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

d coverslipped with aqueous mounting medium.



For automated nonradioactive in situ hybridization, the Tecan Freedom EVO robot 150/8 pipetting instrument (eight liquid-dispensing needles) with GenePaint system components (Figure 1) is used.

Microscopy

Slides generated by the in situ hybridization robot are coverslipped and photographed in a compound microscope Leica DMR microscope, a motorized Mrzhuser stage that accommodates up to eight slides, a Leica electronic focusing system, a JVC CCD camera, and a PC-based controller that drives stage and camera. A detailed version of the procedure can be obtained from the author.

Results

Using the DIG system for nonradioactive in situ hybridization in combination with the Tecan pipetting instrument, Georg Eicheles group has successfully adapted in situ hybridization to high-throughput and routinely obtains excellent and consistent results (Figures 2 and 3).



The expression patterns are highly specific and reproducible with a high signal-to-noise ratio. Using a set of 20 slides representing different tissues or developmental stages per gene, 50 genes can be analyzed per week if 200 slide positions are available (Figure 1b, c) [1].



Summary and Conclusion

The DIG system has been applied successfully to highthroughput, automated in situ hybridization and gives excellent and consistent results, allowing the routine analysis of a large number of probes. By automation of nonradioactive in situ hybridization and adaptation for high-throug
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