Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 1998 Mar;74(3):1564–1578. doi: 10.1016/S0006-3495(98)77868-3

Relative microelastic mapping of living cells by atomic force microscopy.

E A-Hassan 1, W F Heinz 1, M D Antonik 1, N P D'Costa 1, S Nageswaran 1, C A Schoenenberger 1, J H Hoh 1
PMCID: PMC1299502  PMID: 9512052

Abstract

The spatial and temporal changes of the mechanical properties of living cells reflect complex underlying physiological processes. Following these changes should provide valuable insight into the biological importance of cellular mechanics and their regulation. The tip of an atomic force microscope (AFM) can be used to indent soft samples, and the force versus indentation measurement provides information about the local viscoelasticity. By collecting force-distance curves on a time scale where viscous contributions are small, the forces measured are dominated by the elastic properties of the sample. We have developed an experimental approach, using atomic force microscopy, called force integration to equal limits (FIEL) mapping, to produce robust, internally quantitative maps of relative elasticity. FIEL mapping has the advantage of essentially being independent of the tip-sample contact point and the cantilever spring constant. FIEL maps of living Madine-Darby canine kidney (MDCK) cells show that elasticity is uncoupled from topography and reveal a number of unexpected features. These results present a mode of high-resolution visualization in which the contrast is based on the mechanical properties of the sample.

Full Text

The Full Text of this article is available as a PDF (727.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Armitage W. J., Juss B. K., Easty D. L. Response of epithelial (MDCK) cell junctions to calcium removal and osmotic stress is influenced by temperature. Cryobiology. 1994 Oct;31(5):453–460. doi: 10.1006/cryo.1994.1055. [DOI] [PubMed] [Google Scholar]
  2. Ashkin A., Dziedzic J. M. Internal cell manipulation using infrared laser traps. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7914–7918. doi: 10.1073/pnas.86.20.7914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Balkovetz D. F., Pollack A. L., Mostov K. E. Hepatocyte growth factor alters the polarity of Madin-Darby canine kidney cell monolayers. J Biol Chem. 1997 Feb 7;272(6):3471–3477. doi: 10.1074/jbc.272.6.3471. [DOI] [PubMed] [Google Scholar]
  4. Bereiter-Hahn J., Karl I., Lüers H., Vöth M. Mechanical basis of cell shape: investigations with the scanning acoustic microscope. Biochem Cell Biol. 1995 Jul-Aug;73(7-8):337–348. doi: 10.1139/o95-042. [DOI] [PubMed] [Google Scholar]
  5. Binnig G, Quate CF, Gerber C. Atomic force microscope. Phys Rev Lett. 1986 Mar 3;56(9):930–933. doi: 10.1103/PhysRevLett.56.930. [DOI] [PubMed] [Google Scholar]
  6. Butt H. J., Wolff E. K., Gould S. A., Dixon Northern B., Peterson C. M., Hansma P. K. Imaging cells with the atomic force microscope. J Struct Biol. 1990 Oct-Dec;105(1-3):54–61. doi: 10.1016/1047-8477(90)90098-w. [DOI] [PubMed] [Google Scholar]
  7. Dugina V. B., Alexandrova A. Y., Lane K., Bulanova E., Vasiliev J. M. The role of the microtubular system in the cell response to HGF/SF. J Cell Sci. 1995 Apr;108(Pt 4):1659–1667. doi: 10.1242/jcs.108.4.1659. [DOI] [PubMed] [Google Scholar]
  8. Goldmann W. H., Ezzell R. M. Viscoelasticity in wild-type and vinculin-deficient (5.51) mouse F9 embryonic carcinoma cells examined by atomic force microscopy and rheology. Exp Cell Res. 1996 Jul 10;226(1):234–237. doi: 10.1006/excr.1996.0223. [DOI] [PubMed] [Google Scholar]
  9. Hansma H. G., Hoh J. H. Biomolecular imaging with the atomic force microscope. Annu Rev Biophys Biomol Struct. 1994;23:115–139. doi: 10.1146/annurev.bb.23.060194.000555. [DOI] [PubMed] [Google Scholar]
  10. Haydon P. G., Lartius R., Parpura V., Marchese-Ragona S. P. Membrane deformation of living glial cells using atomic force microscopy. J Microsc. 1996 May;182(Pt 2):114–120. doi: 10.1046/j.1365-2818.1996.141423.x. [DOI] [PubMed] [Google Scholar]
  11. Henderson E., Haydon P. G., Sakaguchi D. S. Actin filament dynamics in living glial cells imaged by atomic force microscopy. Science. 1992 Sep 25;257(5078):1944–1946. doi: 10.1126/science.1411511. [DOI] [PubMed] [Google Scholar]
  12. Hoh J. H., Schoenenberger C. A. Surface morphology and mechanical properties of MDCK monolayers by atomic force microscopy. J Cell Sci. 1994 May;107(Pt 5):1105–1114. doi: 10.1242/jcs.107.5.1105. [DOI] [PubMed] [Google Scholar]
  13. Huotari V., Vaaraniemi J., Lehto V. P., Eskelinen S. Regulation of the disassembly/assembly of the membrane skeleton in Madin-Darby canine kidney cells. J Cell Physiol. 1996 Apr;167(1):121–130. doi: 10.1002/(SICI)1097-4652(199604)167:1<121::AID-JCP14>3.0.CO;2-E. [DOI] [PubMed] [Google Scholar]
  14. Ingber D. E. Cellular tensegrity: defining new rules of biological design that govern the cytoskeleton. J Cell Sci. 1993 Mar;104(Pt 3):613–627. doi: 10.1242/jcs.104.3.613. [DOI] [PubMed] [Google Scholar]
  15. Ingber D. E., Dike L., Hansen L., Karp S., Liley H., Maniotis A., McNamee H., Mooney D., Plopper G., Sims J. Cellular tensegrity: exploring how mechanical changes in the cytoskeleton regulate cell growth, migration, and tissue pattern during morphogenesis. Int Rev Cytol. 1994;150:173–224. doi: 10.1016/s0074-7696(08)61542-9. [DOI] [PubMed] [Google Scholar]
  16. Ingber D. E., Prusty D., Sun Z., Betensky H., Wang N. Cell shape, cytoskeletal mechanics, and cell cycle control in angiogenesis. J Biomech. 1995 Dec;28(12):1471–1484. doi: 10.1016/0021-9290(95)00095-x. [DOI] [PubMed] [Google Scholar]
  17. Laney D. E., Garcia R. A., Parsons S. M., Hansma H. G. Changes in the elastic properties of cholinergic synaptic vesicles as measured by atomic force microscopy. Biophys J. 1997 Feb;72(2 Pt 1):806–813. doi: 10.1016/s0006-3495(97)78714-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Li M. L., Aggeler J., Farson D. A., Hatier C., Hassell J., Bissell M. J. Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mouse mammary epithelial cells. Proc Natl Acad Sci U S A. 1987 Jan;84(1):136–140. doi: 10.1073/pnas.84.1.136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lüers H., Hillmann K., Litniewski J., Bereiter-Hahn J. Acoustic microscopy of cultured cells. Distribution of forces and cytoskeletal elements. Cell Biophys. 1991 Jun;18(3):279–293. doi: 10.1007/BF02989819. [DOI] [PubMed] [Google Scholar]
  20. Maniotis A. J., Chen C. S., Ingber D. E. Demonstration of mechanical connections between integrins, cytoskeletal filaments, and nucleoplasm that stabilize nuclear structure. Proc Natl Acad Sci U S A. 1997 Feb 4;94(3):849–854. doi: 10.1073/pnas.94.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mitchison T. J. Evolution of a dynamic cytoskeleton. Philos Trans R Soc Lond B Biol Sci. 1995 Sep 29;349(1329):299–304. doi: 10.1098/rstb.1995.0117. [DOI] [PubMed] [Google Scholar]
  22. Mooney D., Hansen L., Vacanti J., Langer R., Farmer S., Ingber D. Switching from differentiation to growth in hepatocytes: control by extracellular matrix. J Cell Physiol. 1992 Jun;151(3):497–505. doi: 10.1002/jcp.1041510308. [DOI] [PubMed] [Google Scholar]
  23. Pasdar M., Li Z. Disorganization of microfilaments and intermediate filaments interferes with the assembly and stability of desmosomes in MDCK epithelial cells. Cell Motil Cytoskeleton. 1993;26(2):163–180. doi: 10.1002/cm.970260207. [DOI] [PubMed] [Google Scholar]
  24. Petersen N. O., McConnaughey W. B., Elson E. L. Dependence of locally measured cellular deformability on position on the cell, temperature, and cytochalasin B. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5327–5331. doi: 10.1073/pnas.79.17.5327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Putman C. A., van der Werf K. O., de Grooth B. G., van Hulst N. F., Greve J. Viscoelasticity of living cells allows high resolution imaging by tapping mode atomic force microscopy. Biophys J. 1994 Oct;67(4):1749–1753. doi: 10.1016/S0006-3495(94)80649-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Radmacher M., Cleveland J. P., Fritz M., Hansma H. G., Hansma P. K. Mapping interaction forces with the atomic force microscope. Biophys J. 1994 Jun;66(6):2159–2165. doi: 10.1016/S0006-3495(94)81011-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Radmacher M., Fritz M., Hansma P. K. Imaging soft samples with the atomic force microscope: gelatin in water and propanol. Biophys J. 1995 Jul;69(1):264–270. doi: 10.1016/S0006-3495(95)79897-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Radmacher M., Fritz M., Kacher C. M., Cleveland J. P., Hansma P. K. Measuring the viscoelastic properties of human platelets with the atomic force microscope. Biophys J. 1996 Jan;70(1):556–567. doi: 10.1016/S0006-3495(96)79602-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schaus S. S., Henderson E. R. Cell viability and probe-cell membrane interactions of XR1 glial cells imaged by atomic force microscopy. Biophys J. 1997 Sep;73(3):1205–1214. doi: 10.1016/S0006-3495(97)78153-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schoenenberger C. A., Hoh J. H. Slow cellular dynamics in MDCK and R5 cells monitored by time-lapse atomic force microscopy. Biophys J. 1994 Aug;67(2):929–936. doi: 10.1016/S0006-3495(94)80556-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shao J. Y., Hochmuth R. M. Micropipette suction for measuring piconewton forces of adhesion and tether formation from neutrophil membranes. Biophys J. 1996 Nov;71(5):2892–2901. doi: 10.1016/S0006-3495(96)79486-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shroff S. G., Saner D. R., Lal R. Dynamic micromechanical properties of cultured rat atrial myocytes measured by atomic force microscopy. Am J Physiol. 1995 Jul;269(1 Pt 1):C286–C292. doi: 10.1152/ajpcell.1995.269.1.C286. [DOI] [PubMed] [Google Scholar]
  33. Singhvi R., Kumar A., Lopez G. P., Stephanopoulos G. N., Wang D. I., Whitesides G. M., Ingber D. E. Engineering cell shape and function. Science. 1994 Apr 29;264(5159):696–698. doi: 10.1126/science.8171320. [DOI] [PubMed] [Google Scholar]
  34. Svoboda K., Schmidt C. F., Branton D., Block S. M. Conformation and elasticity of the isolated red blood cell membrane skeleton. Biophys J. 1992 Sep;63(3):784–793. doi: 10.1016/S0006-3495(92)81644-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tao N. J., Lindsay S. M., Lees S. Measuring the microelastic properties of biological material. Biophys J. 1992 Oct;63(4):1165–1169. doi: 10.1016/S0006-3495(92)81692-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Valberg P. A., Feldman H. A. Magnetic particle motions within living cells. Measurement of cytoplasmic viscosity and motile activity. Biophys J. 1987 Oct;52(4):551–561. doi: 10.1016/S0006-3495(87)83244-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wacker I. U., Rickard J. E., De Mey J. R., Kreis T. E. Accumulation of a microtubule-binding protein, pp170, at desmosomal plaques. J Cell Biol. 1992 May;117(4):813–824. doi: 10.1083/jcb.117.4.813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wang N., Ingber D. E. Control of cytoskeletal mechanics by extracellular matrix, cell shape, and mechanical tension. Biophys J. 1994 Jun;66(6):2181–2189. doi: 10.1016/S0006-3495(94)81014-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Weisenhorn AL, Maivald P, Butt H, Hansma PK. Measuring adhesion, attraction, and repulsion between surfaces in liquids with an atomic-force microscope. Phys Rev B Condens Matter. 1992 May 15;45(19):11226–11232. doi: 10.1103/physrevb.45.11226. [DOI] [PubMed] [Google Scholar]
  40. Yeung A., Evans E. Cortical shell-liquid core model for passive flow of liquid-like spherical cells into micropipets. Biophys J. 1989 Jul;56(1):139–149. doi: 10.1016/S0006-3495(89)82659-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zahalak G. I., McConnaughey W. B., Elson E. L. Determination of cellular mechanical properties by cell poking, with an application to leukocytes. J Biomech Eng. 1990 Aug;112(3):283–294. doi: 10.1115/1.2891186. [DOI] [PubMed] [Google Scholar]
  42. Zeman K., Engelhard H., Sackmann E. Bending undulations and elasticity of the erythrocyte membrane: effects of cell shape and membrane organization. Eur Biophys J. 1990;18(4):203–219. doi: 10.1007/BF00183373. [DOI] [PubMed] [Google Scholar]
  43. Zheng J., Lamoureux P., Santiago V., Dennerll T., Buxbaum R. E., Heidemann S. R. Tensile regulation of axonal elongation and initiation. J Neurosci. 1991 Apr;11(4):1117–1125. doi: 10.1523/JNEUROSCI.11-04-01117.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES