Stenersen T C, Danielsen H, Farrants G, Reith A
Department of Pathology, Norwegian Radium Hospital, Oslo.
Anal Cell Pathol. 1994 Oct;7(3):217-33.
Monolayer preparation from paraffin-embedded tissue from archival biopsy specimens older than 10 years was shown in this study to be more difficult to deal with than recent biopsies. We have studied the disseminative effect of several enzymes on archival biopsies from laryngeal epithelium, as well as mechanical dissemination. Thirty minutes incubation in protease 0.5% in 37 degrees C without syringing provided the best cell yield. Hydrolysis and Schiff staining conditions were also studied. Cell loss was calculated by stereological methods and found to be high in most cases. Cell loss during various steps in monolayer production was examined and we found no proof of selective cell loss. Large variations in Feulgen-Schiff staining intensities were observed, especially in biopsy specimens older than 10 years. The use of reliable internal (intrinsic) control cells is advocated since interspecimen variations in the diploid integrated optical density (IOD) can be expected. Although DNA measurements on archival biopsy material should be treated with caution, we demonstrate that meticulous studies of the preparation procedures will make it possible to obtain reliable histograms from old, archival biopsies.
本研究表明，从保存超过10年的存档活检标本的石蜡包埋组织制备单层细胞比近期活检标本更难处理。我们研究了几种酶对喉上皮存档活检标本的分散作用以及机械分散作用。在37℃下用0.5%蛋白酶孵育30分钟而不进行注射器冲洗可获得最佳细胞产量。还研究了水解和席夫染色条件。通过体视学方法计算细胞损失，发现大多数情况下细胞损失率很高。检查了单层细胞制备各个步骤中的细胞损失情况，未发现选择性细胞损失的证据。观察到福尔根-席夫染色强度存在很大差异，尤其是在保存超过10年的活检标本中。由于可以预期二倍体积分光密度（IOD）在不同标本间存在差异，因此提倡使用可靠的内部（固有）对照细胞。尽管对存档活检材料进行DNA测量时应谨慎，但我们证明，对制备程序进行细致研究将有可能从陈旧的存档活检标本中获得可靠的直方图。
