细胞功能测定

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 ...

During the process of characterizing the structure and function of novel antigens, it is usually necessary to create new monoclonal or polyclonal antibody reagents. However, once validated, these antibodies can be put to a multitude of experimental uses, such as detecting and quantitat ...

Culture of isolated microglia from dissociated cortical tissue has promoted the in vitro study of microglial function and morphological characteristics (Giulian and Baker, 1986; Streit and Kincaid-Colton, 1995). However, cultures prepared in this manner demonstrate charact ...

The purpose of this chapter is to introduce the reader to standard procedures for quantifying the number of neural cells in microcultures. The chick embryo was chosen as the source of neural cells, because embryos can be conveniently grown in an inexpensive laboratory incubator, in large, repl ...

This chapter focuses on the application of multiple-well plate technology to biological assays for neuroactive or other agents. It does not attempt to address specific cell types, culture systems, or individual assays for neurotoxicity, neural cell differentiation, or other applic ...

The ability to grow primary neurons under serum-free conditions is facilitating better control in studies of neuronal development, mechanisms of neuronal signaling, electrophysiology, pharmacology, plasticity, in vitro growth requirements, gene expression, and neuroto ...

Cell adhesion, cell contact, and intercellular interactions are critical to neuronal function. Neuronal cell surfaces are closely associated with the surfaces of other cells, including the specialized contacts of synapses, and the close appositions of astrocytes, oligodendro ...

Advances in molecular virology, and in understanding the molecular basis of human disease, has led to intensive efforts to use viruses for delivering therapeutic genes to cells. Although gene therapy (GT) to treat disease is still in its infancy, the techniques developed for gene delivery to c ...

The availability, in the past three decades, of well-characterized and immortalized neural cell lines has led to a rapid expansion of knowledge in many aspects of neurobiology. The major advantages of cell lines are that they are capable of long-term or indefinite growth, and generally repres ...