Multiple Immunostainings with Different Epitope Retrievals-The FOLGAS Protocol.
Anna von SchoenfeldPeter BronsertMichael PocAndrew FullerAndrew FilbyStefan KraftKonrad KurowskiKristin SörensenJulia HuberJens PfeifferMichele ProiettiVerena StehlMartin WernerMaximilian SeidlPublished in: International journal of molecular sciences (2021)
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.