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Influence of polylysine on adhesion of fibroblasts to glass substrates visualized by total internal reflection microscopy

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EBO — Experimental Biology Online Annual 1996/97

Part of the book series: EBO — Experimental Biology Online Annual ((EBOEXP,volume 1996/1997))

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Abstract

The cell adhesion topography of mouse fibroblasts growing on glass substrates has been investigated. In order to compare cell adhesion on covered and uncovered glass, substrates were partly exposed to a solution with 0.1 mg/ml polylysine (300 kDa) for 15 min before incubation with cell suspension. After cultivation for 1, 3, 6, and 24 h their adhesion was visualised by total internal reflection microscopy. In the presence of polylysine, cells incubated for 1 h were strongly attracted to the substrate, leading to a typical cell adhesion topography characterised by round concavities under the ventral cell membrane with an approximate diameter of 1 μm. The cavity-surrounding rims were tightly bound to the glass surface. During further cell cultivation, the topography changed into a well-organised adhesion pattern with focal contact areas on the periphery of the cells. In contrast to the polylysine-mediated adhesion, cells growing on untreated surfaces did not exhibit the cavity-like topography at any stage of cultivation, but a more point spread adhesion with a dense clustering of contact-forming areas.

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References

  • Axelrod, D. (1981) Cell-substrate contacts illuminated by Total Internal Reflection Fluorescence. J. Cell Biol. 89: 141–145

    Article  PubMed  CAS  Google Scholar 

  • Bereiter-Hahn, J., Fox, C.H., Thorell, B. (1979) Quantitative reflection contrast microscopy of living cells. J. Cell Biol. 82: 767–779

    Article  PubMed  CAS  Google Scholar 

  • Bereiter-Hahn, J., Lück, M., Miebach, T., Stelzer, H.K., Vöth, M. (199o) Spreading of trypsinized cells: cytoskeletal dynamics and energy requirements. J. Cell Sci. 96: 171–188

    Google Scholar 

  • Bray, D., Heath, J., Moss, D. (1986) The membrane-associated “cortex” of animal cells: Its structure and mechanical properties J. Cell Sci. (Suppl.) 4: 71–88

    CAS  Google Scholar 

  • Burmeister, J.S., Truskey, G.A., Reichert, W.M. (1994) Quantitative analysis of variable-angle total internal reflection fluorescence microscopy (VA-TIRFM) of cell/substrate contacts. J. Microsc. 173: 39–51

    Google Scholar 

  • Curtis, A.S.G. (1964) The mechanism of adhesion of cells to glass. A study by interference reflection microscopy. J. Cell Biol. 20: 199–215

    Article  PubMed  CAS  Google Scholar 

  • Gingell, D., Heavens, O.S. (1996) Elimination of the effects of stray light in measurements by total internal reflection aqueous fluorescence (TIRAF). J. Microsc. 182: 141–148

    Article  PubMed  CAS  Google Scholar 

  • Gingell, D., Todd, I. (1979) Interference reflection microscopy. A quantitative theory for image interpretation and its application to cell-substratum separation measurement. Biophys. J. 26: 507–526

    Article  PubMed  CAS  Google Scholar 

  • Gingell, D., Todd, I., Bailey, J. (1985) Topography of cell-glass apposition revealed by Total Internal Reflection Fluorescence of Volume Markers. J. Cell

    Google Scholar 

  • Biol. 100: 1334–1338

    Google Scholar 

  • Gingell, D., Heavens, O.S., Mellor, J.S. (1987) General electromagnetic theory of total internal reflection fluorescence: the quantitative basis for mapping cell-substratum topography. J. Cell Sci. 87: 677–693

    PubMed  Google Scholar 

  • Heavens, O.S., Gingell, D. (1991) Film thickness measurement by frustrated total reflection fluorescence. Opt. Laser Technol. 23: 175–179

    Article  CAS  Google Scholar 

  • Hornung, J., Müller, T., Fuhr, G. (1996) Cryopreservation of anchorage-dependent mammalian cells fixed to structured glass and silicon substrates. Cryobiology 33: 260–270

    Article  PubMed  CAS  Google Scholar 

  • Ireland, G.W., Dopping-Hepenstal, P.J.C., Jordan, P.W., O’Neill, C.H. (1989) Limitation of substratum size alters cytoskeletal organization and behaviour of swiss 3T3 fibroblast. Cell Biol. Int. 13: 781–790

    Article  CAS  Google Scholar 

  • Izzard, C.S., Lochner, L.R. (1976) Cell-to-substrate contacts in living fibroblasts: an interference reflection study with an evaluation of the technique. J. Cell Sci. 21: 129–159

    PubMed  CAS  Google Scholar 

  • Lanni, F., Waggoner, A.S., Taylor, D.L. (1985) Structural organization of interphase 3T3 fibroblasts studied by total internal reflection fluorescence microscopy. J. Cell Biol. 100: 1091–1102

    Article  PubMed  CAS  Google Scholar 

  • Maurer, H.R. (1992) Towards serum-free, chemically defined media for mammalian cell culture. In: Freshney, R.I. (ed.) Animal cell culture: a practical approach., Oxford University Press, Oxford, pp. 15–46

    Google Scholar 

  • Meisingset, K.K., Steen, H.B (1981) Intracellular bindings of fluorescein in lymphocytes. Cytometry 1: 272–278

    Article  PubMed  CAS  Google Scholar 

  • Thomas, N.E., Coakley, W.T., Galip, A. (1992) The lateral separation of contacts on erythrocytes agglutinated by polylysine. Cell Biophys. 20: 125–147

    PubMed  CAS  Google Scholar 

  • Thomas, N.E., Coakley, W.T., Winters, C. (1996) Contact formation in polylysine-mediated membrane-glass interaction. Colloids Surfaces B. 6: 139–147

    Article  CAS  Google Scholar 

  • Truskey, G.A., Burmeister, J.S., Grapa, E., Reichert, W.M. (1992) Total internal reflection fluorescence microscopy (TIRFM). II. Topographical mapping of relative cell/substratum separation distances. J. Cell Sci. 103: 491–499’

    PubMed  Google Scholar 

  • Tsien, R.Y. (1989) Fluorescent indicators of ion concentrations. In: Taylor, D.L., Wang, Y. (eds.) Fluorescence microscopy of living cells in culture: part B., Academic Press, San Diego, pp. 127–156

    Chapter  Google Scholar 

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© 1998 Springer-Verlag Berlin Heidelberg

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Hornung, J., Fuhr, G. (1998). Influence of polylysine on adhesion of fibroblasts to glass substrates visualized by total internal reflection microscopy. In: EBO — Experimental Biology Online Annual 1996/97. EBO — Experimental Biology Online Annual, vol 1996/1997. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-00932-1_10

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  • DOI: https://doi.org/10.1007/978-3-662-00932-1_10

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-662-00934-5

  • Online ISBN: 978-3-662-00932-1

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