Tan Leonard Hwan Cheong, Do Elaine, Chong Siew Meng, Koay Evelyn Siew Chuan
Department of Pathology, National University of Singapore, Singapore.
Mol Diagn. 2003;7(1):27-33. doi: 10.1007/BF03260017.
It is widely known that the efficiency of fluorescence in situ hybridization (FISH) probes applied to formalin-fixed, paraffin-embedded tissues is affected by the conditions under which the tissues are fixed and embedded. However, relatively few studies address exactly how tissue archiving conditions affect the performance of FISH probes. We report our experience based on use of an ALK FISH probe, during the validation of its diagnostic utility.
We applied the probe to 77 formalin-fixed, paraffin-embedded tissue blocks archived from 1991 through to 2000, and studied the interrelationship between the archival age (which ranged up to 10 years), type and condition of tissue, duration required for optimum hydrolysis, and obtainability of hybridization signals.
We found that as archival age and tissue collagen content increased, not only did hydrolysis times have to be prolonged in order to yield interpretable hybridization signals, but also the likelihood of blocks becoming non-signaling increased. The most striking positive correlations were seen between the archival age of signaling lymphoid blocks and their requisite hydrolysis times.
The difficulty in applying FISH on archival tissue increases with its archival age and collagen content, and may necessitate changes in laboratory protocol accordingly.
众所周知，应用于福尔马林固定、石蜡包埋组织的荧光原位杂交（FISH）探针的效率受组织固定和包埋条件的影响。然而，相对较少的研究确切探讨组织存档条件如何影响FISH探针的性能。我们报告在验证ALK FISH探针诊断效用期间基于其使用的经验。
我们将该探针应用于1991年至2000年存档的77个福尔马林固定、石蜡包埋组织块，研究存档年限（最长达10年）、组织类型和状况、最佳水解所需时间以及杂交信号的可获得性之间的相互关系。
我们发现，随着存档年限和组织胶原含量增加，不仅为产生可解读的杂交信号必须延长水解时间，而且组织块无信号的可能性也增加。在有信号的淋巴组织块的存档年限与其所需水解时间之间观察到最显著的正相关。
对存档组织应用FISH的难度随其存档年限和胶原含量增加而增加，可能需要相应改变实验室方案。
