Automation of fluorescence in situ hybridization pretreatment: A comparative study of different sample types

Background: In fluorescence in situ hybridization (FISH) applications, the efficiency of probe hybridization is greatly enhanced by treating the cell or tissue preparation with a variety of reagents that make the target perme able while preserving important morphological features. Pretreatment protoc ols can be very labor intensive, adding cost to the test. The automation of specimen pretreatment eliminates human variation in the procedure through standardization of such variables as treatment times and temperatures. Methods and Results: We developed an instrument, the VP 2000 Processor, tha t can process up to 50 specimens simultaneously under computer control. In this comparative FISH study of matched specimens, one set was processed acc ording to the manual pretreatment protocol and compared with specimens proc essed by the VP 2000. Processed specimen types included paraffin-embedded b reast tissue, uncultured amniocytes, bone marrow, peripheral-blood lymphocy tes, and uroepithelial cells recovered from urine. Data show equivalent or brighter and more specific overall signal quality compared with matched man ually executed controls. Conclusions: The automation of sample pretreatment for FISH provides a supe rior, more consistent level of performance than the manual format.