ChipCytometry for multiplexed detection of protein and mRNA markers on human FFPE tissue samples

Sebastian Jarosch; Jan Köhlen; Sabrina Wagner; Elvira D'Ippolito; Dirk H Busch

doi:10.1016/j.xpro.2022.101374

ChipCytometry for multiplexed detection of protein and mRNA markers on human FFPE tissue samples

STAR Protoc. 2022 May 11;3(2):101374. doi: 10.1016/j.xpro.2022.101374. eCollection 2022 Jun 17.

Authors

Sebastian Jarosch¹, Jan Köhlen¹, Sabrina Wagner¹, Elvira D'Ippolito¹, Dirk H Busch¹

Affiliation

¹ Institute for Medical Microbiology, Immunology and Hygiene, Technical University of Munich (TUM), 81675 Munich, Bavaria, Germany.

Abstract

In this protocol, we describe the use of ChipCytometry to combine RNA in situ hybridization and antibody staining for multiplexed tissue imaging of human formalin-fixed and paraffin-embedded tissue samples. The advantages of ChipCytometry are long-term storage for re-interrogation and advanced image quality by high dynamic range imaging of staining and background. A titrated pretreatment of tissue samples bypasses challenges because of the retrieval of antigens on coverslips and achieves an optimal staining quality at the minimal expense of tissue integrity. For complete details on the use and execution of this protocol, please refer to Jarosch et al. (2021).

Keywords: Antibody; In Situ Hybridization; Microscopy; Molecular Biology; Single Cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Formaldehyde*
Humans
In Situ Hybridization
RNA* / genetics
RNA, Messenger / genetics

Substances

RNA, Messenger
Formaldehyde
RNA