title: Imaging cytometry for counting circulating tumor cells: comparative analysis of the CellSearch vs ImageStream systems.
abstract: Circulating tumor cell (CTC) enumeration is important clinically for identifying prognostic and predictive factors in patients with solid cancers. The CellSearch device (Veridex) is an immunomagnetic CTC selection and enumeration system used in clinical practice. The ImageStream (Amnis) combines the strengths of flow cytometry and fluorescent microscopy in a single platform and has potential application for CTC counting. The performance in CTC enumeration was compared between the ImageStream and CellSearch systems. Various numbers of PANC-1 tumor cells were spiked into 7.5 mL of peripheral blood from a healthy donor. Before cell analysis by the ImageStream, tumor cell enrichment was performed by immunomagnetic selection with anti-EpPCAM. Anti-CD45 and anti-CK markers were used to discriminate between tumor cells and leukocytes. The ratios of tumor cells recovered from each dilution were calculated for both methods. The Wilcoxon rank test was applied to compare the results of the two methods and the reference value. The results of the two tested methods differed significantly from the reference value, but did not differ between them. Nevertheless, lower level of trueness and precision was observed in ImageStream when fewer numbers of CTCs were analyzed. Our results suggest that ImageStream platform for CTC enumeration has a potential value for the early diagnosis of disseminated disease, but needs an improvement of precision for the enumeration of low number of CTC.
authors.affiliation: Molecular Genetics Laboratory, Elche University Hospital, Elche, El Palmar, Spain.
authors.forename: Natividad
authors.initials: N
authors.lastname: López-Riquelme
authors.forename: Alfredo
authors.initials: A
authors.lastname: Minguela
authors.forename: Flori
authors.initials: F
authors.lastname: Villar-Permuy
authors.forename: Daniel
authors.initials: D
authors.lastname: Ciprian
authors.forename: Adela
authors.initials: A
authors.lastname: Castillejo
authors.forename: María-Rocio
authors.initials: MR
authors.lastname: Álvarez-López
authors.forename: José-Luis
authors.initials: JL
authors.lastname: Soto
mesh_terms: D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000276:immunology; D014408:Biomarkers, Tumor / Q000097:blood*; D015815:Cell Adhesion Molecules / Q000276:immunology; D002452:Cell Count / Q000379:methods; D045744:Cell Line, Tumor; D002469:Cell Separation; D000071858:Epithelial Cell Adhesion Molecule; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D017493:Leukocyte Common Antigens / Q000276:immunology; D007962:Leukocytes / Q000276:immunology; D009360:Neoplastic Cells, Circulating / Q000276:immunology*; D010190:Pancreatic Neoplasms / Q000097:blood*
chemical_list: D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D017493:Leukocyte Common Antigens; C516769:PTPRC protein, human
keywords: CellSearch; Circulating tumor cells; ImageStream; imaging cytometry