Histochemistry and Cell Biology

, Volume 122, Issue 3, pp 183–190

Dynamic imaging of cellular interactions with extracellular matrix

Histochemistry and Cell Biology Lecture

Abstract

Adhesive and proteolytic interactions of cells with components of the extracellular matrix (ECM) are fundamental to morphogenesis, tissue assembly and remodeling, and cell migration as well as signal acquisition from tissue-bound factors. The visualization from fixed samples provides snapshot-like, static information on the cellular and molecular dynamics of adhesion receptor and protease functions toward ECM, such as interstitial fibrillar tissues and basement membranes. Recent technological developments additionally support the dynamic imaging of ECM scaffolds and the interaction behavior of cells contained therein. These include differential interference contrast, confocal reflection microscopy, optical coherence tomography, and multiphoton microscopy and second-harmonic generation imaging. Most of these approaches are combined with fluorescence imaging using derivates of GFP and/or other fluorescent dyes. Dynamic 3D imaging has revealed an unexpected degree of dynamics and turnover of cell adhesion and migration as well as basic mechanisms that lead to proteolytic remodeling of connective tissue by stromal cells and invading tumor cells.

Keywords

Collagen matrix Intravital microscopy Cell migration Integrins Attachment Detachment Tissue remodeling Tumor invasion 

Supplementary material

Movie 1 Time-resolved confocal reflection and fluorescence imaging of MV3 melanoma cell migrating within 3D collagen lattice. Traction of collagen fibers by the cell and forward movement are accompanied by the release of CD44 from the trailing edge upon cell detachment. Cells were labeled with anti-CD44 mAb Hermes-3 and secondary non-cross-linking LRSC-conjugated goat-anti mouse F(ab)’- fragments, thus avoiding receptor hypercrosslinking. Images of the fluorescent (red) and reflection channel (grayscale) were obtained every 5 min and displayed in false-color. Time is indicated in the lower right corner.

MV3CD44c3.avi (1.8 mb)
MV3CD44c3.avi (1.8 MB)

References

  1. Aletta JM, Greene LA (1988) Growth cone configuration and advance: a time-lapse study using video-enhanced differential interference contrast microscopy. J Neurosci 8:1425–1435PubMedGoogle Scholar
  2. Barton JK, Gossage KW, Xu W, Ranger-Moore JR, Saboda K, Brooks CA, Duckett LD, Salasche SJ, Warneke JA, Alberts DS (2003) Investigating sun-damaged skin and actinic keratosis with optical coherence tomography: a pilot study. Technol Cancer Res Treat 2:525–535PubMedGoogle Scholar
  3. Boyde A, Jones SJ (1995) Mapping and measuring surfaces using reflection confocal microscopy. In: Pawley JB (ed) Handbook of biological confocal microscopy. Plenum, New York, pp 255–265Google Scholar
  4. Brightman AO, Rajwa BP, Sturgis JE, McCallister ME, Robinson JP, Voytik-Harbin SL (2000) Time-lapse confocal reflection microscopy of collagen fibrillogenesis and extracellular matrix assembly in vitro. Biopolymers 54:222–234CrossRefPubMedGoogle Scholar
  5. Campagnola PJ, Loew LM (2003) Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms. Nat Biotechnol 21:1356–1360CrossRefPubMedGoogle Scholar
  6. Campagnola PJ, Millard AC, Terasaki M, Hoppe PE, Malone CJ, Mohler WA (2002) Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues. Biophys J 82:493–508PubMedGoogle Scholar
  7. Cheng PC, Kriete A (1995) Image contrast in confocal light microscopy. In: Pawley JB (ed) Handbook of biological confocal microscopy. Plenum, New York, pp 281–310Google Scholar
  8. Cukierman E, Pankov R, Stevens DR, Yamada KM (2001) Taking cell-matrix adhesions to the third dimension. Science 294:1708–1712CrossRefPubMedGoogle Scholar
  9. Davies PF, Robotewskyj A, Griem ML (1993) Endothelial cell adhesion in real time. Measurements in vitro by tandem scanning confocal image analysis. J Clin Invest 91:2640–2652PubMedGoogle Scholar
  10. Denk W, Strickler JH, Webb WW (1990) Two-photon laser scanning fluorescence microscopy. Science 248:73–76PubMedGoogle Scholar
  11. Dickinson ME, Simbuerger E, Zimmermann B, Waters CW, Fraser SE (2003) Multiphoton excitation spectra in biological samples. J Biomed Opt 8:329–338CrossRefPubMedGoogle Scholar
  12. Dombeck DA, Kasischke KA, Vishwasrao HD, Ingelsson M, Hyman BT, Webb WW (2003) Uniform polarity microtubule assemblies imaged in native brain tissue by second-harmonic generation microscopy. Proc Natl Acad Sci U S A 100:7081–7086CrossRefPubMedGoogle Scholar
  13. Friedl P (2004a) Dynamic imaging of the immune system. Curr Opin Immunol 16:389–393CrossRefGoogle Scholar
  14. Friedl P (2004b) Prespecification and plasticity: shifting mechanisms of cell migration. Curr Opin Cell Biol 16:14–23CrossRefPubMedGoogle Scholar
  15. Friedl P, Brocker EB (2004) Reconstructing leukocyte migration in 3D extracellular matrix by time-lapse videomicroscopy and computer-assisted tracking. Methods Mol Biol 239:77–90PubMedGoogle Scholar
  16. Friedl P, Wolf K (2003a) Proteolytic and non-proteolytic migration of tumour cells and leucocytes. Biochem Soc Symp 70:277–285PubMedGoogle Scholar
  17. Friedl P, Wolf K (2003b) Tumour-cell invasion and migration: diversity and escape mechanisms. Nat Rev Cancer 3:362–374CrossRefPubMedGoogle Scholar
  18. Friedl P, Maaser K, Klein CE, Niggemann B, Krohne G, Zanker KS (1997) Migration of highly aggressive MV3 melanoma cells in 3-dimensional collagen lattices results in local matrix reorganization and shedding of alpha2 and beta1 integrins and CD44. Cancer Res 57:2061–2070PubMedGoogle Scholar
  19. Friedl P, Entschladen F, Conrad C, Niggemann B, Zanker KS (1998a) CD4+ T lymphocytes migrating in three-dimensional collagen lattices lack focal adhesions and utilize beta1 integrin-independent strategies for polarization, interaction with collagen fibers and locomotion. Eur J Immunol 28:2331–2343CrossRefPubMedGoogle Scholar
  20. Friedl P, Zanker KS, Brocker EB (1998b) Cell migration strategies in 3-D extracellular matrix: differences in morphology, cell matrix interactions, and integrin function. Microsc Res Tech 43:369–378CrossRefPubMedGoogle Scholar
  21. Friedl P, Borgmann S, Brocker EB (2001) Amoeboid leukocyte crawling through extracellular matrix: lessons from the Dictyostelium paradigm of cell movement. J Leukoc Biol 70:491–509PubMedGoogle Scholar
  22. Friedl P, Hegerfeldt Y, Tusch M (2004) Collective cell migration in morphogenesis and cancer. Eur J Dev Biol (in press)Google Scholar
  23. Fukuchi T, Takahashi K, Uyama M, Matsumura M (2001) Comparative study of experimental choroidal neovascularization by optical coherence tomography and histopathology. Jpn J Ophthalmol 45:252–258CrossRefPubMedGoogle Scholar
  24. Gladkova ND, Petrova GA, Nikulin NK, Radenska-Lopovok SG, Snopova LB, Chumakov YP, Nasonova VA, Gelikonov VM, Gelikonov GV, Kuranov RV, Sergeev AM, Feldchtein FI (2000) In vivo optical coherence tomography imaging of human skin: norm and pathology. Skin Res Technol 6:6–16CrossRefPubMedGoogle Scholar
  25. Hegerfeldt Y, Tusch M, Brocker EB, Friedl P (2002) Collective cell movement in primary melanoma explants: plasticity of cell-cell interaction, beta1-integrin function, and migration strategies. Cancer Res 62:2125–2130PubMedGoogle Scholar
  26. Huang D, Swanson EA, Lin CP, Schuman JS, Stinson WG, Chang W, Hee MR, Flotte T, Gregory K, Puliafito CA, et al (1991) Optical coherence tomography. Science 254:1178–1181PubMedGoogle Scholar
  27. Keith CH, Bird GJ, Farmer MA (1998) Coherent backscatter enhances reflection confocal microscopy. Biotechniques 25:858–866PubMedGoogle Scholar
  28. Konig K (2000) Multiphoton microscopy in life sciences. J Microsc 200:83–104CrossRefPubMedGoogle Scholar
  29. Konig K, Riemann I (2003) High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution. J Biomed Opt 8:432–439CrossRefPubMedGoogle Scholar
  30. Korff T, Augustin HG (1999) Tensional forces in fibrillar extracellular matrices control directional capillary sprouting. J Cell Sci 112:3249–3258PubMedGoogle Scholar
  31. Lippincott-Schwartz J, Altan-Bonnet N, Patterson GH (2003) Photobleaching and photoactivation: following protein dynamics in living cells. Nat Cell Biol 5(suppl):S7–S14CrossRefGoogle Scholar
  32. Maaser K, Wolf K, Klein CE, Niggemann B, Zanker KS, Brocker EB, Friedl P (1999) Functional hierarchy of simultaneously expressed adhesion receptors: integrin alpha2beta1 but not CD44 mediates MV3 melanoma cell migration and matrix reorganization within three-dimensional hyaluronan-containing collagen matrices. Mol Biol Cell 10:3067–3079PubMedGoogle Scholar
  33. Mempel TR, Henrickson SE, Von Andrian UH (2004a) T-cell priming by dendritic cells in lymph nodes occurs in three distinct phases. Nature 427:154–159CrossRefPubMedGoogle Scholar
  34. Mempel TR, Scimone ML, Mora JR, von Andrian UH (2004b) In vivo imaging of leukocyte trafficking in blood vessels and tissues. Curr Opin Immunol 16:406–417CrossRefGoogle Scholar
  35. Miller MJ, Wei SH, Parker I, Cahalan MD (2002) Two-photon imaging of lymphocyte motility and antigen response in intact lymph node. Science 296:1869–1873CrossRefPubMedGoogle Scholar
  36. Niewöhner J, Weber I, Maniak M, Müller-Tauchenberger A, Gerisch G (1997) Talin-null cells from Dictyostelium are strongly defective in adhesion to particle and substrate surfaces and slightly impaired in cytokinesis. J Cell Biol 138:349–361CrossRefPubMedGoogle Scholar
  37. Petroll WM, Ma L (2003) Direct, dynamic assessment of cell-matrix interactions inside fibrillar collagen lattices. Cell Motil Cytoskeleton 55:254–264CrossRefPubMedGoogle Scholar
  38. Petroll WM, Cavanagh HD, Jester JV (2004) Dynamic three-dimensional visualization of collagen matrix remodeling and cytoskeletal organization in living corneal fibroblasts. Scanning 26:1–10PubMedGoogle Scholar
  39. Ploem JS (1975) Reflection-contrast microscopy as a tool for investigation of the attachment of living cells to a glass surface. In: van Furth R (ed) Handbook of biological confocal microscopy. Blackwell, Oxford, pp 404–421Google Scholar
  40. Rieckhoff KE, Peticolas WL (1965) Optical second-harmonic generation in crystalline amino acids. Science 147:610–611PubMedGoogle Scholar
  41. Rosso F, Giordano A, Barbarisi M, Barbarisi A (2004) From cell-ECM interactions to tissue engineering. J Cell Physiol 199:174–180CrossRefPubMedGoogle Scholar
  42. Seftor RE, Seftor EA, Koshikawa N, Meltzer PS, Gardner LM, Bilban M, Stetler-Stevenson WG, Quaranta V, Hendrix MJ (2001) Cooperative interactions of laminin 5 gamma2 chain, matrix metalloproteinase-2, and membrane type-1-matrix/metalloproteinase are required for mimicry of embryonic vasculogenesis by aggressive melanoma. Cancer Res 61:6322–6327PubMedGoogle Scholar
  43. Simian M, Hirai Y, Navre M, Werb Z, Lochter A, Bissell MJ (2001) The interplay of matrix metalloproteinases, morphogens and growth factors is necessary for branching of mammary epithelial cells. Development 128:3117–3131PubMedGoogle Scholar
  44. Tamariz E, Grinnell F (2002) Modulation of fibroblast morphology and adhesion during collagen matrix remodeling. Mol Biol Cell 13:3915–3929CrossRefPubMedGoogle Scholar
  45. Vanni S, Lagerholm BC, Otey C, Taylor DL, Lanni F (2003) Internet-based image analysis quantifies contractile behavior of individual fibroblasts inside model tissue. Biophys J 84:2715–2727PubMedGoogle Scholar
  46. Voytik-Harbin SL, Roeder BA, Sturgis JE, Kokini K, Robinson JP (2003) Simultaneous mechanical loading and confocal reflection microscopy for three-dimensional microbiomechanical analysis of biomaterials and tissue constructs. Microsc Microanal 9:74–85CrossRefPubMedGoogle Scholar
  47. Wang W, Wyckoff JB, Frohlich VC, Oleynikov Y, Huttelmaier S, Zavadil J, Cermak L, Bottinger EP, Singer RH, White JG, Segall JE, Condeelis JS (2002) Single cell behavior in metastatic primary mammary tumors correlated with gene expression patterns revealed by molecular profiling. Cancer Res 62:6278–6288PubMedGoogle Scholar
  48. Webb DJ, Brown CM, Horwitz AF (2003) Illuminating adhesion complexes in migrating cells: moving toward a bright future. Curr Opin Cell Biol 15:614–620CrossRefPubMedGoogle Scholar
  49. Wolf K, Mazo I, Leung H, Engelke K, von Andrian UH, Deryugina EI, Strongin AY, Brocker EB, Friedl P (2003a) Compensation mechanism in tumor cell migration: mesenchymal-amoeboid transition after blocking of pericellular proteolysis. J Cell Biol 160:267–277CrossRefPubMedGoogle Scholar
  50. Wolf K, Muller R, Borgmann S, Brocker EB, Friedl P (2003b) Amoeboid shape change and contact guidance: T-lymphocyte crawling through fibrillar collagen is independent of matrix remodeling by MMPs and other proteases. Blood 102:3262–3269CrossRefPubMedGoogle Scholar
  51. Yang Y, Whiteman S, van Pittius DG, He Y, Wang RK, Spiteri MA (2004) Use of optical coherence tomography in delineating airways microstructure: comparison of OCT images to histopathological sections. Phys Med Biol 49:1247–1255CrossRefPubMedGoogle Scholar
  52. Yeh AT, Choi B, Nelson JS, Tromberg BJ (2003) Reversible dissociation of collagen in tissues. J Invest Dermatol 121:1332–1335PubMedGoogle Scholar
  53. Yeh AT, Kao B, Jung WG, Chen Z, Nelson JS, Tromberg BJ (2004) Imaging wound healing using optical coherence tomography and multiphoton microscopy in an in vitro skin-equivalent tissue model. J Biomed Opt 9:248–253CrossRefPubMedGoogle Scholar
  54. Zipfel WR, Williams RM, Christie R, Nikitin AY, Hyman BT, Webb WW (2003) Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation. Proc Natl Acad Sci U S A 100:7075–7080CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Rudolf-Virchow Center, DFG Research Center for Experimental Biomedicine, and Department of DermatologyUniversity of WürzburgWürzburgGermany

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