A Do-It-Yourself and 3D-Printed Time-Lapse Imaging Platform for In-Incubator Live Cell Observation

Integrated in-incubator microscopy systems and all-in-one time-lapse culture incubators are often expensive and proprietary, which limits accessibility and customization. Open-source alternatives exist but often involve tradeoffs in imaging performance, illumination control, or adaptability. Here, we describe a do-it-yourself (DIY), low-cost, three-dimensional (3D)-printed time-lapse imaging platform designed for use inside standard cell culture incubators. The device employs a Raspberry Pi-based controller with stepper motor actuation and a programmable light-emitting diode (LED) backlight that enables both bright-field and oblique illumination modes. The system supports automated multi-well imaging, scheduled time-lapse acquisition, and real-time preview through open-source Python software. Performance validation demonstrated reliable long-term imaging of cultured cells and early embryos, with stable operation under incubator conditions. Optical resolution testing using a United States Air Force (USAF) 1951 target confirmed a minimum resolvable feature of approximately 1.55 µm. Motion evaluation showed low drift and high repeatability, with positioning accuracy closely matching commanded displacements. The system is compatible with multiple plate formats and can be adapted for specialized culture setups. Overall, this platform provides a cost-effective, reproducible, and customizable solution for long-term live-cell and embryo imaging in laboratory settings.