在多重复合成像中进行高效染色的最佳形式素固定的抛嵌入样品保存
Yuna Delarue1, Nadège Marec2, Marion Le Rochais1
1CHU de Brest, Brest, France; LBAI, UMR1227, Univ Brest, Inserm, Brest, France.
概括
室温储存会随着时间的推移降低免疫组织化学信号,影响诊断的准确性. 在-20°C或更低的温度下存储组织部分,可以保持信号完整性,以便进行可靠的分析.
科学领域:
- 生物病理学 生物病理学
- 免疫组织化学 免疫组织化学
- 生物标记分析 生物标记分析
背景情况:
- 免疫组织化学的准确性取决于组织样本的抗原完整性.
- 固定在甲中的胺嵌入式组织是常见的存储,但随着时间的推移信号降解是一个问题.
- 储存条件对标记物检测的影响仍然不太清楚.
研究的目的:
- 在存储的组织样本中调查信号降解的程度.
- 评估不同储存条件对标记物检测的影响.
- 评估标准化技术在维护数据质量的有效性.
主要方法:
- 在各种条件下 (室温,4°C,-20°C,-80°C) 保存长达8年的组织块和切片的分析.
- 使用25种针对免疫,树皮和结构标记物的抗体进行染色,使用成像质量细胞计 (IMC).
- 信号强度的量化和分析,通过imcRtools管道进行正常化或不正常化.
主要成果:
- 在室温存储块 (>6年) 中观察到逐渐的信号损失,影响特定标记.
- 在室温下几周内,对CD20,CD45和CD45RA等敏感标记物发生了显著的信号降解.
- -20°C或 -80°C的储存保持了染色质量;4°C提供了部分保存.
- 正常化部分纠正了变化,但无法恢复严重退化的信号.
结论:
- 室温储存显著降低了IMC检测到的免疫活性,可能影响科学解释和诊断.
- 在-20°C下存储组织部分是一种有效且易于使用的方法,可以保持免疫组织化学质量.
- 对于回顾性研究,优先考虑存储不到六年的组织块;标准化的保存协议对于可重复性至关重要.
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