A novel solution for real-timein-situcell distribution monitoring in 3D bioprinting via fluorescence imaging.

3D bioprinting is rapidly evolving as a transformative technology for constructing biological tissues with precise cell and bioink placement. However, ensuring the quality and viability of bioprinted structures presents significant challenges, highlighting the need for advanced monitoring systems. Our study introduces a space-efficient, non-invasive approach for real-time,in-situmonitoring of cell dispersion in bioprinted constructs. Utilizing a novelin-situfluorescence microscopy technique, we employ nanoparticles for cell tagging and integrate a compact digital microscope into the bioprinter for layer-by-layer imaging, significantly saving space and weight to make the solution adaptable to any commercial bioprinter. This method enhancesin-situanalysis by combining data from the fluorescence system with conventional visible spectrum imaging. The synergy of these datasets provides a detailed method to examine cell dispersion and facilitates continuous monitoring during the bioprinting process. This allows for the immediate identification and correction of irregularities in cell deposition. Our approach aims to advance 3D bioprinting, setting new standards for the reliability and efficiency of bioprinted structures.