Spectral karyotyping

Garini, Yuval; Macville, Merryn; Manoir, Stanislas du; Buckwald, Robert A; Lavi, Moshe; Katzir, Nir; Wine, David; Bar‐Am, Irit; Schröck, Evelin; Cabib, Dario; Ried, Thomas

doi:10.1002/1361-6374(199606)4:2<65::aid-bio4>3.3.co;2-4

Cited by 97 publications

(61 citation statements)

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“…This approach has for some years found success combined with imaging spectroscopy to perform, e.g. spectral karyotyping (Garini et al 1996). Briefly, the method uses the presence or absence of n dyes to label 2 n K1 objects.…”

Section: Resultsmentioning

confidence: 99%

“…Systems for generating spectral images with two spatial dimensions (x, y, l and I ) have been described. These include imaging spectrographs combined with stage scanning, Hadamard ( Hanley et al 1999) and Fourier (Garini et al 1996) multiplexed systems, tunable filters (Levenson 2006) and confocal laser scanning microscopes with microspectroscopic hardware (Rück et al 2007). The latter three approaches are available commercially.…”

Section: Spectral Imagesmentioning

confidence: 99%

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Spectrally resolved fluorescent lifetime imaging

Hanley

2008

J. R. Soc. Interface.

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Placing an imaging spectrograph or related components capable of generating a spectrum between a microscope and the image intensifier of a conventional fluorescence lifetime imaging (FLIM) system creates a spectrally resolved FLIM (SFLIM). This arrangement provides a number of opportunities not readily available to conventional systems using bandpass filters. The examples include: simultaneous viewing of multiple fluorophores; tracking of both the donor and acceptor; and observation of a range of spectroscopic changes invisible to the conventional FLIM systems. In the frequency-domain implementation of the method, variation in the fractional contributions from different fluorophores along the wavelength dimension can behave as a surrogate for a frequency sweep or spatial variations while analysing fluorophore mixtures. This paper reviews the development of the SFLIM method, provides a theoretical and practical overview of frequency-domain SFLIM including: presentation of the data; manifestations of energy transfer; observation of multiple fluorophores; and the limits of single frequency methods.

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Section: Resultsmentioning

confidence: 99%

Section: Spectral Imagesmentioning

confidence: 99%

Spectrally resolved fluorescent lifetime imaging

Hanley

2008

J. R. Soc. Interface.

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“…After in situ hybridization images were acquired using an epi¯uorescence microscope (DMRXA, Leica, Wetzlar, Germany) connected to an imaging interferometer (SD200, Applied Spectral Imaging, Migdal HaEmek, Israel). Chromosomes were unambiguously identi®ed using a spectral classi®cation algorithm that results in the assignment of a separate classi®cation color to all pixels with identical spectra (Garini et al, 1996). Chromosome aberrations were de®ned using the nomenclature rules from the International Committee on Standardized Genetic Nomenclature for Mice (Davisson, 1994).…”

Section: Molecular Cytogenetic Analysesmentioning

confidence: 99%

Centrosome abnormalities, recurring deletions of chromosome 4, and genomic amplification of HER2/neu define mouse mammary gland adenocarcinomas induced by mutant HER2/neu

et al. 2002

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The conditional expression of activated HER2/neu gene under its endogenous promoter in the mammary epithelium of the mouse results in accelerated lobular development and focal mammary tumors. Carcinogenesis, however, requires ampli®cation and considerably increased expression levels of oncogenic neu. Deducing from the multiple genetic aberrations required for human breast cancer to develop, we hypothesized that in addition to the over-expression of an activated HER2/ neu, secondary aberrations would occur. We have therefore conducted a genomic screen for chromosomal imbalances and translocations using comparative genomic hybridization and spectral karyotyping. The results reveal a moderate degree of chromosomal instability and micronuclei formation in short-term cultures established from primary tumors. Genomic instability appears to be linked to the ampli®cation of functional centrosomes, a phenomenon that we frequently observed in other tumor types. Seventy per cent of the tumors revealed genomic ampli®cation of HER2/neu, often in the form of double minute chromosomes, which correlated with recurring loss of mouse chromosome 4D-E, a region that is orthologous to distal human chromosome 1p. It is likely that this region contains putative tumor suppressor genes whose inactivation is required for tumor formation in this model of human breast cancer.

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“…Metaphase spreads were stained with 4Ј,6-diamidino-2-phenylindole (DAPI) solution (150 ng/ml in 2 × SSC) and covered with antifade solution (Vectashield mounting medium; Vector Laboratories, Burlingame, Calif.). Metaphase images were acquired by use of a SpectraCube system (ASI) and analyzed with the SKYView imaging software [17]. Aberrations were described according to the ISCN nomenclature [15].…”

Section: Spectral Karyotypingmentioning

confidence: 99%