Development of full-automatic in situ hybridization system for Diagnosis

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We first tested the availability of commercially available "Full automatic histological staining system" and "Full automatic immunostaining system for in situ hybridization system. The combination of both systems did work considarably well in not only to detect viral DNA but mRNAs. However, we recognized that some parts of the instrument need to much more heat resistant for long term use. So we changed some of the parts to the goods resistant to high temperature. Next we investigated the TSA, the amplifier to detect signal. We confirmed the detection limit of the signal raised to approximately 50 times than usual system. This amplifier would be useful for obtaining stable result of in situ hybridization. Finally we tested if we can detect gene amplified product of oncogenes such as ras and sam-1 from sample obtained operation. As a result, we detected amplified sam-1 gene at the efficiency of 30% in the sample. This value was consistent from that obtained by Southern blotting. However, it was difficult to detect amplified gene from pathological samples which were already fixed by formaline. This may due to the regradation of DNA during fixiation. We still need to search optical condition to detect nucleic acid from pathological samples.