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

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.