Time-lapse or longitudinal fluorescence microscopy is broadly used in cell biology. However, current available time-lapse fluorescence microscopy systems are bulky and costly. The limited field-of-view (FOV) associated with microscope objective necessitates mechanical scanning if a larger FOV is required. Here we demonstrate a wide FOV time-lapse fluorescence self-imaging Petri dish system, termed the Talbot Fluorescence ePetri, which addresses these issues. This system’s imaging is accomplished through the use of the Fluorescence Talbot Microscopy (FTM). By incorporating a microfluidic perfusion subsystem onto the platform, we can image cell cultures directly from within an incubator. Our prototype has a resolution limit of 1.2 µm and an FOV of 13 mm2. As demonstration, we obtained time-lapse images of HeLa cells expressing H2B-eGFP. We also employed the system to analyze the cells’ dynamic response to an anticancer drug, camptothecin (CPT). This method can provide a compact and simple solution for automated fluorescence imaging of cell cultures in incubators.