Multiple Immunostainings with Different Epitope Retrievals—The FOLGAS Protocol

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 retri...

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Detalles Bibliográficos
Autores principales: von Schoenfeld, Anna, Bronsert, Peter, Poc, Michael, Fuller, Andrew, Filby, Andrew, Kraft, Stefan, Kurowski, Konrad, Sörensen, Kristin, Huber, Julia, Pfeiffer, Jens, Proietti, Michele, Stehl, Verena, Werner, Martin, Seidl, Maximilian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8745613/
https://www.ncbi.nlm.nih.gov/pubmed/35008649
http://dx.doi.org/10.3390/ijms23010223
Descripción
Sumario: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.

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