细胞功能测定

It is well known that chromosome in situ hybridization allows the unequivocal identification of targeted human somatic chromosomes. Different fluorescence in situ hybridization (FISH) techniques have been developed over the years and, following mitotic studies, meiotic anal ...

Interphase cytogenetics using archival tissue samples is a straightforward approach that can be used to obtain cytogenetic information from nuclei of solid tissue samples. It is the only tool available for investigating specific numerical chromosomal aberrations, chromosom ...

Mosaic karyotypes are present in at least 0.3–1% of the clinical cases analyzed by banding cytogenetics. It is well known that the pattern of mosaicism can vary greatly in different tissue types from the same patient. However, normally a maximum of two different tissues from a child or an adult are studi ...

Archival cytogenetic slides of unique clinical and tumorgenetic cases embedded in Eukitt� or Canada balsam can be made available for molecular cytogenetic studies using the recovery technique presented here. Metaphase and interphase FISH can be used to reevaluate previously dete ...

Tumor cytogenetics is an essential tool for diagnosis and prognosis differentiation, especially in malignancies of the peripheral blood. As metaphase spreads are sometimes difficult to obtain in leukemia and lymphoma preparations, interphase cytogenetics is currently the m ...

Fluorescence in situ hybridization (FISH) in prenatal diagnostics permits the demonstration of certain numeric chromosomal aneuploidies in amniotic cells within 24h, in contrast to conventional methods, which take 1–3 weeks. The experience of using this FISH method has been compa ...

Peripheral blood, amniocytic fluid, chorion, and fibroblasts are the tissues most frequently used for chromosome studies. All four of them are relatively easy to obtain and simply brought into short-term culture, and metaphase spreads can be prepared within a relatively short time. Such m ...

Complex FISH probes comprising large spans of genomic DNA always contain a high amount of dispersed repetitive sequences that hamper the visualization of the specific signal of interest. To overcome this problem, different approaches have been elaborated that depend on the type of exper ...

Fluorescence in situ hybridization (FISH) is strongly dependent on good hardware, i.e., the microscope and the filter sets used. Here an introduction to fluorescence, fluorescence spectra and fluorescent dyes, as well as filters and their characteristics, is given. Hints on how to choose the ...

Microscopy is indisputably an essential technique for all laboratories that use fluorescence in situ hybridization (FISH). Although visual analysis of FISH results is still used, a growing number of molecular cytogenetic approaches require the application of specific imaging s ...

The use of highly specific synthetic oligonucleotide (ODN) probes, which have superior hybridization kinetics and lower manufacturing costs compared to traditional genomic probes, is one approach that can be used to perform chromosome enumeration on interphase nuclei by FISH. ODNs ...

Peptide nucleic acid in situ labeling (PNA-FISH) techniques provide an attractive alternative to conventional FISH and PRINS procedures for chromosomal in situ detection. PNA probes are synthetic DNA analogs with uncharged polyamide backbones. The PNA labeling reaction presen ...

Besides the well-known applications of bacterial artificial chromosomes (BACs) in classical molecular genetics, BACs are also used for molecular cytogenetic studies. BACs, as well as other locus-specific probes like cDNA, plasmids, cosmids, fosmids, P1-clones or yeast artifici ...

FISH with whole-chromosome or region-specific painting probes made from either flow-sorted or microdissected chromosomes has revolutionized cytogenetics. The generation of paints from flow-sorted chromosomes relies on the use of an expensive and sophisticated fluoresc ...

In this chapter, the two basic principal techniques of molecular cytogenetics are outlined, i.e., fluorescence in situ hybridization (FISH) and primed in situ hybridization (PRINS). Basic protocols are provided, which can be varied and adapted according to specific application(s).

Molecular cytogenetics is a new branch of chromosome research that is just is just over 25 years old. Current applications of fluorescence in situ hybridization (FISH) in diagnostics and research are summarized in this chapter. An overview of both DNA- and RNA-based in situ hybridization is pr ...

Comparative genomic hybridization (CGH) is a molecular cytogenetic technique developed for the analysis of chromosome imbalance in tumors and constitutional chromosome abnormalities. It is based on the analysis of genomic DNA and has the advantage over conventional karyotypi ...

Preimplantation genetic diagnosis for single gene disorders is usually performed using polymerase chain reaction (PCR)-based methodologies modified for use in single cells. At present, single cell PCR tests require costly and time-consuming development and validation of hig ...

Transient overexpression of wild-type or mutant cell cycle regulatory factors is a quick and relatively easy means to derive important information regarding the function of the respective factor in regulating cell cycle checkpoint pathways. The general approach for these studies is ...

Injuries to the peripheral nervous system affect 1 in 1,000 individuals each year. The implication of sustaining such an injury is considerable with loss of sensory and/or motor function. The economic implications too are extensive running into millions of pounds (or dollars) annually for ...