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目的:探讨在FISH实验中,乳腺癌石蜡组织切片的最适酶的消化时间。方法:收集42例手术切除的乳腺癌标本,经过固定、包埋和切片,其中12份标本的组织切片进行胃蛋白酶消化时间的优化实验,分别观察酶的消化时间为10分钟,15分钟,20分钟和25分钟对FISH结果的影响,随后在酶的优化条件对剩余30份组织切片继续杂交实验。结果:酶消化10min提示消化时间不足,酶消化15min提示消化时间适合,结果能很好地判读,而酶消化25min提示消化过度。对剩余的30份组织切片进行了18min的消化,杂交信号基本都能判读。结论:本实验条件下,石蜡包埋切片的最适消化时间在15-20min,不同实验室及不同的FISH处理过程会略有差异。所以建议FISH实验之前,应对酶的消化时间首先进行摸索。
Objective: To investigate the optimal enzymatic digestion time of paraffin tissue sections of breast cancer in FISH experiment. Methods: Forty-two specimens of surgically resected breast cancer were collected, fixed, embedded and sectioned. Tissue sections of 12 specimens were optimized for pepsin digestion time. The digestion time was 10 minutes, 15 minutes and 20 minutes respectively Minutes and 25 minutes on the FISH results, followed by the hybridization experiments on the remaining 30 tissue sections under the optimized conditions of the enzyme. Results: Enzyme digestion for 10 minutes showed that the digestion time was insufficient. Enzyme digestion for 15 minutes suggested that the digestion time was suitable and the result was well interpreted. Enzyme digestion for 25 minutes suggested over-digestion. The remaining 30 tissue sections were digested for 18 min, the hybridization signal can be interpreted basically. CONCLUSION: Under the conditions of this experiment, the optimum digestion time of paraffin embedded sections is 15-20min, which will be slightly different in different laboratories and different FISH treatments. Therefore, it is recommended that FISH experiments should be conducted first to deal with the digestion time of the enzyme.