Characterizing and Diminishing Autofluorescence in Formalin-fixed Paraffin-embedded Human Respiratory Tissue
Characterizing and Diminishing Autofluorescence in Formalin-fixed Paraffin-embedded Human Respiratory Tissue
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DOI：
10.1369/0022155414531549
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发表时间：
2014-06-01
影响因子：
3.2
通讯作者：
Taubenberger, Jeffery K.
中科院分区：
文献类型：
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作者：
Davis, A. Sally;Richter, Anke;Taubenberger, Jeffery K.
组织自体荧光经常妨碍福尔马林固定石蜡包埋的呼吸道组织中免疫荧光标记物的可视化。我们评估了9种治疗方法，据报道，这些治疗方法在减少其他组织类型的自体荧光方面有效。三种最有效的是铬黑T、苏丹黑B和硼氢化钠，如使用白色光激光共聚焦（2）（多λ）分析所测量的。我们还评估了蒸汽抗原修复和血清应用对人气管组织自体荧光的影响。将该（2）数据与2维高斯表面进行功能拟合，结果显示蒸汽抗原修复和血清应用对自体荧光的贡献最小，并且三种治疗方法均有效。总之，这些研究为减少福尔马林固定石蜡包埋的人呼吸道组织中的自发荧光提供了一套指导方针。此外，这些表征技术可转移到其他组织类型中的类似问题，如在不同固定剂中固定的冷冻人肝组织和石蜡包埋的小鼠肺组织上所证明的。
Tissue autofluorescence frequently hampers visualization of immunofluorescent markers in formalin-fixed paraffin-embedded respiratory tissues. We assessed nine treatments reported to have efficacy in reducing autofluorescence in other tissue types. The three most efficacious were Eriochrome black T, Sudan black B and sodium borohydride, as measured using white light laser confocal (2) (multi-lambda) analysis. We also assessed the impact of steam antigen retrieval and serum application on human tracheal tissue autofluorescence. Functionally fitting this (2) data to 2-dimensional Gaussian surfaces revealed that steam antigen retrieval and serum application contribute minimally to autofluorescence and that the three treatments are disparately efficacious. Together, these studies provide a set of guidelines for diminishing autofluorescence in formalin-fixed paraffin-embedded human respiratory tissue. Additionally, these characterization techniques are transferable to similar questions in other tissue types, as demonstrated on frozen human liver tissue and paraffin-embedded mouse lung tissue fixed in different fixatives.