Integrated microfluidic platform for simultaneous blood typing and hemoglobin quantification at the point of care

• Integrated Multifunctional Microfluidic Disc : Developed a compact centrifugal microfluidic platform for simultaneous blood typing (ABO and Rh) and hemoglobin quantification at the point of care. • Automated Sample Handling : Utilized a cascading aliquoting mechanism and Sephadex G-10-treated siphon channels to ensure precise metering and reliable, power-free fluid control of whole blood samples. • Dual Diagnostic Capability : Combined gel column agglutination for blood typing and a colorimetric tetramethylbenzidine (TMB)-peroxidase assay for hemoglobin detection within a single self-contained device. • Image-Based Analysis : Integrated a low-cost camera with custom image-processing algorithms to enable fully automated, objective, and reproducible result interpretation for both tests. • Portable and Low-Cost Design : Achieved high diagnostic accuracy comparable to conventional methods while offering advantages such as minimal reagent use, reduced sample volume, and suitability for resource-limited settings. Accurate blood typing (BT) and hemoglobin (Hgb) quantification are critical for transfusion safety and anemia screening, yet existing approaches typically require centralized laboratory infrastructure and perform these assays independently. We report a centrifugal microfluidic disc that integrates ABO/Rh blood typing and quantitative hemoglobin analysis within a single disposable point-of-care platform. The system processes 12 µL of whole blood and completes the full analytical workflow in 130 s using a programmable rotational protocol. The platform incorporates a deterministic cascading aliquoting architecture that meters 2 µL per chamber. Whole-blood transfer is enabled by Sephadex–treated passive siphon valves achieving 100% priming success without external actuation. Blood typing is implemented via on-disc gel column agglutination, reproducing clinical separation behavior under centrifugal actuation. Hemoglobin quantification is performed using a TMB-based colorimetric assay covering the clinically relevant range of 5–15 g/dL. A low-cost imaging module coupled with a custom image-processing algorithm provides automated, operator-independent interpretation of both agglutination patterns and colorimetric intensity. Validation with 32 clinical samples demonstrated high diagnostic agreement with the reference tube-based method. At the individual reaction chamber level, automated agglutination detection achieved a sensitivity of 100% and a specificity of 97.7%. Hemoglobin measurements showed strong agreement with a clinical CBC analyzer (R² = 0.95), achieving 90.9% sensitivity and 80% specificity for anemia detection. By integrating deterministic metering, passive valving, gel-based immunoassay, biochemical quantification, and automated readout within a power-independent architecture, the proposed system establishes a compact and scalable framework for integrated hematologic point-of-care diagnostics.