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Evaluation of non-formalin tissue fixation for molecular profiling studies

  1. Author:
    Gillespie, J. W.
    Best, C. J. M.
    Bichsel, V. E.
    Cole, K. A.
    Greenhut, S. F.
    Hewitt, S. M.
    Ahram, M.
    Gathright, Y. B.
    Merino, M. J.
    Strausberg, R. L.
    Epstein, J. I.
    Hamilton, S. R.
    Gannot, G.
    Baibakova, G. V.
    Calvert, V. S.
    Flaig, M. J.
    Chuaqui, R. F.
    Herring, J. C.
    Pfeifer, J.
    Petricoin, E. F.
    Linehan, W. M.
    Duray, P. H.
    Bova, G. S.
    Emmert-Buck, M. R.

  2. Author Address

    NCI, Pathol Lab, Pathogenet Unit, Rm 2A33, Bldg 10, 9000 Rockville Pike, Bethesda, MD 20892 USA. NCI, Pathol Lab, Pathogenet Unit, Bethesda, MD 20892 USA. NCI, Sci Applicat Int Corp, Frederick, MD 21701 USA. US FDA, Ctr Biolog & Res, Bethesda, MD 20014 USA. NCI, Off Director, CGAP, Bethesda, MD 20892 USA. Johns Hopkins Univ, Dept Pathol, Baltimore, MD USA. Univ Texas, MD Anderson Canc Ctr, Dept Pathol, Houston, TX 77030 USA. Tel Aviv Univ, Sackler Fac Med, IL-69978 Tel Aviv, Israel. Ctr Prostate Dis Res, Rockville, MD USA. NIH, Ctr Informat Technol, Bethesda, MD 20892 USA. NCI, Urol Oncol Branch, Bethesda, MD 20892 USA. Emmert-Buck MR NCI, Pathol Lab, Pathogenet Unit, Rm 2A33, Bldg 10, 9000 Rockville Pike, Bethesda, MD 20892 USA.
    1. Year: 2002
  2. Journal: American Journal of Pathology
    1. 160
    2. 2
    3. Pages: 449-457
  4. Type of Article: Article
  1. Abstract:

    Using a general strategy for evaluating clinical tissue specimens, we found that 70% ethanol fixation and paraffin embedding is a useful method for molecular profiling studies. Human prostate and kidney were used as test tissues. The protein content of the samples was analyzed by one-dimensional gel electrophoresis, immunoblot, two-dimensional gel electrophoresis, and layered expression scanning. In each case, the fixed and embedded tissues produced results similar to that obtained from snap-frozen specimens, although the protein quantity was somewhat decreased. Recovery of mRNA was reduced in both quantity and quality in the ethanol-fixed samples, but was superior to that obtained from formalin-fixed samples and sufficient to perform reverse transcription polymerase chain reactions. Recovery of DNA from ethanol-fixed specimens was superior to formalin-fixed samples as determined by one- dimensional gel electrophoresis and polymerase chain reaction. In conclusion, specimens fixed in 70% ethanol and embedded in paraffin produce good histology and permit recovery of DNA, mRNA, and proteins sufficient for several downstream molecular analyses. Complete protocols and additional discussion of relevant issues are available on an accompanying website (http://cgap-mf.nih.gov/).

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