Heat-induced antigen retrieval in fluorescence in situ hybridization: An effective approach enhancing signal intensity in poor-quality FFPE sections.
Heat-induced antigen retrieval in fluorescence in situ hybridization: An effective approach enhancing signal intensity in poor-quality FFPE sections.
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DOI：
10.3892/etm.2021.10915
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发表时间：
2021-12
影响因子：
2.7
通讯作者：
Li L
中科院分区：
文献类型：
--
作者：
Yu Q;Zhang C;Huang W;Li L
荧光原位杂交（FISH）作为评估染色体异常的辅助工具，在鉴别诊断和治疗决策中很重要。在临床实践中，病理学家经常会遇到福尔马林固定石蜡包埋（FFPE）切片，荧光信号弱，效果不理想，主要是由于组织处理或保存不当，导致解释困难。在本研究中，收集了常规FISH无法检测的FFPE样品。使用柠檬酸缓冲液或Tris-EDTA缓冲液引入热诱导抗原检索（HIAR），而不是使用商业试剂盒的预处理步骤，而随后的实验工作流程保持不变。经过hiar辅助FISH后，杂交效率和信号强度明显增强，两种AR溶液效果比较，信号充分性无差异。本研究表明，HIAR是FISH的可靠工具，特别是对于传统分析中荧光信号微弱或没有荧光信号的低质量FFPE切片。
Fluorescence in situ hybridization (FISH) serves as an ancillary tool for assessing chromosomal abnormalities and is important in differential diagnoses and treatment decisions. In clinical practice, pathologists encounter unsatisfactory formalin-fixed paraffin-embedded (FFPE) sections exhibiting weak fluorescence signals, mostly due to inappropriate tissue processing or preservation, leading to interpretation difficulties. For the present study, FFPE samples for which conventional FISH failed were collected. Instead of a pretreatment step using a commercial kit, heat-induced antigen retrieval (HIAR) was introduced using either citrate buffer or Tris-EDTA buffer, while the subsequent experimental workflow remained unchanged. After HIAR-assisted FISH, the hybridization efficiency and signal intensity were markedly enhanced and no difference in signal adequacy was observed when comparing the effect of the two AR solutions. The present study demonstrated that HIAR is a reliable tool for FISH, particularly for poor-quality FFPE sections yielding weak or no fluorescence signals in the conventional analysis.
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