Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Bio-Technical Methods Section (BTS)

Simultaneous detection of the immunophenotypic markers and genetic aberrations on routinely processed paraffin sections of lymphoma samples by means of the FICTION technique


Disciplines such as morphology, immunophenotyping and genetics widely contributed over decades to the understanding of the cellular mechanisms of cancer. To obtain a greater insight into the complex processes of tumorigenesis, scientists have joined their efforts to combine many of the available techniques. Here, we report on the development of a FICTION (Fluorescence Immunophenotyping and Interphase Cytogenetics as a tool for the Investigation of Neoplasms) technique that allows a simultaneous detection of immunophenotypic markers and genetic aberrations on routinely processed lymphoma samples. As the antigen retrieval method seems to play an important role in the final results, we tested the pressure-cooking method at different times (2, 4 and 8 min) using three different buffers (EDTA, Tris-EDTA and citrate), resulting in improved sensitivity for the detection of both immunophenotypic markers and genetic aberrations. We also applied this method to different types of lymphoma using double immunofluorescence assays (including CD30, CD20, CD8 monoclonal antibodies) and several fluorescence in situ hybridization probes to demonstrate that the FICTION technique could be easily applied on paraffin sections in different combinations for the diagnosis and research of cancer.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1
Figure 2


  1. Weber-Matthiesen K, Winkemann M, Muller-Hermeling A, Schlegelberger B, Grote W . Simultaneous fluorescence immunophenotyping and interphase cytogenetics: a contribution to the characterization of tumour cells. J Histochem Cytochem 1992; 40: 171–175.

    Article  CAS  Google Scholar 

  2. Weber-Matthiesen K, Muller-Hermelink A, Deerberg J, Scherthan H, Schlegelberger B, Grote W . Discrimination of distinct subpopulations within a tumour with combined double immunophenotyping and interphase cytogenetics. J Histochem Cytochem 1993; 41: 1641–1644.

    Article  CAS  Google Scholar 

  3. Weber-Matthiesen K, Deerberg J, Poetsch M, Grote W, Schlegelberger B . Rapid immunophenotypic characterization of chromosomally aberrant cells by the new FICTION method. Cytogenet Cell Genet 1993; 63: 123–125.

    Article  CAS  Google Scholar 

  4. Weber-Matthiesen K, Pressl S, Schlegelberger B, Grote W . Combined immunophenotyping and interphase cytogenetics on cryostat sections by the new FICTION method. Leukemia 1993; 7: 646–649.

    CAS  PubMed  Google Scholar 

  5. Zandecki M, La JL, Genevieve F, Bernardi F, Volle R, Blanchet O et al. Several cytogenetic subclones may be identified within plasma cells from patients with monoclonal gammopathy of undetermined significance, both at diagnosis and during the indolent course of this condition. Blood 1997; 9: 3682–3690.

    Google Scholar 

  6. Gesk S, Martin-Subero JI, Harder L, Luhmann B, Schlegelberger B, Calasanz MJ et al. Molecular cytogenetic detection of chromosomal breakpoints in T-cell receptor gene loci. Leukemia 2003; 17: 738–745.

    Article  CAS  Google Scholar 

  7. Barth TFE, Martin-Subero JI, Joos S, Menz CK, Hasel C, Mechtersheimer G et al. Gains of 2p involving the REL locus correlate with nuclear c-Rel protein accumulation in neoplastic cells of classical Hodgkin lymphoma. Blood 2003; 101: 3681–3686.

    Article  CAS  Google Scholar 

  8. Martin-Subero JI, Chudoba I, Harder L, Gesk S, Grote W, Novo FJ et al. Multicolor-FICTION. Am J Pathol 2002; 161: 413–420.

    Article  CAS  Google Scholar 

  9. Haralambieva E, Banham AH, Bastard C, Delsol G, Gaulard P, Ott G et al. Detection by the fluorescence in situ hybridization technique of MYC translocations in paraffin-embedded lymphoma biopsy samples. Br J Haematol 2003; 121: 49–56.

    Article  CAS  Google Scholar 

  10. Haralambieva E, Kleiverda K, Mason DY, Schuuring E, Kluin PM . Detection of three common translocation breakpoints in non-Hodgkin's lymphomas by fluorescence in situ hybridization on routine paraffin-embedded tissue sections. J Pathol 2002; 198: 163–170.

    Article  CAS  Google Scholar 

  11. Moreno-Bueno G, Rodríguez-Perales S, Sánchez-Estévez C, Hardisson D, Sarrió D, Prat J, Cigudosa JC et al. CyclinD1 gene (CCND1) mutations in endometrial cancer. Oncogene 2003; 4: 6115–6118.

    Article  Google Scholar 

  12. Pileri AS, Roncador G, Ceccarelli C, Piccioli M, Briskomatis A, Sabattini E et al. Antigen retrieval techniques in immunohistochemistry: comparison of different methods. J Pathol 1997; 183: 116–123.

    Article  CAS  Google Scholar 

  13. Morgan JM, Navabi H, Schmid KW, Jasani B . Possible role of tissue-bound calcium ions in citrate-mediated high-temperature antigen retrieval. J Pathol 1994; 174: 301–307.

    Article  CAS  Google Scholar 

  14. Werner M, Chott A, Fabiano A, Battifora H . Effect of formalin tissue fixation and processing on immunohistochemistry. Am J Surg Pathol 2000; 24: 1016–1019.

    Article  CAS  Google Scholar 

  15. Weber-Matthiesen K, Deerberg J, Poetsch M, Grote W, Schlegelberger B . Numerical chromosome aberrations are present within the CD30+ Hodgkin and Reed–Sternberg cells in 100% of analyzed cases of Hodgkin's disease. Blood 1995; 86: 1464–1468.

    CAS  PubMed  Google Scholar 

  16. Caleo A, Sanchez-Aguilera A, Rodriguez S, Dotor AM, Beltrán L, Larrinoa AF et al. Composite Hodgkin's lymphoma and mantle-cell lymphoma: two clonally unrelated tumors. Am J Surg Pathol 2003, (in press).

  17. Poppema S . The nature of the lymphocytes surrounding Reed–Sternberg cells in nodular lymphocyte predominance and in other types of Hodgkin's disease. Am J Pathol 1989; 135: 351–357.

    CAS  PubMed  PubMed Central  Google Scholar 

Download references


Angel Martinez-Ramírez is a fellow of the Instituto de la Salud Carlos III. Sandra Rodriguez is a fellow of the Fundacion Inocente Inocente. We thank the Immunohistochemistry Unit of the Centro Nacional de Investigaciones Oncologicas (CNIO) for providing monoclonal antibodies and technical support, and the Tumour Bank Network for providing the clinical samples. Finally, we thank Lorraine Tracey for her help.

Author information

Authors and Affiliations


Corresponding author

Correspondence to G Roncador.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Martínez-Ramírez, A., Cigudosa, J., Maestre, L. et al. Simultaneous detection of the immunophenotypic markers and genetic aberrations on routinely processed paraffin sections of lymphoma samples by means of the FICTION technique. Leukemia 18, 348–353 (2004).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:


  • paraffin section
  • lymphomas
  • FISH

This article is cited by


Quick links