Circulating tumor cells (CTCs), with their close association with cancer metastasis, the most aggressive feature of solid tumors, represent an important aspect of "liquid biopsy," which provides minimally- or noninvasive approaches for cancer detection and disease status monitoring. CTC analysis has shown the potential clinical applications in several cancer types and has been approved by FDA for clinical use in advanced breast, prostate, and colorectal cancer prognosis. In this chapter, we describe a CTC isolation method using a cell size and deformability-based system, Parsortix, and the immunofluorescence staining method to detect CTCs with both epithelial and mesenchymal features. We also describe a repeated fluorescence in situ hybridization (FISH) approach to detect alterations of multiple genomic regions on the same CTCs after immunofluorescence analysis. This approach allows the study of CTCs as a biomarker for cancer detection, prognosis, and therapeutic response monitoring, as well as the study of the heterogeneity of CTCs and cancers.
Keywords: Circulating tumor cell; Enumeration; Epithelial-to-mesenchymal transition; Fluorescence in situ hybridization; Immunofluorescence staining.