Multi-label versus multi-class classification of blood cells and their aggregates in microfluidic channels

Label-free single-cell imaging offers a scalable, non-invasive alternative to fluorescence-based cytometry, yet inferring molecular phenotypes directly from bright-field morphology remains challenging. We present a unified Deep Learning (DL) framework that jointly performs White Blood Cell (WBC) classification and continuous protein-expression regression from label-free Differential Phase Contrast (DPC) images. Our model employs a Hybrid architecture that fuses convolutional fine-grained texture features with transformer-based global representations through a learnable cross-branch gating module, enabling robust morpho-molecular inference from DPC images. To support downstream interpretability, we further incorporate a Large Language Model (LLM) that generates concise, biologically grounded summaries of the predicted cell states. Experiments on the Berkeley Single Cell Computational Microscopy (BSCCM) and Blood Cells Image benchmarks demonstrate strong performance, achieving a 91.3% WBC classification accuracy and a 0.72 Pearson correlation for CD16 expression regression on BSCCM. These results underscore the promise of label-free single-cell imaging for cost-effective hematological profiling, enabling simultaneous phenotype identification and quantitative biomarker estimation without fluorescent staining. The source code is available at https://github.com/saqibnaziir/Single-Cell-Phenotyping.

Bone marrow smear review remains important for acute myeloid leukemia (AML) assessment, but manual single-cell interpretation is labor-intensive and patient-level diagnosis requires aggregation of many cellular observations. We present a cell-to-patient deep learning pipeline for AML-assisted diagnosis from bone marrow smear images. The study included 258 patients from six anonymized centers, including a main cohort of 169 patients from Centers 1-3 and an external validation cohort of 89 patients from Centers 4-6. A 16-category cell annotation vocabulary was used to describe the global cellular composition, including granulocytic, monocytic, erythroid, lymphoid, eosinophilic, and other cells. Rather than identifying strict AML blasts or leukemic blasts, the model targets an expert-defined composite category termed Composite Blast-like Cells (CBLC), comprising N, N1, M, M1, R, R1, J, and J1 according to the project-wide morphological standard. A fixed YOLO-based segmentation module detected cells, predicted contours were matched to expert polygon annotations by contour IoU, and standardized single-cell crops were generated. An EfficientNet-B0 classifier was trained through a two-stage GT-to-YOLO and YOLO-to-YOLO strategy with class-imbalance correction, center-border regularization, and morphology-assisted supervision. Cell-level predictions were aggregated into patient-level CBLC ratios for AML-oriented diagnostic support. The pipeline achieved stable internal validation and maintained external generalization, with ensemble weighted F1-scores of 0.9076, 0.8696, and 0.9124 on Centers 4, 5, and 6, respectively.

Bone-marrow cytology is inherently structured: each cell belongs to a hematopoietic lineage, and many cell types lie on ordered maturation trajectories. Standard flat classifiers ignore this structure, treating a mild same-lineage confusion the same as a severe cross-lineage mistake and predicting only discrete labels. We propose HemaHier, an ordinal-hierarchical prediction head for a frozen or lightly adapted cytology foundation model. Its central component is a chain-conditioned maturity score that reads a single maturity value under a per-chain query, supervised only on biologically valid healthy chains, while dysplastic and off-chain cell types remain classes but are excluded from maturity supervision. Fine and lineage predictions are coupled through a shared posterior that guarantees hierarchical consistency, and a staged objective first stabilizes recognition, then adds lineage and maturity supervision. On three bone-marrow datasets under a shared ontology, HemaHier achieves competitive recognition while reducing biologically severe errors and adding a within-lineage maturity ordering that flat classifiers lack. Code is available at https://github.com/xmindflow/HemaHier.