dc.contributor.author | Lin, Jia-Ren | |
dc.contributor.author | Rashid, Rumana | |
dc.contributor.author | Maliga, Zoltan | |
dc.contributor.author | Izar, Benjamin | |
dc.contributor.author | Santagata, Sandro | |
dc.contributor.author | Du, Ziming | |
dc.contributor.author | Wang, Shu | |
dc.contributor.author | Aster, Jon | |
dc.contributor.author | Songer, Peter | |
dc.date.accessioned | 2019-12-18T16:57:29Z | |
dc.date.issued | 2019-09-18 | |
dc.identifier.citation | Du, Ziming, Jia-Ren Lin, Rumana Rashid, Zoltan Maliga, Shu Wang, Jon C. Aster, Benjamin Izar, Peter K. Sorger, and Sandro Santagata. 2019. Qualifying Antibodies for Image-based Immune Profiling and Multiplexed Tissue Imaging. Nature Protocols 14, no. 10: 2900-930. | en_US |
dc.identifier.issn | 1754-2189 | en_US |
dc.identifier.issn | 1750-2799 | en_US |
dc.identifier.uri | http://nrs.harvard.edu/urn-3:HUL.InstRepos:42068905 | * |
dc.description.abstract | Multiplexed tissue imaging enables precise, spatially resolved enumeration and characterization of cell types and states in human resection specimens. A growing number of methods applicable to formalin-fixed, paraffin-embedded (FFPE) tissue sections have been described, the majority of which rely on antibodies for antigen detection and mapping. This protocol provides step-by-step procedures for confirming the selectivity and specificity of antibodies used in fluorescence-based tissue imaging and for the construction and validation of antibody panels. Although the protocol is implemented using tissue-based cyclic immunofluorescence (t-CyCIF) as an imaging platform, these antibody-testing methods are broadly applicable. We demonstrate assembly of a 16-antibody panel for enumerating and localizing T cells and B cells, macrophages, and cells expressing immune checkpoint regulators. The protocol is accessible to individuals with experience in microscopy and immunofluorescence; some experience in computation is required for data analysis. A typical 30-antibody dataset for 20 FFPE slides can be generated within 2 weeks. | en_US |
dc.language.iso | en_US | en_US |
dc.publisher | Springer Science and Business Media LLC | en_US |
dc.relation | Nature Protocols | en_US |
dash.license | LAA | |
dc.subject | General Biochemistry, Genetics and Molecular Biology | en_US |
dc.title | Qualifying Antibodies for Image-Based Immune Profiling and Multiplexed Tissue Imaging | en_US |
dc.type | Journal Article | en_US |
dc.description.version | Accepted Manuscript | en_US |
dc.relation.journal | Nature Protocols | en_US |
dash.depositing.author | Aster, Jon | |
dash.waiver | 2019-05-23 | |
dc.date.available | 2019-12-18T16:57:29Z | |
dash.affiliation.other | Faculty of Arts & Sciences | en_US |
dash.funder.name | NCI Cancer Center Support | en_US |
dash.funder.name | National Institutes of Health | en_US |
dash.funder.name | Claudia Adams Barr Program for Innovative Cancer Research | en_US |
dash.funder.name | Molecular Biophysics Training | en_US |
dash.funder.award | P30-CA06516 | en_US |
dash.funder.award | U54-CA225088 | en_US |
dash.funder.award | U54-CA225088 | en_US |
dash.funder.award | U2C-CA233280 | en_US |
dash.funder.award | R41-CA224503 | en_US |
dash.funder.award | T32-GM008313 | en_US |
dash.funder.award | K08-CA222663 | en_US |
dc.identifier.doi | 10.1038/s41596-019-0206-y | |
dc.source.journal | Nat Protoc | |
dash.waiver.reason | Publishing with Nature Protocols and purchasing a creative commons license is not an available option. We will release the manuscript in pubmed central 6 months following publication. | en_US |
dash.source.volume | 14;10 | |
dash.source.page | 2900-2930 | |
dash.contributor.affiliated | Rashid, Rumana | |
dash.contributor.affiliated | Maliga, Zoltan | |
dash.contributor.affiliated | Santagata, Sandro | |
dash.contributor.affiliated | Aster, Jon | |
dash.contributor.affiliated | Lin, Jia-Ren | |
dash.contributor.affiliated | Izar, Benjamin | |