ArticleNature protocols2022
IBEX: an iterative immunolabeling and chemical bleaching method for high-content imaging of diverse tissues.
Article in Nature protocols, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 98 papers.
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Who cites it
98 citing papers in PubMed, 136 citations in OpenAlex.
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- Single-Cell CRISPR: An Efficient Strategy for Decoding Plant Cis-Regulatory Complexity.Plant biotechnology journal · 2026Review
- Advances in spatial omics for the analysis of prostate cancer.Analytical and bioanalytical chemistry · 2026Review
- Image-based, pooled phenotyping reveals multidimensional, disease-specific variant effects.Cell · 2026Article
- CD4Science (New York, N.Y.) · 2026Article
- Image Tracing of Inflammatory Intestinal Organoids via Computational Clearing.Nanomaterials (Basel, Switzerland) · 2026Article
- Microglia-independent rAAV-induced inflammation causes persistent ocular immune dysregulation rescued by S1P receptor modulation.Molecular therapy : the journal of the American Society of Gene Therapy · 2026Article
- IL10R Inhibition Induces Neutrophil Tumoricidal Activity.Cancer immunology research · 2026Article
- Spatial proteomic mapping of the human and mouse retina using IBEX.JCI insight · 2026Article
- STARCall integrates image stitching, alignment, and read calling to enable scalable analysis of in situ sequencing data.PLoS computational biology · 2026Article
- Cell Type Populations for 3D Anatomical Structures of the Human Reference Atlas.Scientific data · 2026Article
- Single-molecule validation and optimised protocols for the use of secondary nanobodies in multiplexed immunoassays.Journal of microscopy · 2026Article
- Distinct spatial organization governs oral mucosal immunity.Nature immunology · 2026Article
38 more citing papers are in PubMed but not listed here.
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Authors and funding
17 authors at 4 institutions in 3 countries.
Funding
Abstract
High-content imaging is needed to catalog the variety of cellular phenotypes and multicellular ecosystems present in metazoan tissues. We recently developed iterative bleaching extends multiplexity (IBEX), an iterative immunolabeling and chemical bleaching method that enables multiplexed imaging (>65 parameters) in diverse tissues, including human organs relevant for international consortia efforts. IBEX is compatible with >250 commercially available antibodies and 16 unique fluorophores, and can be easily adopted to different imaging platforms using slides and nonproprietary imaging chambers. The overall protocol consists of iterative cycles of antibody labeling, imaging and chemical bleaching that can be completed at relatively low cost in 2-5 d by biologists with basic laboratory skills. To support widespread adoption, we provide extensive details on tissue processing, curated lists of validated antibodies and tissue-specific panels for multiplex imaging. Furthermore, instructions are included on how to automate the method using competitively priced instruments and reagents. Finally, we present a software solution for image alignment that can be executed by individuals without programming experience using open-source software and freeware. In summary, IBEX is a noncommercial method that can be readily implemented by academic laboratories and scaled to achieve high-content mapping of diverse tissues in support of a Human Reference Atlas or other such applications.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.