Figure 4: Automated scanning and tracking of single cells: (A) Lowresolution scan of the nano-volume well chip (fluorescently labeled with Rhodamine) to determine the localization of each well. (B) Identification of nano-volume wells containing single cells by detecting the presence or absence of single-spot fluorescence in each well. (C) High-resolution images for selected nano-volume wells containing a single cell. (D) Magnification of a high-resolution image showing one single cell in a well. (E) Microscopic images of four representative nano-volume wells taken on day 1 and day 7 after seeding of cells directly obtained from the digestion of a primary PDAC tumor. (F) Microscopic images of four nano-volume wells on day 1 and day 7 after seeding cells from a human blood sample spiked with GFP-positive primary PDAC tumor cells.