Mathieson William, Marcon Nathalie, Antunes Laurent, Ashford David A, Betsou Fay, Frasquilho Sonia G, Kofanova Olga A, McKay Siobhan C, Pericleous Stephan, Smith Colleen, Unger Kristian M, Zeller Constanze, Thomas Geraldine A
From the Integrated Biobank of Luxembourg, Luxembourg Department of Surgery and Cancer, Imperial College London, London, United Kingdom.
From the Integrated Biobank of Luxembourg, Luxembourg.
Am J Clin Pathol. 2016 Jul;146(1):25-40. doi: 10.1093/ajcp/aqw023.
To evaluate the PAXgene tissue fixation system.
Clinical biospecimens (n = 46) were divided into PAXgene-fixed paraffin-embedded (PFPE), formalin-fixed paraffin-embedded (FFPE), and fresh-frozen (FF) blocks. PFPE and FFPE sections were compared for histology (H&E staining) and immunohistochemistry (14 antibodies) using tissue microarrays. PFPE, FFPE, and FF samples were compared in terms of RNA quality (RNA integrity number, polymerase chain reaction [PCR] amplicon length, and quantitative reverse transcription PCR), DNA quality (gel electrophoresis and methylation profiling) and protein quality (liquid chromatography-mass spectrometry [LC-MS/MS]).
PFPE protocol optimization was required in most cases and is described. RNA extracted from PFPE sections was considerably less degraded than that from FFPE sections but more degraded than that from FF blocks. Genomic-length DNA was extracted from PFPE and FF biospecimens, and methylation profiling showed PFPE and FF biospecimens to be almost indistinguishable. Only degraded DNA was extracted from FFPE biospecimens. PFPE sections yielded peptides that were slightly less amenable to LC-MS/MS analysis than FFPE sections, but FF gave slightly better results.
While it cannot be envisaged that PAXgene will replace formalin in a routine clinical setting, for specific projects or immunodiagnostics involving biospecimens destined for immunohistochemical or histologic staining and DNA or RNA analyses, PAXgene is a viable option.
评估PAXgene组织固定系统。
将临床生物样本（n = 46）分为PAXgene固定石蜡包埋（PFPE）、福尔马林固定石蜡包埋（FFPE）和新鲜冷冻（FF）样本块。使用组织芯片比较PFPE和FFPE切片的组织学（苏木精-伊红染色）和免疫组织化学（14种抗体）。比较PFPE、FFPE和FF样本的RNA质量（RNA完整性数值、聚合酶链反应[PCR]扩增子长度和定量逆转录PCR）、DNA质量（凝胶电泳和甲基化分析）和蛋白质质量（液相色谱-质谱联用[LC-MS/MS]）。
大多数情况下需要对PFPE方案进行优化，并对此进行了描述。从PFPE切片中提取的RNA降解程度明显低于FFPE切片，但高于FF样本块。从PFPE和FF生物样本中提取了基因组长度的DNA，甲基化分析显示PFPE和FF生物样本几乎无法区分。仅从FFPE生物样本中提取到了降解的DNA。PFPE切片产生的肽段对LC-MS/MS分析的适用性略低于FFPE切片，但FF的结果略好。
虽然无法设想PAXgene会在常规临床环境中取代福尔马林，但对于涉及用于免疫组织化学或组织学染色以及DNA或RNA分析的生物样本的特定项目或免疫诊断，PAXgene是一个可行的选择。
