Multiplex protein detection on circulating tumor cells from liquid biopsies using imaging mass cytometry

Abstract

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.

Rare osteosarcoma cell subpopulation protein array and profiling using imaging mass cytometry and bioinformatics analysis

“…While our protocol solves the fast and multiplex analysis of rare cells, it is limited by the constraints of the first generation of IMC machines [ 18 , 23 ]. The primary limitation is the image resolution, which cannot exceed 1 μm per pixel and significant background noise and non-specific binding.…”

“…This can provide significantly greater protein quantification and co-localization information compared to conventional methods. However, the research on using IMC for rare cell study is barely seen except for one [ 23 ]. Our goal in this report is not only to utilize this IMC imaging method but also integrating bioinformatic analytical tools for rare cell characterization and profiling.…”

Diagnostic Potential of Imaging Flow Cytometry

“…Recently, multiplex protein detection on circulating tumor cells using imaging mass cytometry has been demonstrated [13].…”

Current Trends of Microfluidic Single-Cell Technologies

“…Erik Gerdtsson et al [ 225 ] showed that the integration of Imaging Mass Cytometry (IMC) using metal-labeled antibodies into the existing high definition single-cell analysis [ 226 ] allowed the CTC profiling to be extended to include lot more protein biomarkers. The IMC process involves the coupling of laser ablation with single-cell mass cytometry performed using the CyTOF technology [ 227 ].…”