Skip to main content
SpringerLink
Log in

The effect of different fixatives and length of fixation time on subsequent AgNOR staining for frozen and paraffin-embedded tissue sections

  • Papers
  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Summary

The AgNOR technique has been used extensively in studies investigating the possibility that the numbers and appearances of the intranuclear structures stained are markers of malignancy. The method has the advantage of being applicable to many different types of histological material, including paraffin-embedded tissue. However, it has been suggested that the visualization of AgNORs is dependent on the type and time of fixation employed. This study set out to measure this effect with the following commonly-used fixatives: acetone, absolute ethanol, methanol, Carnoy's fluid, Bouin's fluid, 4% glutaraldehyde, 10% neutral buffered formalin and 10% formol-saline. Both frozen sections and blocks of fresh tonsil were fixed for varying times, the blocks of tissue then being processed routinely. With the frozen sections AgNORs were easier to discern than in sections of paraffin-embedded tissue, and more intranucleolar AgNORs were visible when alcoholic fixatives were used than with aldehyde fixation. The effects of different fixatives on AgNOR appearance in paraffin sections is, however, more complex. Despite the variation caused by different fixatives, AgNORs could be demonstrated adequately with all the fixatives studied. It is concluded that fixation is not a limitation to the study of AgNORs provided that the time and type of fixative is controlled.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bancroft, J. D. & Stevens, A. (1982) Theory and Practice of Histological Techniques, 2nd edn. Edinburgh: Churchill Livingstone.

    Google Scholar 

  • Chui, K. Y., Loke, S. L. & Wong, K. K. (1989) Improved silver technique for showing nucleolar organizer regions in paraffin wax sections. J. Clin. Pathol. 42, 922–4.

    Google Scholar 

  • Crocker, J. (1990) Nucleolar organizer regions. In Current Topics in Pathology, Vol 82, Pathology of the Nucleus (edited by Underwood, J. C. E.) pp. 91–149. Berlin: SpringerVerlag.

    Google Scholar 

  • Crocker, J., Boldy, D. A. R. & Egan, M. J. (1989) How should we count AgNORs? Proposals for a standardized approach. J. Pathol. 158, 185–8.

    Google Scholar 

  • Cromie, C. J., Benbow, E. W., Stoddart, R. W. & Mcmahon, R. F. T. (1988) Preincubation with a glycine solution aids the demonstration of nucleolar organizer region-associated protein. Histochem. J. 20, 722–4.

    Google Scholar 

  • Derenzini, M., Romagnoli, T., Ceccarelli, C. & Eusibi, V. (1988) Fixatives and silver stainability of NOR proteins at the light microscopic level. J. Histochem. Cytochem. 36, 1453–4.

    Google Scholar 

  • Griffiths, A. P., Butler, C. W., Roberts, P., Dixon, M. F. & Quirke, P. (1989) Silver-stained structures (AgNORs), their dependence on tissue fixation and absence of prognostic relevance in rectal adenocarcinoma. J. Pathol. 159, 121–7.

    Google Scholar 

  • Leong, A. S-Y. & Raymond, W. (1988) Demonstration of AgNOR-related proteins in microwave-fixed tissues. J. Pathol. 156, 352.

    Google Scholar 

  • Ploton, D., Menager, M., Jeannesson, P., Himber, G., Pigeon, F. & Adnet, J. J. (1986) Improvement in the staining and in the visualization of the argyrophilic proteins of the nucleolar organizer region at the optical level. Histochem. J. 18, 5–14.

    Google Scholar 

  • Rowlands, D. C., Crocker, J. & Ayres, J. G. (1990) Silver staining of nucleolar organizer region associated proteins using polyethylene glycol as the protective colloidal developer. Histochem. J. 22, 555–9.

    Google Scholar 

  • Schwarzacher, H. G. & Wachtler, F. (1983) Nucleolus organizer regions and nucleoli. Human Genetics 63, 89–99.

    Google Scholar 

  • Smith, R. & Crocker, J. (1988) Evaluation of nucleolar organizer region-associated proteins in breast malignancy. Histopathology 12, 113–25.

    Google Scholar 

  • Smith, P. J., Skilbeck, N., Harrison, A. & Crocker, J. (1988) The effect of a series of fixatives on the AgNOR technique. J. Pathol. 155, 109–12.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Histopathology, East Birmingham Hospital, B9 5ST, Birmingham, West Midlands, UK

    David C. Rowlands & John Crocker

  2. Department of Respiratory Medicine, East Birmingham Hospital, B9 5ST, Birmingham, West Midlands, UK

    Jon G. Ayres

Authors
  1. David C. Rowlands
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jon G. Ayres
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. John Crocker
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rowlands, D.C., Ayres, J.G. & Crocker, J. The effect of different fixatives and length of fixation time on subsequent AgNOR staining for frozen and paraffin-embedded tissue sections. Histochem J 25, 123–132 (1993). https://doi.org/10.1007/BF00157984

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00157984

Keywords

Search

Navigation