Suspension (S)-FISH, a New Technique for Interphase Nuclei

Steinhaeuser, Ulf; Starke, Heike; Nietzel, Angela; Lindenau, Joerg; Ullmann, P.; Claussen, Uwe; Liehr, Thomas

doi:10.1177/002215540205001216

Cited by 27 publications

(22 citation statements)

References 4 publications

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“…Whole chromosome painting (wcp), partial chromosome painting (pcp), and/or centromeric probes for chromosomes 6, 15, 18, 19, 21, X, and Y [Weise et al 2008] were applied together in suspension-FISH (S-FISH) as previously reported [Iourov et al 2007;Klein et al 2012;Manvelyan et al 2008b;Manvelyan et al 2008c;Manvelyan et al 2009;Steinhaeuser et al 2002]. Images of 3D-preserved interphase nuclei were captured on a Zeiss Axioplan microscope (Car Zeiss, Jena, Germany) and analyzed by the Cell-P (Olympus, Hamburg, Germany) software.…”

Section: Suspension Fish (S-fish)mentioning

confidence: 99%

Small supernumerary marker chromosomes and the nuclear architecture of sperm – a study in a fertile and an infertile brother

Карамышева¹,

Kosyakova

Guediche

et al. 2014

Systems Biology in Reproductive Medicine

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Small supernumerary marker chromosomes (sSMC) are found about four times more frequently in subfertile compared to the general population. The reason for this finding is still unclear. However, a connection of interphase architecture and genome function is suggested. And as we found in a previous study the presence of sSMC influences the nuclear architecture of peripheral blood cells and fibroblasts, we hypothesized that sSMC could have similar effects in sperm cells possibly leading to infertility. Here we applied for the first time 3-dimensional interphase fluorescence in situ hybridization (3D-FISH) to characterize the position of an extrachromosome with respect to its sister-and selected other chromosomes (6, 15, 18, 19, 21, X, and Y) in sperm. Two sSMC carrier brothers with the identical sSMC derived from chromosome 15 were studied. One of the brothers was fertile and the other brother was infertile. Deviations from the normal positioning of chromosomes 21 and Y were seen in both brothers and for chromosomes 19 and X only in the infertile brother. Most striking were high rates of nullisomy and/or disomy for chromosomes 15, including sSMC (15), and 18 exclusively seen in the infertile brother. Overall, further evidence is provided that sSMC influence the nuclear architecture of a cell, including sperm. Further studies are necessary in sperm of fertile and infertile sSMC carriers to elaborate if the detected aneuploidy like that seen in the infertile brother is due to sSMC presence and disturbance of nuclear architecture.

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Section: Suspension Fish (S-fish)mentioning

confidence: 99%

Small supernumerary marker chromosomes and the nuclear architecture of sperm – a study in a fertile and an infertile brother

Карамышева¹,

Kosyakova

Guediche

et al. 2014

Systems Biology in Reproductive Medicine

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“…Fluorescence in situ hybridization (FISH) is recognized as a slide-based imaging application which could benefit greatly by the greater throughput and quantitation of flow cytometry; and several groups have adapted hybridization techniques to cells in suspension. [23][24][25][26] However, the lack of spatial resolution in standard flow cytometry requires the substitution of total probe intensity for spot counting as a means of assessing results, thereby preventing the use of flow cytometry for the analysis of translocations, inversions, or other rearrangements. Though there are certain specific FISH applications that have strong and consistent signals, such as telomeric length analysis or the detection of the presence or absence of a Y chromosome, FISH probe intensity variation can be high and signal intensities often approach the detection limits of standard flow cytometry, reducing the reliability of aneuploidy assessment.…”

Section: High Throughput Extended Depth Of Field Imaging Of Cells Submentioning

confidence: 99%

Cellular Image Analysis and Imaging by Flow Cytometry

Basiji

Ortyn

Liang

et al. 2007

Clinics in Laboratory Medicine

373

288

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SynopsisImaging flow cytometry combines the statistical power and fluorescence sensitivity of standard flow cytometry with the spatial resolution and quantitative morphology of digital microscopy. The technique is a good fit for clinical applications by providing a convenient means for imaging and analyzing cells directly in bodily fluids. Examples are provided of the discrimination of cancerous from normal mammary epithelial cells and the high throughput quantitation of FISH probes in human peripheral blood mononuclear cells. The FISH application will be further enhanced by the integration of extended depth of field imaging technology with the current optical system. Keywordsimaging; flow; cytometry; fluorescence; brightfield; darkfield; multispectral Quantifying Cellular Structure in Health and DiseaseThe eukaryotic cell is a highly structured, three-dimensional object containing a wide range of molecular species. The size, shape, and structure of the cell, as well as the abundance, location, and co-location of any of these constituent biomolecules may be of significance in any given clinical situation or research application. For instance, in hematopoiesis, as cells differentiate and mature, different subsets of molecules are expressed that reflect a specialized functional capacity for that unique cell type (e.g., granulocytes vs. lymphocytes). In general the characterization of this array of constituent molecules by imaging or flow cytometry provides insight into the physiological function of any particular cell or alternatively, pathological changes that may have occurred or accrued. In clinical practice and in research settings, cellular evaluation by imaging technologies and flow cytometry provides significant information reflecting the particular cellular phenotype, both normal and pathological. Microscopy provides a wealth of information, but data acquisition rates are slow and analysis is generally subjective. In flow cytometry, data acquisition is rapid and better suited for the evaluation of pathologies present in low frequency, but the data are only intensity-based, thus lacking the morphology that truly lends credence to the analysis.In addition, the assessment and evaluation of cell samples by imaging and flow cytometric techniques is complicated by a number of factors. For instance, changes in a cell type or aCorresponding author for proofs and reprints:

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“…Multicolor banding (MCB) probe sets for chromosomes 8 and 21 (35,36) were applied in suspension-FISH (S-FISH) as previously reported (17,20,21,37). Images of 3D-preserved interphase nuclei were captured on a Zeiss Axioplan microscope and analyzed by Cell-P (Olympus) software.…”

Section: Multicolor Banding (Mcb) and Suspension Fish (S-fish)mentioning

confidence: 99%

“…Tissue specificity of chromosomal translocations could be due to tissue specific genome organization (11,12) and a positive correlation between spatial proximity of chromosomes/genes in interphase nuclei and translocation frequencies was shown (5,(10)(11)(12)(13). Threedimensional (3D) fluorescence in situ hybridization (FISH) analysis has became a major tool for studying the higher order chromatin organization in the cell nucleus (15)(16)(17)(18)(19)(20)(21).…”

Section: Introductionmentioning

confidence: 99%

Preferred co-localization of chromosome 8 and 21 in myeloid bone marrow cells detected by three dimensional molecular cytogenetics

Manvelyan

Kempf²,

Weise³

et al. 2009

Int J Mol Med

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Abstract. The impact of chromosome architecture in the formation of chromosome aberrations is a recent finding of interphase directed molecular cytogenetic studies. Also positive correlation of translocation frequencies and spatial proximity of chromosomes was described. Thus, disease specific chromosomal translocations could be due to tissue specific genomic organization. However, no three-dimensional interphase fluorescence in situ hybridization (FISH) studies for the nuclear architecture of bone marrow (BM) cells have previously been done. In this study, BM of three secondary acute myelogenous leukemia (AML) cases with trisomy 8 and otherwise normal karyotype were evaluated. Bone marrow cells of one AML and one ALL (acute lymphoblastic leukemia) case, peripheral blood lymphocytes and human sperm, all of them with normal karyotype, served as controls. Multicolor banding (MCB) probes for chromosomes 8 and 21 were applied in suspension-FISH (S-FISH). Interestingly, in myeloid bone marrow cells chromosomes 8 (di-and trisomic) and 21 tended to co-localize with their homologue chromosome(s), rather than to be separated. Thus, the co-localization of chromosomes 8 and 21 might promote a translocation providing a selective advantage of t(8;21) cells in AML-M2. In summary, the concept that tissue specific spatial proximity of chromosomes leads to enhanced translocation frequencies was further supported.

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Suspension (S)-FISH, a New Technique for Interphase Nuclei

Cited by 27 publications

References 4 publications

Small supernumerary marker chromosomes and the nuclear architecture of sperm – a study in a fertile and an infertile brother

Small supernumerary marker chromosomes and the nuclear architecture of sperm – a study in a fertile and an infertile brother

Cellular Image Analysis and Imaging by Flow Cytometry

Preferred co-localization of chromosome 8 and 21 in myeloid bone marrow cells detected by three dimensional molecular cytogenetics

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