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.
Martínez-Ramírez
A
A
Department of Human Genetics, Centro Nacional Investigaciones Oncologicas (CNIO), Madrid, Spain.
Cigudosa
J C
JC
Maestre
L
L
Rodríguez-Perales
S
S
Haralambieva
E
E
Benítez
J
J
Roncador
G
G
D002021:Buffers; D020732:Cytogenetic Analysis / Q000379:methods* / Q000592:standards; D019085:Fluorescent Antibody Technique, Direct; D006801:Humans; D016130:Immunophenotyping / Q000379:methods* / Q000592:standards; D017404:In Situ Hybridization, Fluorescence; D008223:Lymphoma / Q000473:pathology*; D015335:Molecular Probes; D016612:Paraffin Embedding; D012680:Sensitivity and Specificity
D002021:Buffers; D015335:Molecular Probes