( ...)The DIG System Nonradioactive Automated,,,High-Throughput In Situ Hybridization:,,,a Powerful Tool for Functional Genomics Research

Over the past decade, many organisms have been the subject of large-scale genome projects, and as a result a tremendous number of gene sequences are now ready for functional analysis. Knowledge of tissues and cells that express particular genes is key to understanding gene function.

Microarrays and similar high-throughput gene expression technologies are well established and widely used to determine the expression of large numbers of genes in parallel. Their cellular resolution and accuracy of expression patterns, however, are limited to the level of tissue dissection. Nonetheless, they are excellent filters for the selection of genes that are suitable for analysis by in situ hybridization.

In situ hybridization is a gene expression technique that provides spatial detail and allows the detection of very small numbers of positive cells in an intact tissue context. The technique is vital for the functional analysis of genes, and it has been used extensively in biological and medical research for more than 20 years. To allow the simultaneous analysis of large numbers of genes by in situ hybridization, the procedure was automated fairly recently.

Automation of nonradioactive in situ hybridization not only increases throughput, but also overcomes its major impediments, i.e., that it is technically challenging, laborintensive, and prone to human error, by exerting accurate control of critical parameters such as temperature, pipetting volume, incubation time, and number of repetitions.

There are only a few instruments for automated in situ hybridization available on the market. For very small, permeable tissue samples such as animal eggs, small l
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