Live label-retaining cancer cell (LRCC) undergoes asymmetric cell division with nonrandom chromosomal cosegregation (ACD-NRCC) in real time. (A): Materials and methods to detect LRCC undergoing ACD-NRCC (Materials and Methods). (B): Confocal microscopic cinematography. Following a single cell: at t = 0 minute, single cell labeled with Cy5 (green). t = 210 minutes, single cell, two nuclei (Blue-cyto9), and only one is retaining the Cy5. t = 600 minutes, two cells, two nuclei, and only one retains Cy5. Images were confirmed using confocal Z-stack images. (C): LRCC undergoing ACD-NRCC, showing a cell (t = 210) in a middle of mitosis, two nuclei within the same cytoplasmic space; however, only one nucleus retains the Cy5 DNA label. (Supporting Information
Fig. S2 and Video S1). Cyto9 is a vital dye; Cy5 is a fluorescent dye. Abbreviations: dUTP, 2′-deoxyuridine 5′-triphosphate; FACS, fluorescence-activated cell sorting.