ArticleThe journal of histochemistry and cytochemistry : official journal of the Histochemistry Society2021
Improvement of Opal Multiplex Immunofluorescence Workflow for Human Tissue Sections.
Article in The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers, 1 of them a synthesis that pooled it.
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Who cites it
24 citing papers in PubMed, 1 synthesis or guideline pooled it, 27 citations in OpenAlex.
- Research progress and perspectives on the application of tyramide signal amplification-based multiplex immunohistochemistry/immunofluorescence: a bibliometrics analysis.Frontiers in oncology · 2024Pooled it
- Enhanced expression of ADAMTS1 in ovarian carcinomas: loss of ADAMTS1 expression instigates cellular reprogramming of extracellular matrix ensuing altered plasticity, augmented migration and attenuated adhesion.Journal of biomedical science · 2026Article
- Integrative spatial profiling pipeline for determining TME architectures in archival clinical specimens using CmTSA superplex technology.Cell discovery · 2026Article
- Mapping multipathology via spatial omic integration.Current opinion in biotechnology · 2026Review
- From Stains to Spectra: Multiplexed Immunohistochemistry and Mass Spectrometry Imaging for Spatial Proteomics in Biomedicine.Advances in experimental medicine and biology · 2026Review
- Article
- Density of T-cell Subsets in Colorectal Cancer in Relation to Disease-Specific Survival.Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology · 2025Article
- Optimization of a thermochemical antibody stripping method for enhanced tyramide signal amplification-based opal multiplex immunohistochemistry in fragile tissues.BMC research notes · 2025Article
- Exploring the characteristics of immortalized human ovarian surface epithelial cell lines.Heliyon · 2025Article
- Building Spatiotemporal Understanding ofACS infectious diseases · 2025Review
- Topological importance of CD8Annals of hematology · 2025Article
- Spatial Transcriptomics: Integrating Morphology and Molecular Mechanisms of Kidney Diseases.The American journal of pathology · 2025Review
- Nose-to-brain translocation of inhaled ultrafine elongated particles: facts and mysteries.Frontiers in toxicology · 2025Article
- Cell-mediated cytotoxicity within CSF and brain parenchyma in spinal muscular atrophy unaltered by nusinersen treatment.Nature communications · 2024Article
- Review
- Association between somatic microsatellite instability, hypermutation status, and specific T cell subsets in colorectal cancer tumors.Frontiers in immunology · 2024Article
- Multiplex Immunofluorescence: A Powerful Tool in Cancer Immunotherapy.International journal of molecular sciences · 2023Review
- ENPP1 deletion causes mouse osteoporosis via the MKK3/p38 MAPK/PCNA signaling pathway.Journal of orthopaedic surgery and research · 2022Article
- Cytotoxic CD8Scientific reports · 2022Article
- Navigating the cellular landscape in tissue: Recent advances in defining the pathogenesis of human disease.Computational and structural biotechnology journal · 2022Review
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The Opal multiplex technique is an established methodology for the detection of multiple biomarkers in one section. The protocol encompasses iterative single stainings and heating-mediated removal of the primary and secondary antibodies after each staining round, leaving untouched the Opal fluorophores which are deposited onto the antigen of interest. According to our experience, repetitive heating of skin sections often results in tissue damage, indicating an urgent need for milder alternatives to strip immunoglobulins. In this study, we demonstrate that considerable heating-related damage was found not only in skin but also in tissues of different origin, mostly characterized by low cell density. Importantly, the morphology remained fully intact when sections were repetitively exposed to β-mercaptoethanol-containing stripping buffer instead of multiple heating cycles. However, target epitopes appeared sensitive at a differential degree to multiple treatments with stripping buffer, as shown by loss in staining intensity, but in all cases, the staining intensity could be restored by increment of the primary antibody concentrations. Application of β-mercaptoethanol-containing stripping buffer instead of heating for antibody removal markedly improved the quality of the Opal multiplex technique, as a substantial higher number of differently colored cells could be visualized within a well-conserved morphological context.
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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.