Chapter 29
Spectral Imaging Analysis of Metaphase
and Interphase Cells
JINGLY FUNG, HEINZ-ULLI G. WEIER,
ROGER A. PEDERSEN, and HORST F. ZITZELSBERGER
Introduction
Spectral imaging is a novel technology combining spectroscopy and imaging for the analysis of fluorescence in situ hybridization (FISH). This system combines the techniques of fluorescence microscopy, charge-coupled
device (CCD) camera, Fourier spectroscopy, and powerful computer software. It had initially been developed to screen human metaphase chromosome spreads after simultaneously hybridizing the chromosomes with 24
chromosome-specific whole chromosome painting probes. This particular
application, termed Spectral Karyotyping (SKY) (Garini 1996), is mostly
used in karyotype analysis of constitutional chromosome abnormalities,
particularly in tumor studies.
In some circumstances, the target cells are few and metaphase spreads
are very difficult to obtain. Blastomeres from human embryos for preimplantation genetic diagnosis (PGD), uncultured amniocytes from amniocentesis, and fetal cells from maternal blood for prenatal diagnosis are
examples. Because aneuploidy of any of the 24 chromosomes may have
~ Jingly Fung, University of California, Department of Obstetrics, Gynecology and
Reproductive Sciences, Reproductive Genetics Unit, Box 0720, 533 Parnassus, San
Francisco, CA, 94143-0720, USA (phone +415-476-8517; fax +415-476-6145;
e-mail jlfung@itsa.ucsf.edu)
University of California, Department of Subcellular Structures, Life Sciences Division,
E. O. Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA
Heinz-Um G. Weier, University of California, Department of Subcellular Structures,
Life Sciences Division, E. O. Lawrence Berkeley National Laboratory, Berkeley, CA,
94720, USA
Roger A. Pedersen, University of California, Department of Obstetrics, Gynecology
and Reproductive Sciences, Reproductive Genetics Unit, Box 0720, 533 Parnassus, San
Francisco, CA, 94143-0720, USA
Horst F. Zitzelsberger, gsf Forschungszentrum fiir Umwelt- und Gesundheit,
Arbeitsgruppe Zytogenetik, Oberschleissheim, 85758, Germany
PROTOCOL
Spectral Imaging Analysis of Metaphase
and Interphase Cells
JINGLY FUNG, HEINZ-ULLI G. WEIER,
ROGER A. PEDERSEN, and HORST F. ZITZELSBERGER
Introduction
Spectral imaging is a novel technology combining spectroscopy and imaging for the analysis of fluorescence in situ hybridization (FISH). This system combines the techniques of fluorescence microscopy, charge-coupled
device (CCD) camera, Fourier spectroscopy, and powerful computer software. It had initially been developed to screen human metaphase chromosome spreads after simultaneously hybridizing the chromosomes with 24
chromosome-specific whole chromosome painting probes. This particular
application, termed Spectral Karyotyping (SKY) (Garini 1996), is mostly
used in karyotype analysis of constitutional chromosome abnormalities,
particularly in tumor studies.
In some circumstances, the target cells are few and metaphase spreads
are very difficult to obtain. Blastomeres from human embryos for preimplantation genetic diagnosis (PGD), uncultured amniocytes from amniocentesis, and fetal cells from maternal blood for prenatal diagnosis are
examples. Because aneuploidy of any of the 24 chromosomes may have
~ Jingly Fung, University of California, Department of Obstetrics, Gynecology and
Reproductive Sciences, Reproductive Genetics Unit, Box 0720, 533 Parnassus, San
Francisco, CA, 94143-0720, USA (phone +415-476-8517; fax +415-476-6145;
e-mail jlfung@itsa.ucsf.edu)
University of California, Department of Subcellular Structures, Life Sciences Division,
E. O. Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA
Heinz-Um G. Weier, University of California, Department of Subcellular Structures,
Life Sciences Division, E. O. Lawrence Berkeley National Laboratory, Berkeley, CA,
94720, USA
Roger A. Pedersen, University of California, Department of Obstetrics, Gynecology
and Reproductive Sciences, Reproductive Genetics Unit, Box 0720, 533 Parnassus, San
Francisco, CA, 94143-0720, USA
Horst F. Zitzelsberger, gsf Forschungszentrum fiir Umwelt- und Gesundheit,
Arbeitsgruppe Zytogenetik, Oberschleissheim, 85758, Germany
PROTOCOL