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Multiple Immunostainings with Different Epitope Retrievals-The FOLGAS Protocol.

ABSTRACT: We describe a sequential multistaining protocol for immunohistochemistry, immunofluorescence and CyTOF imaging for formalin-fixed, paraffin-embedded specimens (FFPE) in the formalin gas-phase (FOLGAS), enabling sequential multistaining, independent from the primary and secondary antibodies and retrieval. Histomorphologic details are preserved, and crossreactivity and loss of signal intensity are not detectable. Combined with a DAB-based hydrophobic masking of metal-labeled primary antibodies, FOLGAS allows the extended use of CyTOF imaging in FFPE sections.

SUBMITTER: von Schoenfeld A

PROVIDER: S-EPMC8745613 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature

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Multiple Immunostainings with Different Epitope Retrievals-The FOLGAS Protocol.

von Schoenfeld Anna A Bronsert Peter P Poc Michael M Fuller Andrew A Filby Andrew A Kraft Stefan S Kurowski Konrad K Sörensen Kristin K Huber Julia J Pfeiffer Jens J Proietti Michele M Stehl Verena V Werner Martin M Seidl Maximilian M

International journal of molecular sciences 20211225 1

We describe a sequential multistaining protocol for immunohistochemistry, immunofluorescence and CyTOF imaging for formalin-fixed, paraffin-embedded specimens (FFPE) in the formalin gas-phase (FOLGAS), enabling sequential multistaining, independent from the primary and secondary antibodies and retrieval. Histomorphologic details are preserved, and crossreactivity and loss of signal intensity are not detectable. Combined with a DAB-based hydrophobic masking of metal-labeled primary antibodies, FO ...[more]

PMID: 35008649

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