ArticleThe journal of histochemistry and cytochemistry : official journal of the Histochemistry Society2017
Multiplex Staining by Sequential Immunostaining and Antibody Removal on Routine Tissue Sections.
Article in The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 98 papers.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
98 citing papers in PubMed, 173 citations in OpenAlex.
- Emerging technologies in the spatial proteomics landscape.Analytical and bioanalytical chemistry · 2026Review
- Histological Processing of Organoids for Immunostaining.Bio-protocol · 2026Article
- Integrated multi-omics identifies distinct macrophage alterations during progression of metabolic dysfunction-associated steatohepatitis.Nature genetics · 2026Article
- A quantitative pipeline for whole-mount deep imaging and analysis of multi-layered organoids across scales.eLife · 2026Article
- Optically switchable CLEAR probes enable rapid, biocompatible and high-efficiency fluorophore exchange for ultra-plex, high-resolution immunofluorescence imaging.Chemical science · 2026Article
- Protocol for iterative indirect immunofluorescence imaging of frozen mouse intestinal tissues.STAR protocols · 2026Article
- Article
- Multiplex Immunofluorescence Staining of Paraffin-Embedded Tissue: Coverslip Mounting of Tissue Sections Improves Z-axis Optical Resolution in Widefield Microscopy.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Unified platform for multiplex immunofluorescence across liver tissues and engineered models.eGastroenterology · 2026Article
- The normal human lymph node cell classification and landscape defined by high-dimensional spatial proteomics.PloS one · 2026Article
- Mapping safety in space: the emerging role of spatial transcriptomics in safe drug development.Frontiers in toxicology · 2026Review
- ST-FFPE-mIF: integrating spatial transcriptomics and multiplex immunofluorescence in formalin-fixed paraffin-embedded tissues using Stereo-seq.Genome biology · 2025Article
- Single-cell spatial proteomics.Histology and histopathology · 2025Review
- Optimization of a thermochemical antibody stripping method for enhanced tyramide signal amplification-based opal multiplex immunohistochemistry in fragile tissues.BMC research notes · 2025Article
- Harmonizing TUNEL with multiplexed iterative immunofluorescence enriches spatial contextualization of cell death.Cell reports methods · 2025Article
- Photobleaching alters the morphometric analysis of fluorescently labeled neurons and microglial cells.Pathology oncology research : POR · 2025Article
- Multimodal Mass Spectrometry Imaging in Atlas Building: A Review.Seminars in nephrology · 2024Review
- Ultraplex microscopy: versatile highly-multiplexed molecular labeling and imaging across scale and resolution.bioRxiv : the preprint server for biology · 2024Article
- A novel dual-release scaffold for fluorescent labels improves cyclic immunofluorescence.RSC chemical biology · 2024Article
- A spatial architecture-embedding HLA signature to predict clinical response to immunotherapy in renal cell carcinoma.Nature medicine · 2024Article
38 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
7 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Multiplexing, labeling for multiple immunostains in the very same cell or tissue section in situ, has raised considerable interest. The methods proposed include the use of labeled primary antibodies, spectral separation of fluorochromes, bleaching of the fluorophores or chromogens, blocking of previous antibody layers, all in various combinations. The major obstacles to the diffusion of this technique are high costs in custom antibodies and instruments, low throughput, and scarcity of specialized skills or facilities. We have validated a method based on common primary and secondary antibodies and diffusely available fluorescent image scanners. It entails rounds of four-color indirect immunofluorescence, image acquisition, and removal (stripping) of the antibodies, before another stain is applied. The images are digitally registered and the autofluorescence is subtracted. Removal of antibodies is accomplished by disulfide cleavage and a detergent or by a chaotropic salt treatment, this latter followed by antigen refolding. More than 30 different antibody stains can be applied to one single section from routinely fixed and embedded tissue. This method requires a modest investment in hardware and materials and uses freeware image analysis software. Multiplexing on routine tissue sections is a high throughput tool for in situ characterization of neoplastic, reactive, inflammatory, and normal cells.
Indexed as
Identifiers
What OpenQuestion holds
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.