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