多重离子束成像飞行时间(MIBI-TOF)在存档人类组织中的可重现、高维成像。

Reproducible, high-dimensional imaging in archival human tissue by multiplexed ion beam imaging by time-of-flight (MIBI-TOF).

机构信息

Department of Pathology, Stanford University, Stanford, CA, 94304, USA.

Office of the Director, Division of Cancer Treatment and Diagnosis, National Cancer Institute, Bethesda, MD, 20892, USA.

出版信息

Lab Invest. 2022 Jul;102(7):762-770. doi: 10.1038/s41374-022-00778-8. Epub 2022 Mar 29.

Abstract

Multiplexed ion beam imaging by time-of-flight (MIBI-TOF) is a form of mass spectrometry imaging that uses metal labeled antibodies and secondary ion mass spectrometry to image dozens of proteins simultaneously in the same tissue section. Working with the National Cancer Institute's (NCI) Cancer Immune Monitoring and Analysis Centers (CIMAC), we undertook a validation study, assessing concordance across a dozen serial sections of a tissue microarray of 21 samples that were independently processed and imaged by MIBI-TOF or single-plex immunohistochemistry (IHC) over 12 days. Pixel-level features were highly concordant across all 16 targets assessed in both staining intensity (R = 0.94 ± 0.04) and frequency (R = 0.95 ± 0.04). Comparison to digitized, single-plex IHC on adjacent serial sections revealed similar concordance (R = 0.85 ± 0.08) as well. Lastly, automated segmentation and clustering of eight cell populations found that cell frequencies between serial sections yielded an average correlation of R = 0.94 ± 0.05. Taken together, we demonstrate that MIBI-TOF, with well-vetted reagents and automated analysis, can generate consistent and quantitative annotations of clinically relevant cell states in archival human tissue, and more broadly, present a scalable framework for benchmarking multiplexed IHC approaches.

摘要

多重离子束成像飞行时间(MIBI-TOF)是一种质谱成像技术,它使用金属标记的抗体和二次离子质谱,在同一块组织切片中同时对数十种蛋白质进行成像。我们与美国国立癌症研究所(NCI)的癌症免疫监测和分析中心(CIMAC)合作,进行了一项验证研究,评估了 21 个样本的组织微阵列的 12 个连续切片之间的一致性,这些样本是由 MIBI-TOF 或单重免疫组化(IHC)独立处理和成像的,历时 12 天。在对所有 16 个目标进行的像素级特征评估中,染色强度(R=0.94±0.04)和频率(R=0.95±0.04)的一致性都很高。与相邻连续切片上的数字化单重 IHC 进行比较,也显示出相似的一致性(R=0.85±0.08)。最后,对 8 种细胞群的自动分割和聚类表明,连续切片之间的细胞频率平均相关性为 R=0.94±0.05。总之,我们证明了 MIBI-TOF 可以与经过充分验证的试剂和自动化分析相结合,对存档的人类组织中的临床相关细胞状态进行一致和定量的注释,并且更广泛地为多重 IHC 方法的基准测试提供了一个可扩展的框架。

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