Multiple Immunostainings with Different Epitope Retrievals—The FOLGAS Protocol

oleh: Anna von Schoenfeld, Peter Bronsert, Michael Poc, Andrew Fuller, Andrew Filby, Stefan Kraft, Konrad Kurowski, Kristin Sörensen, Julia Huber, Jens Pfeiffer, Michele Proietti, Verena Stehl, Martin Werner, Maximilian Seidl

Format: Article
Diterbitkan: MDPI AG 2021-12-01

Deskripsi

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.