Molecular analysis of circulating and disseminated tumor cells (CTCs/DTCs) has great potential as a means for continuous evaluation of prognosis and treatment efficacy in near-real time through minimally invasive liquid biopsies. To realize this potential, however, methods for molecular analysis of these rare cells must be developed and validated. Here, we describe the integration of imaging mass cytometry (IMC) using metal-labeled antibodies as implemented on the Fluidigm Hyperion Imaging System into the workflow of the previously established High Definition Single Cell Analysis (HD-SCA) assay for liquid biopsies, along with methods for image analysis and signal normalization. Using liquid biopsies from a metastatic prostate cancer case, we demonstrate that IMC can extend the reach of CTC characterization to include dozens of protein biomarkers, with the potential to understand a range of biological properties that could affect therapeutic response, metastasis and immune surveillance when coupled with simultaneous phenotyping of thousands of leukocytes.

Aerospace and Mechanical Engineering University of Southern California Los Angeles CA

Biological Sciences University of Southern California Los Angeles CA

Biomedical Center Faculty of Medicine in Pilsen Charles University Prague alej Svobody 76 323 00 Pilsen Czech Republic;

Biomedical Engineering University of Southern California Los Angeles CA

Department of Genitourinary Medical Oncology The University of Texas MD Anderson Cancer Center 1515 Holcombe Blvd Unit 207 Houston Texas;

Division of Hematology Department of Medicine USC Norris Comprehensive Cancer Center Keck School of Medicine University of Southern California 1450 Biggy Street Los Angeles CA;

Medicine University of Southern California Los Angeles CA

USC David and Dana Dornsife College of Letters Arts and Sciences 3430 S Vermont Ave TRF 114 MC3303 Los Angeles CA 90089 3303;

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