29 Spectral Imaging Analysis of Metaphase and Interphase Cells
383
Comments
Purchasing the whole chromosome painting probes for spectral karyotyping or m-fish from one of the commercial suppliers might be more efficient than attempting to synthesize painting probes inhouse. If metaphase
or other target cells are dense enough, a small coverslip can be used to
conserve valuable probe material. For a 12 mm circular cover glass, for
example, 2 III of probe is often sufficient.
Applications
SKY on metaphase spreads
SKY allows the chromosomes to be "painted» in different colors and, thus,
is an excellent tool to analyze complex rearrangements as well as to detect
hidden chromosomal abnormalities that might be present in tumor tissues. Often chromosome analysis in tumors is complicated by polyploid
genomes. SKY permits a detailed analysis of marker chromosomes in the
complex rearrangements and tri- or tetraploidy cases (Fig. 1). SKY has
been applied in several cancer studies (Schrock 1996, Zitzelsberger
1999), prenatal diagnosis (Ning 1999), and for analysis of human oocytes,
polar bodies biopsied from human oocytes (Marquez 1998), artificially
condensed chromatin of blastomeres (Willadsen 1999), and mouse karyotyping (Liyanage 1996).
Sim on interphase cells
Interphase FISH has widely been used to analyze chromosome or gene
copy numbers in tumor tissues, interphase lymphocytes or for the rapid
prenatal genetic diagnosis of aneuploidy. Commercially available kits
presently allow enumeration of just five chromosomes in interphase cells
using filter-based fluorescence microscope systems [X, Y, 13, 18, 21
(AneuVysion assay kit, Vysis) or 13, 16, 18,21,22 (MultiVysion PB multicolor probe panel, Vysis)). Repeated hybridizations on single interphase
cells can be applied to score additional chromosomes. Signal overlap,
however, may cause severe problems in domain scoring and diagnosis
of interphase cells. Therefore, Slm techniques were developed to recognize
the majority of numerical chromosome aberrations in the few available
fetal or embryonic interphase cells. Using Slm, it is now possible to simul-
383
Comments
Purchasing the whole chromosome painting probes for spectral karyotyping or m-fish from one of the commercial suppliers might be more efficient than attempting to synthesize painting probes inhouse. If metaphase
or other target cells are dense enough, a small coverslip can be used to
conserve valuable probe material. For a 12 mm circular cover glass, for
example, 2 III of probe is often sufficient.
Applications
SKY on metaphase spreads
SKY allows the chromosomes to be "painted» in different colors and, thus,
is an excellent tool to analyze complex rearrangements as well as to detect
hidden chromosomal abnormalities that might be present in tumor tissues. Often chromosome analysis in tumors is complicated by polyploid
genomes. SKY permits a detailed analysis of marker chromosomes in the
complex rearrangements and tri- or tetraploidy cases (Fig. 1). SKY has
been applied in several cancer studies (Schrock 1996, Zitzelsberger
1999), prenatal diagnosis (Ning 1999), and for analysis of human oocytes,
polar bodies biopsied from human oocytes (Marquez 1998), artificially
condensed chromatin of blastomeres (Willadsen 1999), and mouse karyotyping (Liyanage 1996).
Sim on interphase cells
Interphase FISH has widely been used to analyze chromosome or gene
copy numbers in tumor tissues, interphase lymphocytes or for the rapid
prenatal genetic diagnosis of aneuploidy. Commercially available kits
presently allow enumeration of just five chromosomes in interphase cells
using filter-based fluorescence microscope systems [X, Y, 13, 18, 21
(AneuVysion assay kit, Vysis) or 13, 16, 18,21,22 (MultiVysion PB multicolor probe panel, Vysis)). Repeated hybridizations on single interphase
cells can be applied to score additional chromosomes. Signal overlap,
however, may cause severe problems in domain scoring and diagnosis
of interphase cells. Therefore, Slm techniques were developed to recognize
the majority of numerical chromosome aberrations in the few available
fetal or embryonic interphase cells. Using Slm, it is now possible to simul-