细胞技术

When combined with molecular analysis and immunocytological localization, fluorescence in situ hybridization (FISH) represents one of the most direct and precise experimental tools in current biological research (1-4). Direct visualization and in situ detection fill the gap b ...

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 ...

We have had a long term interest in the neuropathogenesis of human immunodeficiency virus (HIV)-associated neurological diseases. Neurological disease is a prominent feature of HIV-1 infection, usually occurring during the late stages of acquired immunodeficiency syndrome ( ...

The successful implementation of intracytoplasmic sperm injection (ICSI) has provided a unique means to allow couples suffering from severe male infertility to achieve their reproductive goals. If the infertile man has a germ cell in the ejaculate or retrievable from the reproductive ...

Whole chromosome painting libraries (WCPLs) have provided a very powerful tool to cytogeneticists. The technique allows the painting of specific chromosomes in metaphase spreads and in interphase nuclei (1-4). The usefulness of WCPLs is particularly evident in identifying the chr ...

Since the first description of the technique of in situ hybridization to detect specific mRNA species on tissue sections, a variety of methods have been employed. Preparation of tissue and cells ranges from the use of frozen sections, which show good preservation of mRNA but poor morphology, to st ...

Although largely replaced by the use of fluorescent in situ hybridization (FISH) in animal and human molecular cytogenetics, the technique of radioactive in situ hybridization (RISH) still has some uses. Using practicable exposure times for autoradiographs of 3–4 wk, RISH is approx 50 tim ...

Since the early analyses of gene expression in the Drosophila embryo (1), whole-mount in situ hybridization has become one of the most powerful and versatile tools in developmental biology. The ability to visualize a gene’s expression both in time and space is a necessary first step in investiga ...

In the great majority of cases in situ hybridization is used to localize mRNA species at the tissue level, or DNA at the chromosome level. These approaches are generally best done by light microscopy. There are instances, however, when it becomes important to localize nucleic acids at the subcellu ...

Detection of nucleic acid sequences at the ultrastructural level allows the localization of RNA and DNA molecules to specific subcellular compartments. At the light microscope level, it is already evident that DNA and RNA sequences do not have a random distribution, suggesting that their ...

In situ hybridization is the only DNA- or RNA-based molecular biology based test that allows for the direct correlation of the results with the histologic and cytologic features of the sample. The DNA/RNA extraction that precedes filter hybridization (slot blot, hybrid capture, or Southern ...

In situ hybridization (ISH) permits the localization of specific unique or repeated DNA and RNA sequences at the level of individual cells (1-4). It has significantly advanced the study of gene structure and expression, and, in addition to morphological identification of cell types involv ...

in situ hybridization can provide accurate intracellular localization of specific viral nucleic acids in infected tissues and cells. The technique, although conceptually simple, is affected by many variables including: stability and accessibility of target sequences; meth ...

Sensitive techniques developed to detect biologically relevant proteins at the mRNA and protein levels have been major research tools in basic and applied biomedical research (1-6). The combination of these two techniques has been particularly valuable when the biological protein ...

Apoptosis is a morphologically distinct form of programmed cell death. It has a role in such processes as embryogenesis, immune regulation, and defense against viruses, and can also be induced by a variety of physical and chemical stimuli (1). Importantly apoptosis leads to the safe removal of ce ...

Although fluorescent in situ hybridization (FISH) technology has been used extensively for gene mapping and genome analysis (1-8), methods that visualize the interaction of DNA and protein are required to elucidate the functional aspects of the chromosome. By displaying the physical ...

For several years, quantitative in situ hybridization has emerged as a particularly powerful technique to study gene expression within complex heterocellular systems, including the central nervous system, by measuring mRNA levels and their variations under experimental or ph ...

In situ hybridization takes advantage of the reaction between two complementary single-stranded nucleic acid molecules. These molecules bind by means of hydrogen bonding of complementary base pairs. Hybridization performed in situ includes techniques that detect these hybr ...

Comparative genome analysis between two distantly related species allows the organization of genes to be traced from a common ancestor. When several genes are mapped in one species and these genes are then localized in the distantly related species, then the genomic content of this region can be ...

Since its development by Pardue et al. (1), the technique of in situ hybridization to polytene chromosomes has played a central role in the molecular genetic analysis of Drosophila melanogaster. The power of in situ hybridization is due largely to the scale of polytene chromosomes and conseque ...