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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Page: All 1 2 3 4 5 6 Related biology technology :1.
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