储存组织块中原位杂交检测RNA的快速丢失及未染色玻片冷藏保存法
Rapid Loss of RNA Detection by In Situ Hybridization in Stored Tissue Blocks and Preservation by Cold Storage of Unstained Slides.
作者信息
Baena-Del Valle Javier A, Zheng Qizhi, Hicks Jessica L, Fedor Helen, Trock Bruce J, Morrissey Colm, Corey Eva, Cornish Toby C, Sfanos Karen S, De Marzo Angelo M
机构信息
Department of Pathology.
Department of Pathology and Laboratory Medicine, Fundacion Santa Fe de Bogota University Hospital, Bogota DC, Colombia.
出版信息
Am J Clin Pathol. 2017 Nov 2;148(5):398-415. doi: 10.1093/ajcp/aqx094.
OBJECTIVES
Recent commercialization of methods for in situ hybridization using Z-pair probe/branched DNA amplification has led to increasing adoption of this technology for interrogating RNA expression in formalin-fixed, paraffin-embedded (FFPE) tissues. Current practice for FFPE block storage is to maintain them at room temperature, often for many years.
METHODS
To examine the effects of block storage time on FFPE tissues using a number of RNA in situ probes with the Advanced Cellular Diagnostic's RNAscope assay.
RESULTS
We report marked reductions in signals after 5 years and significant reductions often after 1 year. Furthermore, storing unstained slides cut from recent cases (<1 year old) at -20°C can preserve hybridization signals significantly better than storing the blocks at room temperature and cutting the slides fresh when needed.
CONCLUSIONS
We submit that the standard practice of storing FFPE tissue blocks at room temperature should be reevaluated to better preserve RNA for in situ hybridization.
目的
使用Z对探针/分支DNA扩增的原位杂交方法最近实现商业化，导致该技术越来越多地被用于检测福尔马林固定、石蜡包埋（FFPE）组织中的RNA表达。目前FFPE组织块的保存方法是将它们保存在室温下，通常保存多年。
方法
使用先进细胞诊断公司的RNAscope检测法，用多种RNA原位探针检测组织块保存时间对FFPE组织的影响。
结果
我们报告称，5年后信号显著减少，1年后通常也会显著减少。此外，将近期病例（<1年）切出的未染色玻片保存在-20°C，比将组织块保存在室温下并在需要时新鲜切片能更好地保存杂交信号。
结论
我们认为，应重新评估将FFPE组织块保存在室温下的标准做法，以便更好地保存用于原位杂交的RNA。
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