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固定时间对人福尔马林固定石蜡包埋脑组织中信使核糖核酸可检测性的影响。

Impact of fixation duration on messenger RNA detectability in human formalin-fixed paraffin-embedded brain tissue.

作者信息

Hurler Charlene-Annett, Liebscher Sabine, Arzberger Thomas, Jäkel Sarah

机构信息

Institute for Stroke and Dementia Research, LMU University Hospital, Ludwig-Maximilians-University Munich, 81377 Munich, Germany.

Institute of Clinical Neuroimmunology, LMU University Hospital, Ludwig-Maximilians-University Munich, 82152 Planegg-Martinsried, Germany.

出版信息

Brain Commun. 2024 Nov 28;6(6):fcae430. doi: 10.1093/braincomms/fcae430. eCollection 2024.

DOI:10.1093/braincomms/fcae430
PMID:39659968
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11630792/
Abstract

Technologies to study mRNA in post-mortem human brain samples have greatly advanced our understanding of brain pathologies. With ongoing improvements, particularly in formalin-fixed paraffin-embedded tissue, these technologies will continue to enhance our knowledge in the future. Despite various considerations for tissue and mRNA quality, such as pre-mortem health status and RNA integrity, the impact of the tissue fixation time has not been addressed in a systemic fashion yet. In this study, we employed RNAscope to assess mRNA detectability in human post-mortem brain tissue in relation to fixation time. Our results reveal a dynamic change in mRNA detection across varying fixation durations, accompanied by an increase in signal derived from the negative probe and autofluorescence background. These findings highlight the critical relevance of standardized fixation protocols for the collection of human brain tissue in order to probe mRNA abundancy to ensure reliable and comparable results.

摘要

用于研究死后人类大脑样本中mRNA的技术极大地推进了我们对脑部病理学的理解。随着技术的不断改进，特别是在福尔马林固定石蜡包埋组织方面，这些技术在未来将继续增进我们的知识。尽管对组织和mRNA质量有各种考量因素，如生前健康状况和RNA完整性，但组织固定时间的影响尚未得到系统的研究。在本研究中，我们使用RNAscope评估人类死后脑组织中mRNA可检测性与固定时间的关系。我们的结果揭示了在不同固定持续时间内mRNA检测的动态变化，同时伴随着来自阴性探针的信号和自发荧光背景的增加。这些发现凸显了标准化固定方案对于收集人类脑组织以探测mRNA丰度的关键相关性，从而确保获得可靠且可比的结果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34f8/11630792/2818a3591600/fcae430f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34f8/11630792/e50342d12de7/fcae430_ga.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34f8/11630792/fafeb502ad1b/fcae430f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34f8/11630792/f5cddd7af3e7/fcae430f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34f8/11630792/2818a3591600/fcae430f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34f8/11630792/e50342d12de7/fcae430_ga.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34f8/11630792/fafeb502ad1b/fcae430f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34f8/11630792/f5cddd7af3e7/fcae430f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34f8/11630792/2818a3591600/fcae430f3.jpg

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本文引用的文献

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Single-Nucleus RNA-Seq: Open the Era of Great Navigation for FFPE Tissue.
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Single-cell and spatial transcriptomics: deciphering brain complexity in health and disease.单细胞和空间转录组学：解析健康和疾病中的大脑复杂性。
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Selective time-dependent changes in activity and cell-specific gene expression in human postmortem brain.人类尸检脑中活性和细胞特异性基因表达的选择性时变。
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