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The areas of invading carcinoma cells in each specimen were analysed statistically by using BZ analyser (KEYENCE, Osaka, Japan).
At least five metaphases per specimen were analysed.
The specimen were analysed by confocal microscopy (Zeiss, LSM 510 Meta).
At least ten metaphases per specimen were analysed, in some cases sequentially.
At least 30 tumour cells from each specimen were analysed if the sample was deemed homogeneous, but at least 60 cells were analysed if the sample was found to contain between 5 and 10 HER2 gene copies in >50% of the area selected for evaluation.
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Furthermore, a sequential follow-up specimen was analysed from one patient undergoing adenomatous polyp review.
Usually, the specimen was analysed with haematoxylin and eosin stain and other stains as required.
However, each specimen was analysed in great depth, including 24 cut sections and 47 histological slides.
Though a strict application of sequential sampling would require a single additional specimen be analysed if required, it would be more practical to increment by five.
The RNA quality in each clinical specimen was analysed by RT PCR of 28S rRNA using 25 ng of total RNA and 35 cycles of amplification.
The RNA quality in each clinical specimen was analysed by RT PCR of 28S rRNA using 25 ng of total RNA and 35 amplification cycles.
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