Multiple Immunostainings with Different Epitope Retrievals-The FOLGAS Protocol.

International journal of molecular sciences 2021 Dec 25

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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.

Citation

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. Multiple Immunostainings with Different Epitope Retrievals-The FOLGAS Protocol. International journal of molecular sciences. 2021 Dec 25;23(1)