Detection and Characterization of Circulating Tumor Cells Using Imaging Flow Cytometry—A Perspective Study

SIMPLE SUMMARY: Liquid biopsy is non-invasive approach used to prognose and monitor tumor progression based on the detection and examination of metastasis-related events found in the patients’ blood (such as circulating tumor cells (CTCs), extracellular vesicles, and circulating nucleic acids). Different ultrasensitive techniques are applied to study those events and the biology of tumor dissemination, which in the future might complement standard diagnostics. Here, we suggest that CTCs analysis could be improved by the usage of imaging flow cytometry, combining advantages of both standard flow cytometry (high-scale analysis) and microscopy (high resolution) to investigate detailed features of those cells. From this perspective, we discuss the potential of this technology in the CTC field and present representative images of CTCs from breast and prostate cancer patients analyzed with this method. ABSTRACT: Tumor dissemination is one of the most-investigated steps of tumor progression, which in recent decades led to the rapid development of liquid biopsy aiming to analyze circulating tumor cells (CTCs), extracellular vesicles (EVs), and circulating nucleic acids in order to precisely diagnose and monitor cancer patients. Flow cytometry was considered as a method to detect CTCs; however, due to the lack of verification of the investigated cells’ identity, this method failed to reach clinical utility. Meanwhile, imaging flow cytometry combining the sensitivity and high throughput of flow cytometry and image-based detailed analysis through a high-resolution microscope might open a new avenue in CTC technologies and provide an open-platform system alternative to CellSearch(®), which is still the only gold standard in this field. Hereby, we shortly review the studies on the usage of flow cytometry in CTC identification and present our own representative images of CTCs envisioned by imaging flow cytometry providing rationale that this novel technology might be a good tool for studying tumor dissemination, and, if combined with a high CTC yield enrichment method, could upgrade CTC-based diagnostics.