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. 1968 Jan 1;36(1):231–243.

THE USE OF MILLIPORE FILTERS IN ULTRASTRUCTURAL STUDIES OF CELL CULTURES AND VIRUSES

Robert M McCombs 1, Matilda Benyesh-Melnick 1, J Pierre Brunschwig 1
PMCID: PMC2107342  PMID: 19806703

Abstract

A technique is described for embedding tissue culture cells that have been adsorbed or grown on Millipore filters. The acetone used during the embedding process rendered the filters transparent so that specific areas or cells could be chosen with the aid of the light microscope. Lymphoblastoid cells processed on the filters possessed well-defined plasma membranes and microvilli which were rarely present in cells from parallel cultures that were prepared by pelleting in the centrifuge. Fibroblast cells grown on filters retained their elongated appearance, in contrast to the rounded cells in pelleted preparations. Millipore filters were also used as a means of embedding virus pellets for sectioning. Preparations containing as few as 4 x 108 virus particles were suitable for study by the filter technique. Crude tissue-culture harvests of vaccinia virus and purified preparations of Rauscher murine leukemia and adeno-satellite viruses were successfully examined.

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Selected References

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

  1. BENYESH-MELNICK M., FERNBACH D. J., LEWIS R. T. STUDIES ON HUMAN LEUKEMIA. I. SPONTANEOUS LYMPHOBLASTOID TRANSFORMATION OF FIBROBLASTIC BONE MARROW CULTURES DERIVED FROM LEUKEMIC AND NONLEUKEMIC CHILDREN. J Natl Cancer Inst. 1963 Dec;31:1311–1331. [PubMed] [Google Scholar]
  2. BLOOM W. Preparation of a selected cell for electron microscopy. J Biophys Biochem Cytol. 1960 Feb;7:191–194. doi: 10.1083/jcb.7.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BORYSKO E. Recent developments in methacrylate embedding. II. Methods for the sectioning of optically selected single cells, the orientation of the plane of sectioning and the identification of the region of the specimen included in the sections. J Biophys Biochem Cytol. 1956 Jul 25;2(4 Suppl):15–20. doi: 10.1083/jcb.2.4.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. BORYSKO E., SAPRANAUSKAS P. A new technique for comparative phase-contrast and electron microscope studies of cells grown in tissue culture, with an evaluation of the technique by means of time-lapse cinemicrographs. Bull Johns Hopkins Hosp. 1954 Aug;95(2):68–79. [PubMed] [Google Scholar]
  5. Baudhuin P., Evrard P., Berthet J. Electron microscopic examination of subcellular fractions. I. The preparation of representative samples from suspensions of particles. J Cell Biol. 1967 Jan;32(1):181–191. doi: 10.1083/jcb.32.1.181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. GAY H. Serial sections of smears for electron microscopy. Stain Technol. 1955 Sep;30(5):239–242. doi: 10.3109/10520295509114472. [DOI] [PubMed] [Google Scholar]
  7. GROBSTEIN C. Trans-filter induction of tubules in mouse metanephrogenic mesenchyme. Exp Cell Res. 1956 Apr;10(2):424–440. doi: 10.1016/0014-4827(56)90016-7. [DOI] [PubMed] [Google Scholar]
  8. HOWATSON A. F., ALMEIDA J. D. A method for the study of cultured cells by thin sectioning and electron microscopy. J Biophys Biochem Cytol. 1958 Jan 25;4(1):115–118. doi: 10.1083/jcb.4.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. LATTA H. A cellular reaction to antibody in tissue culture studied with electron microscopy. J Biophys Biochem Cytol. 1959 May 25;5(3):405–410. doi: 10.1083/jcb.5.3.405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. MINICK O. T., NEBEL B. R. Embedding aid for coverslip mounts in electron microscopy. J Biophys Biochem Cytol. 1956 Jul 25;2(4 Suppl):61–63. doi: 10.1083/jcb.2.4.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. McCombs R. M., Melnick M. B., Brunschwig J. P. Biophysical studies of vesicular stomatitis virus. J Bacteriol. 1966 Feb;91(2):803–812. doi: 10.1128/jb.91.2.803-812.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. NISHIURA M., RANGAN S. R. A device for embedding tissue culture preparations grown on coverslips. J Biophys Biochem Cytol. 1960 Apr;7:411–412. doi: 10.1083/jcb.7.2.411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. O'CONNOR T. E., RAUSCHER F. J., ZEIGEL R. F. DENSITY GRADIENT CENTRIFUGATION OF A MURINE LEUKEMIA VIRUS. Science. 1964 May 29;144(3622):1144–1147. doi: 10.1126/science.144.3622.1144. [DOI] [PubMed] [Google Scholar]
  14. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. ROSEN S. I. Cover-glass embedding in open-end capsules for electron microscopy. Stain Technol. 1962 May;37:195–197. doi: 10.3109/10520296209117735. [DOI] [PubMed] [Google Scholar]
  16. Rabin E. R., Benyesh-Melnick M., Brunschwig J. P. Studies on acute leukemia and infectious mononucleosis of childhood. II. Ultrastructure of cultured lymphoblastoid cells. Exp Mol Pathol. 1967 Oct;7(2):196–207. doi: 10.1016/0014-4800(67)90029-9. [DOI] [PubMed] [Google Scholar]
  17. Ross R., Lillywhite J. W. The fate of buffy coat cells grown in subcutaneously implanted diffusion chambers. A light and electron microscopic study. Lab Invest. 1965 Sep;14(9):1568–1585. [PubMed] [Google Scholar]
  18. SHAFFER B. M. The culture of organs from the embryonic chick on cellulose acetate fabric. Exp Cell Res. 1956 Aug;11(1):244–248. doi: 10.1016/0014-4827(56)90217-8. [DOI] [PubMed] [Google Scholar]

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