Chapter 34
PROTOCOL
Simultaneous Fluorescence Immunophenotyping
and FISH (FICTION)
BRIGITTE SCHLEGELBERGER, STEFAN GESK, YANMING ZHANG,
and REINER SIEBERT
Introduction
In this chapter, the FICTION (Fluorescence Immunophenotyping and Interphase Cytogenetics as a Tool for Investigation of Neoplasms) technique
will be detailed. This method has been described in 1992 by Weber-Matthiesen et al. as the first technique to combine immunophenotyping and
FISH. In contrast to previous methods like the morphology-antibodychromosome (MAC) technique (Teerenhovi et al. 1984; Schlegelberger
et al. 1992), the FICTION technique can be applied on interphase cells,
which is of great advantage especially for tumor cytogenetics. To briefly
introduce the technique: In general, conventional unstained peripheral
blood or bone marrow smears, imprints and cytospin slides from fresh
tissue and cryostat sections are applicable for FICTION. In order to preserve cellular antigens, preparing high quality slides is one of the critical
steps for successfully performing FICTION. Freshly prepared slides should
be air dried overnight at room temperature. Thereafter, slides can be processed immediately or stored airtight at -80
a C for years. For the FICTION
procedure, slides are fixed in acetone and then immunophenotyped by a
direct or indirect fluorescence detection system. Immunophenotyping is
followed by a fixation step to preserve the fluorescent immunostaining
during the harsh hybridization procedure. After fixation, standard direct
~ Brigitte Schlegelberger, Medizinische Hochschule Hannover, Institut filr Zell- und
Molekularpathologie, Carl-Neuberg-Str. 1, Hannover, 30625, Germany (phone +49511-532-4522; fax +49-511-532-4521; e-mail Schlegelberger.Brigitte@mh-hannover.de)
Stefan Gesk, Universitat Kiel, Institut filr Humangenetik, Schwanenweg 24, Kiel, 24105,
Germany
Yanming Zhang, Universitat Kiel, Institut filr Humangenetik, Schwanenweg 24, Kiel,
24105, Germany, Present address: University of Chicago, Medical Center, Section of
Hematology/Oncology, 5841 S. Maryland Ave. MC21I5, Chicago, 11,60637-1470, USA
Reiner Siebert, Universitat Kiel, Institut filr Humangenetik, Schwanenweg 24, Kiel,
24105, Germany
PROTOCOL
Simultaneous Fluorescence Immunophenotyping
and FISH (FICTION)
BRIGITTE SCHLEGELBERGER, STEFAN GESK, YANMING ZHANG,
and REINER SIEBERT
Introduction
In this chapter, the FICTION (Fluorescence Immunophenotyping and Interphase Cytogenetics as a Tool for Investigation of Neoplasms) technique
will be detailed. This method has been described in 1992 by Weber-Matthiesen et al. as the first technique to combine immunophenotyping and
FISH. In contrast to previous methods like the morphology-antibodychromosome (MAC) technique (Teerenhovi et al. 1984; Schlegelberger
et al. 1992), the FICTION technique can be applied on interphase cells,
which is of great advantage especially for tumor cytogenetics. To briefly
introduce the technique: In general, conventional unstained peripheral
blood or bone marrow smears, imprints and cytospin slides from fresh
tissue and cryostat sections are applicable for FICTION. In order to preserve cellular antigens, preparing high quality slides is one of the critical
steps for successfully performing FICTION. Freshly prepared slides should
be air dried overnight at room temperature. Thereafter, slides can be processed immediately or stored airtight at -80
a C for years. For the FICTION
procedure, slides are fixed in acetone and then immunophenotyped by a
direct or indirect fluorescence detection system. Immunophenotyping is
followed by a fixation step to preserve the fluorescent immunostaining
during the harsh hybridization procedure. After fixation, standard direct
~ Brigitte Schlegelberger, Medizinische Hochschule Hannover, Institut filr Zell- und
Molekularpathologie, Carl-Neuberg-Str. 1, Hannover, 30625, Germany (phone +49511-532-4522; fax +49-511-532-4521; e-mail Schlegelberger.Brigitte@mh-hannover.de)
Stefan Gesk, Universitat Kiel, Institut filr Humangenetik, Schwanenweg 24, Kiel, 24105,
Germany
Yanming Zhang, Universitat Kiel, Institut filr Humangenetik, Schwanenweg 24, Kiel,
24105, Germany, Present address: University of Chicago, Medical Center, Section of
Hematology/Oncology, 5841 S. Maryland Ave. MC21I5, Chicago, 11,60637-1470, USA
Reiner Siebert, Universitat Kiel, Institut filr Humangenetik, Schwanenweg 24, Kiel,
24105, Germany