Multiple Immunostainings with Different Epitope Retrievals—The FOLGAS Protocol
Authors:
- Anna von SchoenfeldASAnna von Schoenfeld
- Peter BronsertPBPeter Bronsert
- Michael PocMPMichael Poc
- Andrew FullerAFAndrew Fuller
- Andrew FilbyAFAndrew Filby
- Stefan KraftSKStefan Kraft
- Konrad KurowskiKKKonrad Kurowski
- Kristin SörensenKSKristin Sörensen
- Julia HuberJHJulia Huber
- Jens PfeifferJPJens Pfeiffer
- Michele ProiettiMPMichele Proietti
- Verena StehlVSVerena Stehl
Open Access
- 25 December 2021
- journal article
- research article
- Published by MDPI AG in International Journal of Molecular Sciences
- Vol. 23 (1) , 223
Abstract
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.Keywords
Funding Information
- Bundesministerium für Forschung, Technologie und Raumfahrt (BMBF 01 EO 0803)
- Deutsche Forschungsgemeinschaft (SFB1160 – IMPAT)
- Fördergesellschaft Forschung Tumorbiologie (Grant “Liquid-Biopsy-Initiative”)
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