Spectral imaging in preconception/preimplantation genetic diagnosis of aneuploidy: multicolor, multichromosome screening of single cells.
OBJECTIVE
Our purpose was to evaluate the utility of spectral imaging for multicolor, multichromosome enumeration in human interphase cell nuclei.


METHODS
Chromosome-specific probes labeled with different fluorochromes or nonfluorescent haptens were obtained commercially or prepared in-house. Metaphase spreads, interphase lymphocytes, or blastomeres cells were hybridized with either 7 or 11 distinctly different probes. Following 46 hr of hybridization, slides were washed and detected using either a filter-based quantitative image processing system (QUIPS) developed in-house or a commercial spectral imaging system.


RESULTS
The filter-based fluorescence microscope system is preferred for simultaneous detection of up to seven chromosome targets because of its high sensitivity and speed. However, this approach may not be applicable to interphase cells when 11 or more targets need to be discriminated. Interferometer-based spectral imaging with a spectral resolution of approximately 10 nm allows labeling of chromosome-specific DNA probes with fluorochromes having greatly overlapping emission spectra. This leads to increases in the number of fluorochromes or fluorochrome combinations available to score unambiguously chromosomes in interphase nuclei.


CONCLUSIONS
Spectral imaging provides a significant improvement over conventional filter-based microscope systems for enumeration of multiple chromosomes in interphase nuclei, although further technical development is necessary in its application to embryonic blastomeres. When applied to preconception/preimplantation genetic diagnosis, presently available probes for spectral imaging are expected to detect abnormalities responsible for 70-80% of spontaneous abortions caused by chromosomal trisomies.
Fung
J
J
Department of Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco 94143-0720, USA.
Hyun
W
W
Dandekar
P
P
Pedersen
R A
RA
Weier
H U
HU
D000782:Aneuploidy; D001757:Blastomeres / Q000737:chemistry*; D015342:DNA Probes; D005260:Female; D005456:Fluorescent Dyes; D005583:Fourier Analysis; D006801:Humans; D007368:Interferometry; D008214:Lymphocytes / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D011247:Pregnancy; D019836:Preimplantation Diagnosis / Q000379:methods*; D013050:Spectrometry, Fluorescence
D015342:DNA Probes; D005456:Fluorescent Dyes