In the 1930s, the discovery of a simple method for the isolation and detailed microscopic observation of the banded structures that lie within the nuclei of salivary gland cells of Drosophila melanogaster was soon followed by the realization that these structures were in fact a highly amplif ...

Chromosomal in situ suppression (CISS) hybridization with flow-sorted chromosome libraries (FSCL) has provided a very powerful tool in human cytogenetics. This technique allows the painting of specific chromosomes in metaphase spreads and in interphase nuclei (1–4). The useful ...

Detecting viral infections depends largely on serological methods to demonstrate the presence of viral circulating antibodies and molecular techniques to identify viral genomes in tissue extracts. Although sensitive, none of these provides the simultaneous morphological ...

Unlike other applications of in situ hybridization (ISH),where the target is likely to be chromosomal DNA or RNA transcripts, the target in studies to detect viral nucleic acids may represent double-or single-stranded (ds or ss) DNA or RNA (representative of viral genomes), nucleic acid repl ...

It has been possible for a number of years to detect RNA in situ in fixed cells or whole tissue-sections by the use of cloned probes (1–4). The probes are labeled either with radioactive nucleotides or modified nonradioactive nucleotides, such as biotinylated dUTP, which can be visualized by fluore ...

The technique of hybridization histochemistry (1) was developed as a means of locating specific mRNA populations in tissue sections using radiolabeled recombinant DNA probes with a complementary nucleotide sequence to the target mRNA. The original method used frozen sections fix ...

Quantitative in situ hybridization has emerged as a powerful technique for the study of gene expression within complex heterocellular systems, such as the central nervous system, by allowing the measurement of the relative levels of mRNA and their variations under experimental or phys ...

In basic and applied biomedical research there is a considerable interest in having reliable RNA in situ hybridization techniques, since these allow the detection of RNA expression in a morphological context at the individual cell level. Conventional molecular biology techniques f ...

The biochemical processes underlying development are ultimately dependent on accurately timed and regionally specific expression of particular genes. The latter fall into many categories, ranging from those specifying nuclear transcription factors through those coding ...

Hybridization histochemistry can be used at the light microscope level to determine the cellular site of gene expression in heterogeneous cell populations in tissue sections. However, the question often arises as to exactly which cell in a heterogeneous population is labeled and how this ...

Fluorescent in situ hybridization (FISH) is a powerful tool to analyze structural chromosome aberrations. The identification of structural abnormalities by routine and high resolution cytogenetic studies plays an important role in the diagnosis and treatment of disease. Howe ...

Chromosome painting refers to the complete decoration of specific metaphase chromosomes with complex probe mixtures. The painting signal is obtained by fluorescence in situ hybridization (FISH) of such mixtures established from a number of different sources (1–4). This technique ...

Fluorescence in situ hybridization (FISH) techniques are routinely used in physical mapping studies to determine the regional localization of gene and DNA sequences on human metaphase chromosomes (1). It is often difficult, however, to precisely position the hybridization signa ...

Alpha satellite DNA is a primate-specific family of tandemly repeated sequences present in the centromeric regions of all human chromosomes (1–3). The basic unit is a monomer repeat of approx 170 basepair (bp) that contains both sequences conserved among the different chromosomes and var ...

Telomeres and centromeres are critical structural and functional elements of eukaryotic chromosomes. Thus, they are important to our understanding of the organization and management of complex genomes. Termini of eukaryotic chromosomes are characterized by short, tandem rep ...

Yeast artificial chromosomes (YACs) (1) containing human inserts of up to 1 megabase (Mb) length have been mapped by fluorescence in situ hybridization (FISH) (for review see ref. 2). If total yeast clone DNA is used as a probe, an excess of yeast DNA (approx 98%) is labeled in addition to the human sequences (a ...

The yeast artificial chromosome (YAC) cloning system is capable of cloning large segments of DNA (50–2000 kb) from complex genomes (1). YAC genomic libraries have been constructed using DNA from human as well as other species and have been proven to be powerful tools for the analysis of the human geno ...

During the last two decades, techniques for the detection of specific DNA and RNA sequences in situ have been developed. In the beginning, only radioactive detection was possible, which to some extent made these techniques unsuited for routine purposes. Within the last decade, a number of nonra ...

The first part of this chapter (Sections 1.1 –1.6.) describes the complete positional cloning process with an emphasis on how multicolor in situ hybridization expedites these studies. Multicolor in situ hybridization is very useful in positional cloning because it maps cloned DNA quick ...

This chapter reviews data from in situ hybridization (ISH) experiments to determine the distribution, frequency, and intracellular localization of virus nucleic acids in tissue samples infected naturally with hepatitis viruses B, C, and D, and with human papillomaviruses. We have al ...