International Journal of Molecular Sciences (Dec 2021)

Multiple Immunostainings with Different Epitope Retrievals—The FOLGAS Protocol

  • 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

Journal volume & issue
Vol. 23, no. 223
p. 223


Read online

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.