Department of Histopathology, Cambridge University Hospitals NHS Foundation Trust, Cambridge Biomedical Campus, Cambridge, UK.
Prostate. 2013 Jan;73(2):194-202. doi: 10.1002/pros.22556. Epub 2012 Jul 16.
The diagnosis and treatment of prostate cancer is a challenging global healthcare issue requiring significant molecular research. Such research frequently utilizes fresh frozen human tissue which needs to be obtained in a manner acceptable to the pathologist which does not compromise tumor diagnosis or staging.
Radical prostatectomy specimens were handled in a standardized method before being sliced fresh. Leaving the margins intact, multiple cylindrical cores were removed using a large skin punch and the sites were marked on a prostate map. The cylindrical cores were placed onto individual, pre-numbered foil squares and snap frozen in liquid nitrogen. Prostate maps were aligned with formalin-fixed paraffin embedded hematoxylin and eosin stained sections of the sampled slice to select tumor regions. Frozen tumor tissue cylinders were processed taking one section for hematoxylin and eosin staining, 6 µm × 50 µm sections for molecular studies and a further section for hematoxylin and eosin staining. This was performed for the length of the cylinder.
A total of 150 prostates have been removed and sliced using this technique. Pathological assessment remained uncompromised. Using the sequential hematoxylin and eosin stained frozen sections, cellularity could be monitored closely in tissues processed for research. The yield of RNA and DNA extracted was high (tumor mean 2.4 µg (RNA) and 12.7 ng per 300 µm tissue) and of high quality (mean tumor RIN 5.9).
This novel, rapid sampling and processing method provides high quality tissue for research without compromising pathology.
前列腺癌的诊断和治疗是一个具有挑战性的全球医疗保健问题,需要进行大量的分子研究。此类研究经常使用新鲜冷冻的人体组织,需要以病理学家可接受的方式获取,而不会影响肿瘤的诊断或分期。
在对新鲜组织切片之前,对根治性前列腺切除术标本进行了标准化处理。保留边缘完整,使用大皮肤冲孔取出多个圆柱形核心,并在前列腺图谱上标记部位。将圆柱形核心放在单独的、预先编号的箔正方形上,并在液氮中迅速冷冻。将前列腺图谱与取样切片的福尔马林固定石蜡包埋苏木精和伊红染色切片对齐,以选择肿瘤区域。对冷冻肿瘤组织圆柱进行处理,取一个用于苏木精和伊红染色的切片、6 µm × 50 µm 用于分子研究的切片和另一个用于苏木精和伊红染色的切片。这是在圆柱的长度上进行的。
共使用该技术切除并切片了 150 个前列腺。病理评估不受影响。使用连续的苏木精和伊红染色冷冻切片,可以密切监测用于研究的组织中的细胞活力。提取的 RNA 和 DNA 的产量很高(肿瘤平均 2.4 µg(RNA)和 300 µm 组织中 12.7 ng),质量也很高(肿瘤平均 RIN 5.9)。
这种新颖、快速的采样和处理方法为研究提供了高质量的组织,而不会影响病理学。