34 Simultaneous Fluorescence Immunophenotyping and FISH (FICTION)
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or indirect fluorescence in situ hybridization (FISH) is performed using
appropriately modified DNA probes. Using appropriate fLlter sets, the immunophenotype and the hybridization signals can be evaluated simultaneously under a fluorescence microscope (for review see Siebert and
Weber-Matthiesen 1997; Siebert and ScWegelberger 1997).
For example, the following objectives can be studied using the FICTION technique:
- The immunophenotypic characterization of tumor cell clones with defined chromosome aberrations (Haferlach et al. 1996, 1997; Zhang et al.
1999, 2000);
- The definition of the differentiation state present in the tumor cell
clone (Weber-Matthiesen et al. 1995a);
- The identification of the most immature precursor cells within the tumor clone representing the cell population from which the tumor clone
may have originated (Zhang et al. 1996);
- The immunophenotypic characterization of bystander cells with a normal chromosome constitution (Weber-Matthiesen et al. 1995b);
- The chromosomal characterization of small tumor cell populations
with a selective immunophenotype within a background of normal
cells (Weber-Matthiesen et al. 1995c, 1996).
Materials
- AMCA-conjugated avidin (Jackson 003-150-083)
- Biotin-labeled goat anti-avidin antibody (Vector BA-0300)
- Class-matched mouse antibodies IgGl, IgG2a, IgG2b, IgM (Immunotech 0571, 0572, 1266, 1268)
- Human Cot-l DNA (Gibco BRLlLife Technologies 15279-011)
- Cy3-conjugated avidin (Jackson 003-160-083)
- Cy3-conjugated goat anti-mouse antibody (Jackson 115-165-068)
- Cy3-conjugated rabbit anti-goat antibody (Jackson 305-165-045)
- Cy3-conjugated donkey anti-rabbit antibody (Jackson 711-165-152)
- Cy3-conjugated rabbit anti-mouse antibody (Jackson 315-165-003)
Reagents