Few-Cell Culture Monitoring and Analysis in Real-Time Using the Lensless Imaging System

For cell culture processes that are time-consuming to observe and analyze manually and prone to bacterial contamination. This paper presents a method for monitoring and realtime analysis of small numbers of cells in culture that combines lensless imaging techniques with microfluidics. The method uses microfluidics to create the microcavity environment required for cell growth, and images of the cells are taken at regular intervals using a lensless holographic imaging system. Information on the morphology, size, position, and number of cells is obtained by autofocus and holographic reconstruction of the collected hologram. A cell growth database is created based on all the image frame data. Finally, the growth database is post-processed to provide information such as cell growth and proliferation curves, trajectory tracking, and temperature and humidity profiles of the system environment. By monitoring the yeast in culture for 10 hours, a database of cell growth at 2-minute intervals was created, further growth curves for the yeast were obtained, and the cells were successfully tracked. The method's validity has been verified in this paper, providing a new solution for fully automated culture monitoring and analysis of small numbers of cells.