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The Auto-ICell system, a novel, and cost-effective integrated dropletmicrofluidic system, is introduced for real-time analysis of single-cellmorphology and apoptosis. This system integrates a 3D-printed microfluidic chipwith image analysis algorithms, enabling the generation of uniform dropletreactors and immediate image analysis. The system employs a color-based imageanalysis algorithm in the bright field for droplet content analysis. Meanwhile,in the fluorescence field, cell apoptosis is quantitatively measured through acombination of deep-learning-enabled multiple fluorescent channel analysis anda live/dead cell stain kit. Breast cancer cells are encapsulated within uniformdroplets, with diameters ranging from 70 {\mu}m to 240 {\mu}m, generated at ahigh throughput of 1,500 droplets per minute. Real-time image analysis resultsare displayed within 2 seconds on a custom graphical user interface (GUI). Thesystem provides an automatic calculation of the distribution and ratio ofencapsulated dyes in the bright field, and in the fluorescent field, cellblebbing and cell circularity are observed and quantified respectively. TheAuto-ICell system is non-invasive and provides online detection, offering arobust, time-efficient, user-friendly, and cost-effective solution forsingle-cell analysis. It significantly enhances the detection throughput ofdroplet single-cell analysis by reducing setup costs and improving operationalperformance. This study highlights the potential of the Auto-ICell system inadvancing biological research and personalized disease treatment, withpromising applications in cell culture, biochemical microreactors, drugcarriers, cell-based assays, synthetic biology, and point-of-care diagnostics.

Auto-ICell: An Accessible and Cost-Effective Integrative Droplet Microfluidic System for Real-Time Single-Cell Morphological and Apoptotic Analysis