ArticleScientific reports2019
Ultra-fast and automated immunohistofluorescent multistaining using a microfluidic tissue processor.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers, 2 of them syntheses that pooled it.
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Who cites it
15 citing papers in PubMed, 2 syntheses or guidelines pooled it, 30 citations in OpenAlex.
- Society for Immunotherapy of Cancer: updates and best practices for multiplex immunohistochemistry (IHC) and immunofluorescence (IF) image analysis and data sharing.Journal for immunotherapy of cancer · 2025Guideline
- Research progress and perspectives on the application of tyramide signal amplification-based multiplex immunohistochemistry/immunofluorescence: a bibliometrics analysis.Frontiers in oncology · 2024Pooled it
- Spatial-Omics Methods and Applications.Methods in molecular biology (Clifton, N.J.) · 2025Review
- Increased Multiplexity in Optical Tissue Clearing-Based Three-Dimensional Immunofluorescence Microscopy of the Tumor Microenvironment by Light-Emitting Diode Photobleaching.Laboratory investigation; a journal of technical methods and pathology · 2024Article
- Semi-automated approaches for interrogating spatial heterogeneity of tissue samples.Scientific reports · 2024Article
- Fully automated sequential immunofluorescence (seqIF) for hyperplex spatial proteomics.Scientific reports · 2023Article
- High-plex protein and whole transcriptome co-mapping at cellular resolution with spatial CITE-seq.Nature biotechnology · 2023Article
- Spatial subcellular organelle networks in single cells.Scientific reports · 2023Article
- Spatial-CITE-seq: spatially resolved high-plex protein and whole transcriptome co-mapping.Research square · 2022Article
- Automated multimodal fluorescence microscopy for hyperplex spatial-proteomics: Coupling microfluidic-based immunofluorescence to high resolution, high sensitivity, three-dimensional analysis of histological slides.Frontiers in oncology · 2022Article
- SeqStain is an efficient method for multiplexed, spatialomic profiling of human and murine tissues.Cell reports methods · 2021Article
- A manual multiplex immunofluorescence method for investigating neurodegenerative diseases.Journal of neuroscience methods · 2020Article
- Spatially multiplexed RNA in situ hybridization to reveal tumor heterogeneity.Nucleic acids research · 2020Article
- Rapid micro-immunohistochemistry.Microsystems & nanoengineering · 2020Article
- Microfluidics-assisted multiplexed biomarker detection for in situ mapping of immune cells in tumor sections.Microsystems & nanoengineering · 2019Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Multistaining of a tissue section targeting multiple markers allows to reveal complex interplays in a tumor environment. However, the resource-intensive and impractically long nature of iterative multiplexed immunostainings prohibits its practical implementation in daily routine, even when using work-flow automation systems. Here, we report a fully automated and ultra-fast multistaining using a microfluidic tissue processor (MTP) in as short as 20 minutes per marker, by immunofluorescent staining employing commercially available tyramide signal amplification polymer precipitation by horse-radish peroxidase (HRP) activation. The reported duration includes (i) 15 minutes for the entire fluidic exchange and reagent incubation necessary for the immunostaining and (ii) 5 minutes for the heat-induced removal of the applied antibodies. Using the automated MTP, we demonstrated a 4-plex automated multistaining with clinically relevant biomarkers within 84 minutes, showing perfect agreement with the state-of-the-art microwave treatment antibody removal. The presented HRP-based method is in principle extendable to multistaining by both tyramides accommodating higher number of fluorescent channels and multi-color chromogenic staining. We anticipate that our automated multi-staining with a turn-around time shorter than existing monoplex immunohistochemistry methods has the potential to enable multistaining in routine without disturbing the current laboratory workflow, opening perspectives for implementation of -omics approaches in tissue diagnostics.
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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.