Chapter 4
PROTOCOL
FISH on Uncultured Blood or Bone Marrow
THOMAS LIEHR
Introduction
Interphase cytogenetics (= assay to obtain some "cytogenetic information" from interphase nuclei) has been made possible since the advent
of the fluorescence in situ hybridization (FISH) technique (Langer et
al., 1981, Pinkel et al., 1986). It is an approach widely used in diagnostics
since reliable and well estimable probes have been made available for this
technique; i.e. commercially available satellite probes for the centromeric
regions of the human chromosomes to study numerical chromosomal
changes, and cosmids, YACs, BACs or P1-clones suitable for the detection
of translocations, inversions, deletions, duplications or amplifications (for
summary see e.g. Sawyer et al., 1992; Wolfe and Herrington, 1997).
Peripheral blood or bone marrow aspirate are normally subjected to a
short-term culture for 24 to 72 h to obtain metaphase spreads for standard
cytogenetic banding analyses (according to Verma and Babu, 1989). However, in some exceptional cases, direct preparation of uncultured blood or
bone marrow should be given priority (Liehr et al. 1995).
In some cases clinicians send EDTA- or sodium acetate-treated instead
of heparinized blood samples for cytogenetic analysis. It is a well-known
fact that such blood or bone marrow samples cannot be successfully cultured in short-term culture. However, for some FISH analyses, interphase
nuclei are sufficient, as in the detection of microdeletion or microduplication syndromes (i.e. CATCH 22, hereditary neuropathy with liability to
pressure palsies = HNPP, Miller-Diecker syndrome, Cri du Chat syndrome, Williams-Beuren syndrome, Prader-Willi syndrome or CharcotMarie-Tooth disease Type 1A and others), numerical chromosome aberrations detectable by centromeric probes (see Figure 1), or in control of
~ Thomas Liehr, Institut rur Humangenetik, Kollegiengasse 10, Jena, 07743, Germany
(phone +49-3641-935533; fax +49-3641-935502; e-mail i81ith@mti-n.mti.uni-jena.de)
PROTOCOL
FISH on Uncultured Blood or Bone Marrow
THOMAS LIEHR
Introduction
Interphase cytogenetics (= assay to obtain some "cytogenetic information" from interphase nuclei) has been made possible since the advent
of the fluorescence in situ hybridization (FISH) technique (Langer et
al., 1981, Pinkel et al., 1986). It is an approach widely used in diagnostics
since reliable and well estimable probes have been made available for this
technique; i.e. commercially available satellite probes for the centromeric
regions of the human chromosomes to study numerical chromosomal
changes, and cosmids, YACs, BACs or P1-clones suitable for the detection
of translocations, inversions, deletions, duplications or amplifications (for
summary see e.g. Sawyer et al., 1992; Wolfe and Herrington, 1997).
Peripheral blood or bone marrow aspirate are normally subjected to a
short-term culture for 24 to 72 h to obtain metaphase spreads for standard
cytogenetic banding analyses (according to Verma and Babu, 1989). However, in some exceptional cases, direct preparation of uncultured blood or
bone marrow should be given priority (Liehr et al. 1995).
In some cases clinicians send EDTA- or sodium acetate-treated instead
of heparinized blood samples for cytogenetic analysis. It is a well-known
fact that such blood or bone marrow samples cannot be successfully cultured in short-term culture. However, for some FISH analyses, interphase
nuclei are sufficient, as in the detection of microdeletion or microduplication syndromes (i.e. CATCH 22, hereditary neuropathy with liability to
pressure palsies = HNPP, Miller-Diecker syndrome, Cri du Chat syndrome, Williams-Beuren syndrome, Prader-Willi syndrome or CharcotMarie-Tooth disease Type 1A and others), numerical chromosome aberrations detectable by centromeric probes (see Figure 1), or in control of
~ Thomas Liehr, Institut rur Humangenetik, Kollegiengasse 10, Jena, 07743, Germany
(phone +49-3641-935533; fax +49-3641-935502; e-mail i81ith@mti-n.mti.uni-jena.de)