• Angelo, Michael
• Thompson, Steve
• West, Robert
• Fienberg, Harris
• Greenwald, Noah F.
• Angoshtari, Roshan
• Marquez, Diana
• Mccaffrey, Erin
• Vijayaragavan, Kausalia
• Galli, Stephen J.
• Montine, Thomas J.
• Risom, Tyler
• Nolan, Garry
• Kirkwood, David
• Bosse, Marc
• Kambham, Neeraja
• Keren, Leeat
• Wang, Jennifer
• Bendall, Sean C.

Abstract

Understanding tissue structure and function requires tools that quantify the expression of multiple proteins while preserving spatial information. Here, we describe MIBI-TOF (multiplexed ion beam imaging by time of flight), an instrument that uses bright ion sources and orthogonal time-of-flight mass spectrometry to image metal-tagged antibodies at subcellular resolution in clinical tissue sections. We demonstrate quantitative, full periodic table coverage across a five-log dynamic range, imaging 36 labeled antibodies simultaneously with histochemical stains and endogenous elements. We image fields of view up to 800 mum * 800 mum at resolutions down to 260 nm with sensitivities approaching single-molecule detection. We leverage these properties to interrogate intrapatient heterogeneity in tumor organization in triple-negative breast cancer, revealing regional variability in tumor cell phenotypes in contrast to a structured immune response. Given its versatility and sample back-compatibility, MIBI-TOF is positioned to leverage existing annotated, archival tissue cohorts to explore emerging questions in cancer, immunology, and neurobiology.

Topics

• impedance spectroscopy
• spectrometry
• time-of-flight mass spectrometry