Detecting glucose levels is crucial for diabetes patients as it enables timely and effective management, preventing complications and promoting overall health. In this endeavor, we have designed a novel, affordable point-of-care diagnostic device utilizing microfluidic principles, a smartphone camera, and established laboratory colorimetric methods for accurate glucose estimation. Our proposed microfluidic device comprises layers of adhesive poly-vinyl films stacked on a poly methyl methacrylate (PMMA) base sheet, with micro-channel contours precision-cut using a cutting printer. Employing the gold standard glucose-oxidase/peroxidase reaction on this microfluidic platform, we achieve enzymatic glucose determination. The resulting colored complex, formed by phenol and 4-aminoantipyrine in the presence of hydrogen peroxide generated during glucose oxidation, is captured at various glucose concentrations using a smartphone camera. Raw images are processed and utilized as input data for a 2-D convolutional neural network (CNN) deep learning classifier, demonstrating an impressive 95% overall accuracy against new images. The glucose predictions done by CNN are compared with ISO 15197:2013/2015 gold standard norms. Furthermore, the classifier exhibits outstanding precision, recall, and F1 score of 94%, 93%, and 93%, respectively, as validated through our study, showcasing its exceptional predictive capability. Next, a user-friendly smartphone application named “GLUCOLENS AI” was developed to capture images, perform image processing, and communicate with cloud server containing the CNN classifier. The developed CNN model can be successfully used as a pre-trained model for future glucose concentration predictions.© 2024. The Author(s).