques described here. Force curve analysis, as a backdrop for the understanding of the received images (1), will be introduced in detail in Subheading 3.4. Further sections explore how AFM can be used as a helpful tool in observations of the cell surface and the physical interactions that occur there, like adhesion or friction, and their influence on the active cell. In Subheading 7. common artifacts and troubles are described, along with the practical instructions.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Ricci, D. and Grattarola, M. (1994) Scanning force microscopy on live cultured cells: imaging and force-versus-distance investigations. J. Microscopy 176, 254–261.

    CAS  Google Scholar 

  2. Oberleithner, H., Brinckmann, E., Giebisch, G., and Geibel, J. (1995) Visualizing life on biomembranes by atomic force microscopy. Kidney Int. 48, 923–929.

    Article  PubMed  CAS  Google Scholar 

  3. Wohlrab, D., Wohlrab, J., Reichel, H., and Hein, W. (2001) Is the proliferation of human chondrocytes regulated by ionic channels? J. Orthop. Sci. 6, 155–159.

    Article  PubMed  CAS  Google Scholar 

  4. Grodzinsky, A. J., Levenston, M. E., Jin, M., and Frank, E. H. (2000) Cartilage tissue remodeling in response to mechanical forces. Annu. Rev. Biomed. Eng. 2, 691–713.

    Article  PubMed  CAS  Google Scholar 

  5. Hein, H.-J., Brandt, J., Bernstein, A., Engler, T., and Weisser, L. (1997) Zur Darstellung der Mikrostruktur des Knochens mit dem Raster-Sondenmikroskop. Z. Med. Phys. 7, 21–26.

    Google Scholar 

  6. Henning, S., Adhikari, R., Michler, G. H., Seidel, P., Sandner, B., Bernstein, A., and Hein, H.-J. (2001) Analysis of the bone-implant interface of a partially resorbable bone cement by scanning electron and scanning force microscopy, in Micro-and Nanostructures of Biological Systems, (Bischoff, G. and Hein, H.-J., eds.), Shaker-Publ., Aachen, pp. 109–120.

    Google Scholar 

  7. Oda, Y., Matsumoto, Y., Harimaya, K., Iwamoto, Y., and Tsuneyoshi, M. (2000) Establishment of new multidrug-resistant human osteosarcoma cell lines. Oncol-Rep. 7, 859–866.

    PubMed  CAS  Google Scholar 

  8. Rodan, S. B., Imai, Y., Thiede, M. A., Wesolowski, G., Thompson, D., Bar-Shavit, Z., et al. (1987) Characterization of a human osteosarcoma cell line (Saos-2) with osteoblastic properties. Cancer Res. 47, 4961–4966.

    PubMed  CAS  Google Scholar 

  9. Rhim, J. S. (1993) Neoplastic transformation of human cells in vitro. Crit. Rev. Oncogene 4, 313–335.

    CAS  Google Scholar 

  10. Sakai, K., Mohtai, M., and Iwamoto, Y. (1998) Fluid shear stress increases transforming growth factor beta 1 expression in human osteoblast-like cells: modulation by cation channel blockades. Calcif. Tissue Int. 63, 515–520.

    Article  PubMed  CAS  Google Scholar 

  11. Grimsehl, E., (1989) Lehrbuch der Physik Vol. 1: Mechanik, Akustik, Wärmelehre, 25th ed. Teubners-Verlagsgesellschaft, Leipzig. pp. 43–56.

    Google Scholar 

  12. Takano, H., Kenseth, J. R., Wong, S.-S., O’Brien, J. C., and Porter, M. D. (1999) Chemical and biochemical analysis using scanning force microscopy. Chem. Rev. 99, 2845–2890.

    Article  PubMed  CAS  Google Scholar 

  13. Hansma, H. G. (2001) Surface biology of DNA by atomic force microscopy. Annu. Rev. Phys. Chem. 52, 71–92.

    Article  PubMed  CAS  Google Scholar 

  14. Linder, A., Weiland, U., and Apell, H. J. (1999) Novel polymer substrates for SFM investigations of living cells, biological membranes, and proteins. J. Struct. Biol. 126, 16–26.

    Article  PubMed  CAS  Google Scholar 

  15. Bischoff, R., Berghaus, A., and Hein, H.-J. (1997) Inspection of silicone-biomaterials using SPM. Biomed. Techn. 42(Suppl. 2), 482–483.

    Article  Google Scholar 

  16. Bustamante, C. and Keller D (1995) Scanning force microscopy in biology. Physics Today 32–38.

    Google Scholar 

  17. Benoit, M., Holstein, T., and Gaub, H. E. (1997) Lateral forces in AFM imaging and immobilization of cells and organelles. Eur. Biophys. J. 26, 283–290.

    Article  Google Scholar 

  18. Bischoff, G. and Langner, J. (2001) SFM of living cells—a study of the method, in Micro-and Nanostructures of Biological Systems, (Bischoff, G. and Hein, H.-J., eds), Shaker Publ., Aachen, pp. 135–152.

    Google Scholar 

  19. Bischoff, R., Bischoff, G., and Hein, H.-J. (2002) Scanning force microscopy (SFM) visualization of adherently growing cells. Am. Biotech. Lab. 3, 20–22.

    Google Scholar 

  20. Bischoff, R., Bischoff, G., and Hoffmann, S. (2001) Scanning force microscopy observation of tumor cells treated with hematoporphyrin IX derivatives. Ann. Biomed. Eng. 29, 1092–1099.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2004 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Bischoff, G., Bernstein, A., Wohlrab, D., Hein, HJ. (2004). Imaging Living Chondrocyte Surface Structures With AFM Contact Mode. In: Braga, P.C., Ricci, D. (eds) Atomic Force Microscopy. Methods in Molecular Biology™, vol 242. Humana Press. https://doi.org/10.1385/1-59259-647-9:105

Download citation

  • DOI: https://doi.org/10.1385/1-59259-647-9:105

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-094-6

  • Online ISBN: 978-1-59259-647-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics