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A comparison of methods for morphological studies of cultured cells

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Summary

A number of fixation methods for different types of cells in culture were compared, and the best preservation of nuclear and cytoplasmic details was obtained by fixation with Bouin's solution for 15 min, prior to staining with hematoxylin and eosin. All of the fixatives, including Bouin's solution, damaged various structures, notably the peripheral glas-attached cytoplasm and the intercellular connections. Micrographs obtained by bright field, phase contrast, and interference contrast (Nomarski) microscopy are presented. Much more realistic pictures, bringing out details not observed after fixation and staining, were obtained by Nomarski microscopy of living, unfixed cultures. Most conspicuous were numerous thin, cytoplasmic, cilia-like extensions, concentrated on the glass-attached peripheral margins, which were also visible on other cell surfaces and as intercellular connections. These structures were most characteristic of SV40-transformed human amnion cells. Although fixation and staining emphasize certain cell components (for example, inclusion bodies), many aspects of cellular morphology are better demonstrated by observing living cells by interference microscopy or by Nomarski interference contrast microscopy. Surface features of unfixed cells, seen by Nomarski interference contrast microscopy, were similar to the surface features of glutaraldehyde-osmium tetroxide-fixed cells studied as metallic replicas in the electron microscope.

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References

  1. Sykes, J. A., J. Whitescarver, L. Briggs, and J. H. Anson. 1970. Separation of tumor cells from fibroblasts with use of discontinuous density gradients. J. Natl. Cancer Inst. 44: 855–864.

    PubMed  CAS  Google Scholar 

  2. Fogh, J., L. Ramos, and H. Fogh. 1969. Transformation of primary human amnion cells with simian virus 40. In: G. L. Tritsch (Ed.),Axenic Mammalian Cell Reactions. Marcel Dekker. New York, pp. 59–116.

    Google Scholar 

  3. Gaffney, E. V., J. Fogh, L. Ramos, J. Loveless, H. Fogh, and A. M. Dowling. 1970. Established lines of SV40 transformed human amnion cells. Cancer Res. 30: 1668–1676.

    PubMed  CAS  Google Scholar 

  4. McCoy, T., M. Maxwell, and P. Kruse. 1959. Amino acid requirements of the Novikoff hepatomain vitro. Proc. Soc. Exp. Biol. Med. 100: 115–118.

    PubMed  CAS  Google Scholar 

  5. Hayflick, L. 1965. The limitedin vitro lifetime of human diploid cell strains. Exp. Cell Res. 37: 614–636.

    Article  PubMed  CAS  Google Scholar 

  6. Fogh, J., and R. O. Lund. 1957. Continuous cultivation of epithelial cell strain (FL) from human amniotic membrane. Proc. Soc. Exp. Biol. Med. 94: 532–537.

    PubMed  CAS  Google Scholar 

  7. Leighton, J. 1951. A sponge matrix method for tissue culture. Formation of organized aggregates of cells in vitro. J. Natl. Cancer Inst. 12: 545–562.

    PubMed  CAS  Google Scholar 

  8. Sykes, J. A., J. Whitescarver, P. Jernstrom, and J. F. Nolan. 1970. Some properties of a new epithelial cell line of human origin. J. Natl. Cancer Inst. 45: 107–122.

    PubMed  CAS  Google Scholar 

  9. Sykes, J. A., and E. B. Moore 1960. A simple tissue culture chamber. Texas Rep. Biol. Med. 18: 288–297.

    CAS  Google Scholar 

  10. Fogh, J., H. Fogh, and L. Ramos. 1971. Growthin vitro of mycoplasma-infected human amnion cells, FL amnion cells, and mycoplasma-modified FL cells. Proc. Soc. Exp. Biol. Med. 136: 809–818.

    PubMed  CAS  Google Scholar 

  11. Nomarski, G. 1955. Microinterférométrie différentielle à ondes polarisées. J. Physique Radium 16: 9–13.

    Google Scholar 

  12. Hale, A. J. 1958.The Interference Microscope in Biological Research, E. & S. Livingstone, Edinburgh and London.

    Google Scholar 

  13. Hanks, J. H., and R. E. Wallace. 1949. Relation of oxygen and temperature in the preservation of tissues for refrigeration. Proc. Soc. Exp. Biol. Med. 71: 196–200.

    Google Scholar 

  14. Millonig, G. 1961. Advantages of a phosphate buffer for OsO4 solutions in fixation. Proc. 19th Annual Meeting Electron Microscopy Society of America, pp. 15–16.

  15. Coman, D. R., and T. F. Anderson. 1955. A structural difference between the surfaces of normal and of carcinomatous epidermal cells. Cancer Res. 15: 541–544.

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Sloan-Kettering Institute for Cancer Research, 10021, New York, New York

    Jørgen Fogh & John A. Sykes

  2. Research Department, Southern California Cancer Center, 90015, Los Angeles, California

    Jørgen Fogh & John A. Sykes

Authors
  1. Jørgen Fogh
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  2. John A. Sykes
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Supported in part by National Cancer Institute Research Grant CA-08748 and contributions from the Albert Soiland Cancer Foundation.

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Fogh, J., Sykes, J.A. A comparison of methods for morphological studies of cultured cells. In Vitro 7, 206–227 (1972). https://doi.org/10.1007/BF02615977

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