Abstract
The primary aim of tissue fixation is to preserve tissue in the long term, ideally without causing alterations in morphology or biochemical integrity. Optimal fixation should therefore inhibit autolysis while preserving enzyme activity and antigen reactivity. However, some of these requirements are mutually exclusive. For example, to stop autolysis, proteolytic enzymes must be inhibited (1). At present, it is not possible to find a fixation technique that fulfils all of these criteria.
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Asplund, M. and Brunk, U. (1975) Osmotiska forhallen under aldehydfixering. Laboratoriet 9, 1–11.
Fox, C. H., Johnson, F. B., Whiting, J., and Roller P. P. (1985) Formaldehyde fixation. J. Histochem. Cytochem. 33, 845–853.
Bancroft, J. D. and Stevans, A. (1999) Theory & Practice of Histological Techniques 4th ed, Churchill Livingstone, New York.
Walker, J. F. (1964) Formaldehyde, in American Chemical Society Monograph Series, 3rd ed., Reinhold, New York.
Feldman, M. Y. (1973) Reactions of nucleic acids and nucleoproteins with formaldehyde. Prog. Nucleic Acid Res. Mol. Biol. 13, 1–49.
Ruijter, E. T., Miller, G., Aalders, T. W., Van De Kaa, C. A., Schalken, J. A., Debruyne, F. M., et al. (1997) Rapid microwave-stimulated fixation of entire prostatectomy specimens J. Pathol. 183, 369–375.
Baak, J. P. A. (1991) Manual of Quantitative Pathology in Cancer Diagnosis and Prognosis, Springer-Verlag, Berlin and Heidelberg.
Taylor, C. R., Shi, S. R., and Cote, R. J. (1996) Antigen retrieval for immunohistochemistry. Status and need for greater standardization. Appl. Immunohistochem. 4, 144–166.
Shi, S.-R., Imam A., Young, L., Cote, R. J., and Taylor C. R. (1995) Antigen retrieval immunohistochemistry under influence of pH using monoclonal antibodies J. Histochem. Cytochem. 43, 193–201.
Shi, S.-R., Cote, R. J., and Taylor C. R. (1997) Antigen retrieval immunohisto-chemistry: Past, present and the future. J. Histochem. Cytochem. 45, 327–343.
Shi, S.-R., Cote, R. J., Young, L. L., and Taylor, C. R. (1997) Antigen retrieval immunohistochemistry: Practice and development. J. Histotechnology 20, 145–154.
Pileri, S. A., Roncandor, G., Ceccarelli, C., Piccioli, M., Briskomatis, A., Sabbatini, E., et al. (1997) Antigen retrieval techniques in immunohistochemistry: Comparison of different methods. J. Pathol. 183, 116–123.
Leong, A. S. Y. and Gilham, P. N. (1989) The effects of progressive formaldehyde fixation on the preservation of tissue antigens. Pathology 21, 266–268.
Login, G. R., Schnitt, S. J., and Dovrak, A. M. (1987) Rapid microwave fixation of human tissues for light microscopic immunoperoxidase identification of diagnostically useful antigens. Lab. Invest. 57, 585–591.
Cohen, R. J., McNeal, J. E., Meehan, K., and Wilce, M. (2000) Luminal contents of benign and malignant prostatic glands: Correspondence to altered secretory mechanisms. Hum. Pathol. 32, 94–101.
Cohen, R. J. and Beales, M. (2000) Prostate secretory granules in normal and neoplastic prostate glands: A diagnostic aid to needle biopsy. Hum. Pathol. 31, 1515–1519.
Cohen, R. J., Fischer, G., and McNeal, J. E. (2001) Immunostains after gluteraldehyde-based tissue fixation. Hum. Pathol. 32, 242–243.
Arnold, M. M., Srivastava, S., Fredenburgh, J., Stockard, C. R., Myers, R. B., and Grizzle, W. E. (1996) Effects of fixation and tissue processing on immunohistochemical demonstration of specific antigens. Biotech. Histochem. 71, 224–230.
Gillespie, J. W., Best, C. J. M., Bichel, V. E., Cole, K. A., Greenhut, S. F., Hewitt, S. M., et al. (2002) Evaluation of non-formalin tissue fixation for molecular profiling studies. Am. J. Pathol. 160, 449–457.
Russell, P. and Farnsworth, A. (1997) Examination of the ovarian specimen, in Surgical Pathology of the Ovaries (Russell, P. and Farnsworth, A., eds.), Churchill Livingstone, Edinburgh, pp. 25–37.
Goelz, S. E., Hamilton, S. R., and Vogelstein, B. (1985) Purification of DNA from formaldehyde fixed and paraffin embedded human tissue. Biochem. Biophys. Res. Commun. 130, 118–126.
Dubeau L., Chandler, L. A., Gralow, J. R., Nichols, P. W., and Jones, P. A. (1986) Southern blot analysis of DNA extracted from formalin-fixed, paraffin-embedded pathology specimens. Cancer Res. 46, 2964–2969.
Wong, A. J., Ruppert, J. M., Eggelston, J., Hamilton, S. R., Baylin, S. B., and Vogelstein, B. (1986) Gene amplification of c-myc and N-myc in small cell carcinoma of the lung. Science 233, 461–464.
Tsuda, H., Shimosato, Y., Upton, M. P., Yokota, J., Tereda, M., Okira, M., et al. (1988) Retrospective study on amplification of N-myc and c-myc genes in pediatric solid tumors and its association with prognosis and tumor differentiation. Lab. Invest. 59, 321–327.
Mies, C., Houldsworth, J., and Changanti, R. S. K. (1991) Extraction of DNA from paraffin blocks for Southern blot analysis. Am. J. Surg. Pathol. 15, 169–174.
Park, J. S., Leake, J. F., Sharma, B. K., Toki, T., Kessis, T. O., Ambros, R. A., et al. (1991) Variability in beta-globin and HPV DNA amplification by PCR from fixed tissues. Mod. Pathol. 4, 667–670.
Honma, M., Ohara, Y., Murayama, H., Saho, K., and Ivasali, Y. (1993) Effects of fixation and varying target length on the sensitivity of polymerase chain reaction for detection of human T-cell leukemia virus type I proviral DNA in formalin-fixed tissue sections. J. Clin. Microbiol. 31, 1799–1803.
Stanta, G. and Schneider, C. (1991) RNA extracted from paraffin-embedded human tissue is amenable to analysis of PCR amplification. Biotechniques 11, 304–408.
Woodall, C. J., Watt, N. J., and Clements, G. B. (1993) Simple technique for detecting RNA viruses by PCR in single sections of wax embedded tissue. J. Clin. Pathol. 46, 276–277.
Hilton, D. A., Variend, S., and Pringle, J. H. (1993) Demonstration of coxackie virus RNA in formalin-fixed tissue sections from childhood myocarditis cases by in situ hybridization and the polymerase chain reaction. J. Pathol. 170, 45–51.
Akyol, G., Dash, S., Shieh, Y. S., Malter, J. S., and Gerber, M. A. (1992) Detection of hepatitis C virus RNA sequences by polymerase chain reaction in fixed live tissue. Mod. Pathol. 5, 501–504.
Jackson, D. P., Quirke, P., Lewis, F., Boylston, A.W., Sloan, J. M., Robertson, D., and Taylor, J. R. (1989) Detection of measles virus RNA in paraffin-embedded tissue (letter). Lancet 1, 1391.
Redline, R. W., Genest, D., and Tycko B. (1991) Detection of enteroviral infection in paraffin-embedded tissue by the RNA polymerase chain reaction technique. Am. J. Clin. Pathol. 96, 568–571.
Navas, S., Castillo, I., and Carreno, V. (1993) Detection of plus and minus HCV RNA in normal liver of anti-HCV-positive patients (Letter). Lancet 341, 904–905.
Bramwell, N. H. and Burns B. F. (1988) The effect of fixative type and fixation time on the quantity and quality of extractable DNA for hybridization studies on lymphoid tissue. Exp. Hematol. 16, 730–732.
Rogatch, H., Moser, P., Volgger, H., Horninger, W., Bartsch, G., Mikuz, G., et al. (2000) Diagnostic effect of an improved pre-embedding method of prostate needle biopsy specimen. Hum. Pathol. 31, 1102–1107.
Egevad, L., Engstrom, K., and Busch, C. (1998) A new method for handling radical prostatectomies enabling fresh tissue harvesting, whole mount sections, and landmarks for alignment of sections. J. Urol. Pathol. 9, 17–26.
Church, R. J., Hand, N. M., Rex, M., and Scotting, P. J. (1997) Non-isotopic in situ hybridisation to detect Sox gene mRNA in plastic-embedded tissue. Histochem. J. 29, 625–629.
Warren, K. C., Coyne, K. J., Waite, J. H., and Cary, S. C. (1998) Use of methacrylate de-embedding protocols for in situ hybridisation on semithin plastic sections with multiple detection strategies. J. Histochem. Cytochem. 46, 149–155.
McCluggage, W. G., Roddy, S., Whiteside, C., Burton, J., McBride, H., Maxwell, P., et al. (1995) Immunohistochemical staining of plastic embedded bone marrow trephine biopsy specimens after microwave heating. J. Clin. Pathol. 48, 840–844.
Brown, R. S., Edwards, J., Bartlett, J. W., Jones, C., and Dogan, A. (2002) Routine acid decalcification of bone marrow samples can preserve DNA for FISH and CGH studies in metastatic prostate cancer. J. Histochem. Cytochem. 5, 113–116.
Alers, J. C., Krijtenburg, P.-J., Vissers, K. J., and van Dekken, H. (1999) Effect of bone decalcification procedures on DNA in situ hybridisation and comparative genomic hybridisation: EDTA is highly preferable to a routinely used acid decalcifier. J. Histochem. Cytochem. 47, 703–709.
Lebeau, A., Muthmann, H., Sendelhofert, A., Diebold, J., and Lohrs, U. (1995) Histochemistry and immunohistochemistry on bone marrow biopsies. A rapid procedure for methyl methacrylate embedding. Pathol. Res. Practice 191, 121–129.
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Mortensen, E., Brown, J.M. (2003). Effects of Fixation on Tissues. In: Russell, P.J., Jackson, P., Kingsley, E.A. (eds) Prostate Cancer Methods and Protocols. Methods in Molecular Medicine™, vol 81. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-372-0:163
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DOI: https://doi.org/10.1385/1-59259-372-0:163
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-978-0
Online ISBN: 978-1-59259-372-9
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