A5006408384- Advanced biosensing and bioanalysis techniques
- Genomic variations and chromosomal abnormalities
- Molecular Biology Techniques and Applications
- Chromosomal and Genetic Variations
- DNA and Nucleic Acid Chemistry
- RNA and protein synthesis mechanisms
- Cytomegalovirus and herpesvirus research
- Genomics and Chromatin Dynamics
- Gene expression and cancer classification
- Chronic Lymphocytic Leukemia Research
- RNA modifications and cancer
- Herpesvirus Infections and Treatments
- Mitochondrial Function and Pathology
- Lymphoma Diagnosis and Treatment
- RNA Research and Splicing
- RNA Interference and Gene Delivery
- Virus-based gene therapy research
- Bacteriophages and microbial interactions
- Acute Myeloid Leukemia Research
- Cell Image Analysis Techniques
- Single-cell and spatial transcriptomics
- CRISPR and Genetic Engineering
- Biotin and Related Studies
- Cancer Genomics and Diagnostics
- DNA Repair Mechanisms
Leiden University
1993-2015
Leiden University Medical Center
2000-2012
National Human Genome Research Institute
2000
Cancer Genetics (United States)
2000
National Institutes of Health
2000
Royal Marsden Hospital
1998
The Netherlands Cancer Institute
1998
Universidad de Salamanca
1995-1997
Complejo Hospitalario de Salamanca
1997
University of British Columbia
1997
Vertebrate chromosomes terminate in variable numbers of T 2 AG 3 nucleotide repeats. In order to study telomere repeats at individual chromosomes, we developed novel, quantitative fluorescence situ hybridization procedures using labeled (C TA ) peptide nucleic acid and digital imaging microscopy. Telomere intensity values from metaphase cultured human hematopoietic cells decreased with the replication history cells, varied up six-fold within a metaphase, were similar between sister chromatid...
The ultra-long telomeres that have been observed in mice are not accordance with the concept critical telomere shortening is related to aging and immortalization. Here, we used quantitative fluorescence situ hybridization estimate (T 2 AG 3 ) n lengths of individual various mouse strains. Telomere were very heterogeneous, but specific chromosomes bone marrow cells skin fibroblasts from had similar lengths. We shortest around 10 kb length, indicating each cell has a few within range human...
The translocation (6;9)(p23;q34) in acute nonlymphocytic leukemia results the formation of a highly consistent dek-can fusion gene. Translocation breakpoints invariably occur single introns dek and can, which were named icb-6 icb-9, respectively. In case undifferentiated leukemia, breakpoint was detected icb-9 whereas no could be dek. Genomic cDNA cloning showed that instead dek, different gene fused to set. set encodes transcripts 2.0 2.7 kb result from use alternative polyadenylation...
A method for multiple fluorescence in situ hybridization is described allowing the simultaneous detection of more than three target sequences with only fluorescent dyes (FITC, TRITC, AMCA), respectively emitting green, red, and blue. This procedure based on labeling (DNA) probes one hapten visualisation colors. The possibility to detect targets simultaneously important prenatal diagnosis numerical and/or structural chromosome aberrations tumor diagnosis. It may form basis an banding technique.
Journal Article Ultra-sensitive FISH using peroxidase-mediated deposition of biotin- or fluorochrome tyramides Get access A.K. Raap, Raap * Department Cytochemistry and Cytometry, Medical Faculty, Leiden UniversityWassenaarseweg 72, 2333 AL Leiden, The Netherlands *To whom correspondence should be addressed Search for other works by this author on: Oxford Academic PubMed Google Scholar M.P.C. van de Corput, Corput R.A.M. Vervenne, Vervenne R.P.M. Gijlswijk, Gijlswijk H.J. Tanke, Tanke J....
Peripheral blood leukocyte samples (n = 458) of 24 bone marrow transplant and 52 kidney patients were examined weekly for the presence human cytomegalovirus (HCMV) using an improved culture technique (DEAFF; detection early antigen fluorescent foci). In total 5 (21%) 11 developed a viremia. Patients' investigated HCMV DNA in vitro amplification technique, polymerase chain reaction (PCR). From statistically evaluable viremic 13), 110 analyzed. these patients, DEAFF PCR led to identical...
Abstract A method is described for visualizing three nucleic acid sequences simultaneously by in situ hybridization using a new blue immunofluorescent label, amino methyl coumarin acetic (AMCA), combination with green and red fluorescing FITC TRITC. Three chromosome‐specific repetitive probes labeled either acetyl fluorene (AAF), mercury, or biotin were hybridized to metaphase chromosomes prepared from human blood lymphocytes interphase tumor nuclei. Conditions the combined use of...
The peroxidase-mediated deposition of hapten- and fluorochrome-labeled tyramides has recently been shown to increase the sensitivity immunofluorescence fluorescence in situ hybridization techniques. We have evaluated a number red, green, blue fluorescent for detection antigens tissue sections cytospin preparations horseradish peroxidase-labeled probes hybridized cells chromosomes. With few exceptions, all tyramide-based methods provided considerable compared conventional FISH methods.
We have used fluorescein-11-dUTP in a nick-translation format to produce fluoresceinated human nucleic acid probes. After In situ hybridization of DNAs metaphase chromosomes, the detection sensitivity was found be 50 -100 kb. The feasibility and increase microscopic imaging hybridized, DNA with an integrating solid state camera for rapid cosmld mapping is illustrated. Combination biotinated dlgoxigenlnated allowed easy performance triple fluorescence hybridization. potential these techniques...
Double fluorescence in situ hybridization was used to detect Philadelphia (Ph) chromosomes interphase nuclei and metaphases of patients with chronic myeloid leukemia. Application cosmid probes for 3’ ABL 5’ BCR sequences gave better results than libraries 9 22. The present approach may provide an alternative method monitoring minimal residual disease Ph<sup>+</sup> CML patients.
To improve DNA resolution of fluorescence in situ hybridization we have adapted a nuclear extraction technique, resulting highly extended loops arranged around the matrix halo-like structure. In signals from alphoid and cosmid DNAs appear as beads-on-a-string, which, according to preliminary experiments, results association individual probe fragments. By multicolor hybridizations been able determine relative map position easily detect 10 kb overlap between clones, each which shows linear...
The human genome is made up of long DNA segments, the isochores, which are compositionally homogeneous and can be subdivided into a small number families characterized by different G+C levels. Chromosome in situ suppression hybridization (in excess unlabeled added to suppress repeated sequences present probe, enabling enhanced observation single-copy sequences) fractions an increasing level was carried out determine distribution "single-copy" corresponding isochore L1+L2, H1, H2, H3 on...
We have applied two-colour fluorescence in situ hybridization (FISH) to DNA fibers and combined it with digital imaging microscopy for the mapping of large cosmid contigs. The technique was validated using a set unique plasmids contig both originating from thyroglobulin (Tg) gene previously mapped by restriction analysis. resolution proved be close theoretical lower limit ˜1 kb, ranging ≥400 kb. Subsequently 400 kb derived DMD-YAC directly Fiber-FISH. resulting map is full agreement map....
To evaluate the clinical relevance of genomic aberrations in primary cutaneous large B-cell lymphoma (PCLBCL).Skin biopsy samples 31 patients with a PCLBCL classified as either follicle center (PCFCL; n = 19) or PCLBCL, leg type (n 12), according to WHO-European Organisation for Research and Treatment Cancer (EORTC) classification, were investigated using array-based comparative hybridization, fluorescence situ hybridization (FISH), examination promoter hypermethylation.The most recurrent...
The cellular origin of the sarcomatous component gliosarcomas is controversial. It not clear if sarcoma arises in transition from glial cells that comprise gliomatous or independently non-neoplastic mesenchymal tumor stroma. Using comparative genomic hybridization (CGH) along with cytogenetic analysis, fluorescence situ (FISH) and polymerase chain reaction (PCR) analysis microsatellite allelic imbalance, we have evaluated genetic alterations components five gliosarcomas. element was grade 4...
Fluorescence in situ hybridization (FISH) is now widely used for the localization of genomic DNA fragments, and identification chromosomes by painting. We show that half chromosomal complement can be painted twelve different colors using human chromosome specific libraries carrying three distinct labels mixed multiple ratios. The photographs are 'real' color rather than 'colorized'. painting technique described here small or complex rearrangements marker humans any other species which well...
The hSNF5 subunit of human SWI/SNF ATP-dependent chromatin remodeling complexes is a tumor suppressor that inactivated in malignant rhabdoid tumors (MRTs). Here, we report loss function MRT-derived cells leads to polyploidization and chromosomal instability. Re-expression restored the coupling between cell cycle progression ploidy checkpoints. In contrast, cancer-associated mutants harboring specific single amino acid substitutions exacerbated poly- aneuploidization, due abrogated chromosome...