Microfluidic Light-Scattering Imaging Coupled with Deep Learning for Label-Free Single-Cell Classification of...

Accurate classification of lymphoma cell subtypes is essential for disease diagnosis and therapeutic decision-making, yet conventional approaches often rely on fluorescence labeling, labor-intensive sample preparation, and specialized instrumentation, limiting their applicability for rapid, label-free single-cell analysis. Here, we present an AI-assisted microfluidic light-scattering imaging platform for label-free classification of lymphoma cells. The platform integrates hydrodynamic focusing within a microfluidic chip, continuous acquisition of two-dimensional (2D) light-scattering patterns, automated image preprocessing, and transfer learning based on a pretrained ResNet50 network for intelligent optical feature extraction and classification. Human B lymphoma (Daudi) and T lymphoblastic lymphoma (SUP-T1) cells were used to evaluate the proposed framework. The optical imaging system was first validated using standard microspheres, demonstrating reliable acquisition of light-scattering patterns under continuous-flow conditions. A dataset comprising 800 single-cell scattering patterns was subsequently established and evaluated using stratified five-fold cross-validation. The proposed framework achieved an average classification accuracy of 94.75% with an average area under the receiver operating characteristic (ROC) curve of 0.986. By integrating microfluidic optical biosensing with deep learning, this work enables automated interpretation of intrinsic optical scattering signatures and provides a promising AI-enabled strategy for rapid, label-free lymphoma screening and intelligent healthcare applications.