气候变化领域集成服务门户(CCPortal): IBEX: A versatile multiplex optical imaging approach for deep phenotyping and spatial analysis of cells in complex tissues

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DOI	10.1073/PNAS.2018488117
	IBEX: A versatile multiplex optical imaging approach for deep phenotyping and spatial analysis of cells in complex tissues
	Radtke A.J.; Kandov E.; Lowekamp B.; Speranza E.; Chu C.J.; Gola A.; Thakur N.; Shih R.; Yao L.; Yaniv Z.R.; Beuschel R.T.; Kabat J.; Croteau J.; Davis J.; Hernandez J.M.; Germain R.N.
发表日期	2021
ISSN	00278424
起始页码	33455
结束页码	33465
卷号	117 期号:52
英文摘要	The diverse composition of mammalian tissues poses challenges for understanding the cell–cell interactions required for organ homeostasis and how spatial relationships are perturbed during disease. Existing methods such as single-cell genomics, lacking a spatial context, and traditional immunofluorescence, capturing only two to six molecular features, cannot resolve these issues. Imaging technologies have been developed to address these problems, but each possesses limitations that constrain widespread use. Here we report a method that overcomes major impediments to highly multiplex tissue imaging. “Iterative bleaching extends multiplexity” (IBEX) uses an iterative staining and chemical bleaching method to enable high-resolution imaging of >65 parameters in the same tissue section without physical degradation. IBEX can be employed with various types of conventional microscopes and permits use of both commercially available and user-generated antibodies in an “open” system to allow easy adjustment of staining panels based on ongoing marker discovery efforts. We show how IBEX can also be used with amplified staining methods for imaging strongly fixed tissues with limited epitope retention and with oligonucleotide-based staining, allowing potential cross-referencing between flow cytometry, cellular indexing of transcriptomes and epitopes by sequencing, and IBEX analysis of the same tissue. To facilitate data processing, we provide an open-source platform for automated registration of iterative images. IBEX thus represents a technology that can be rapidly integrated into most current laboratory workflows to achieve high-content imaging to reveal the complex cellular landscape of diverse organs and tissues. © 2020 National Academy of Sciences. All rights reserved.
英文关键词	High-dimensional imaging; Immune system; Immunofluorescence; Quantitative microscopy; Tissue immunity
语种	英语
scopus关键词	epitope; transcriptome; fluorescent dye; animal experiment; animal tissue; Article; bleaching; cell interaction; controlled study; cytology; flow cytometry; fluorescence imaging; histology; human; human tissue; image analysis; image registration; immune system; immunofluorescence; iterative bleaching extends multiplexity; mouse; nonhuman; phenotype; priority journal; sequence analysis; spatial analysis; staining; transcriptomics; workflow; animal; antibody specificity; cells; diagnostic imaging; fluorescence imaging; image processing; immunization; lymph node; metabolism; phenotype; procedures; Animals; Cells; Fluorescent Dyes; Humans; Image Processing, Computer-Assisted; Immunization; Lymph Nodes; Mice; Optical Imaging; Organ Specificity; Phenotype
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/179678
作者单位	Center for Advanced Tissue Imaging, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, United States; Lymphocyte Biology Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, United States; Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, TN 37240, United States; Bioinformatics and Computational Bioscience Branch, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD 20892, United States; Innate Immunity and Pathogenesis Section, Laboratory of Virology, National Institute of Allergy and Infectious Diseases, NIH, Hamilton, MT 59840, United States; Rockefeller University, New York, NY 10065, United States; Department of Science Education, Howard Hughes Medical Institute, Chevy Chase, MD 20815, United States; Biological Imaging Section, Research Technologies Branch, National Institute of Allergy and Infectious Disea...
推荐引用方式 GB/T 7714	Radtke A.J.,Kandov E.,Lowekamp B.,et al. IBEX: A versatile multiplex optical imaging approach for deep phenotyping and spatial analysis of cells in complex tissues[J],2021,117(52).
APA	Radtke A.J..,Kandov E..,Lowekamp B..,Speranza E..,Chu C.J..,...&Germain R.N..(2021).IBEX: A versatile multiplex optical imaging approach for deep phenotyping and spatial analysis of cells in complex tissues.Proceedings of the National Academy of Sciences of the United States of America,117(52).
MLA	Radtke A.J.,et al."IBEX: A versatile multiplex optical imaging approach for deep phenotyping and spatial analysis of cells in complex tissues".Proceedings of the National Academy of Sciences of the United States of America 117.52(2021).