Chapter 23
Detection of Radiosensitivity by Means
of Three-Color FISH
SusANN NEUBAUER
Introduction
Fluorescence in situ hybridization (FISH) using whole chromosome painting (wcp) probes has been shown to be a reliable technique for studying
chromosomal changes. Early investigations in radiation cytogenetics have
been performed using classical cytogenetic analysis of Giemsa-stained
(e.g. Schmid and Bauchinger, 1969) or GTG-banded chromosomes (e.g.
Barrios et al. 1989). The application ofFISH with wcp probes has extended
the spectrum of observed aberrations. Now the reliable detection of translocations, insertions and complex rearrangements is possible in addition
to the well-known acentric fragments, dicentrics and ring chromosomes.
Even though this approach can be done by painting one single chromosome only (e.g. Natarajan et al. 1992) the simultaneous labeling of chromosomes 1,2 and 4 in three different colors allows a more differentiated
analysis of complex chromosome aberrations. Using this three-color FISH
technique where the three selected chromosomes represent about 22% of
the DNA-content of the human genome a sensitive method for studying
radiation-induced chromosomal aberrations is available (Gebhart et al.
1996).
Up to now, radiation induced chromosomal aberrations were analyzed
using different specific wcp probes (Natarajan et al. 1992), by single-colorFISH studying each human chromosome separately (Cigarran et al. 1998),
and even using multi-color FISH (Greulich et al. 2000). The principal idea
of the three-color FISH method presented here was to develop a simple
and rapid assay for the detection of increased radiosensitivity in cancer
patients (Neubauer et al. 1996). Performing the three-color FISH method,
it is possible to detect and quantify increased radiosensitivity on the one
Susann Neubauer, Gynakologen-Medizinische Genetik, Bankgasse 3, Niirnberg, 90402,
Germany (phone 0049-911-2061024; fax 0049-911-2061031;
e-mail genetik@praenatal.com)
PROTOCOL
Detection of Radiosensitivity by Means
of Three-Color FISH
SusANN NEUBAUER
Introduction
Fluorescence in situ hybridization (FISH) using whole chromosome painting (wcp) probes has been shown to be a reliable technique for studying
chromosomal changes. Early investigations in radiation cytogenetics have
been performed using classical cytogenetic analysis of Giemsa-stained
(e.g. Schmid and Bauchinger, 1969) or GTG-banded chromosomes (e.g.
Barrios et al. 1989). The application ofFISH with wcp probes has extended
the spectrum of observed aberrations. Now the reliable detection of translocations, insertions and complex rearrangements is possible in addition
to the well-known acentric fragments, dicentrics and ring chromosomes.
Even though this approach can be done by painting one single chromosome only (e.g. Natarajan et al. 1992) the simultaneous labeling of chromosomes 1,2 and 4 in three different colors allows a more differentiated
analysis of complex chromosome aberrations. Using this three-color FISH
technique where the three selected chromosomes represent about 22% of
the DNA-content of the human genome a sensitive method for studying
radiation-induced chromosomal aberrations is available (Gebhart et al.
1996).
Up to now, radiation induced chromosomal aberrations were analyzed
using different specific wcp probes (Natarajan et al. 1992), by single-colorFISH studying each human chromosome separately (Cigarran et al. 1998),
and even using multi-color FISH (Greulich et al. 2000). The principal idea
of the three-color FISH method presented here was to develop a simple
and rapid assay for the detection of increased radiosensitivity in cancer
patients (Neubauer et al. 1996). Performing the three-color FISH method,
it is possible to detect and quantify increased radiosensitivity on the one
Susann Neubauer, Gynakologen-Medizinische Genetik, Bankgasse 3, Niirnberg, 90402,
Germany (phone 0049-911-2061024; fax 0049-911-2061031;
e-mail genetik@praenatal.com)
PROTOCOL