biostudies-literatureRoutt AHNational Heart, Lung, and Blood InstituteUniversity of Houston, United States3152https://www.ebi.ac.uk/biostudies/studies/S-EPMC9950070biostudies-literatureUnknown13(1)Changes in red blood cell (RBC) morphology distribution have emerged as a quantitative biomarker for the degradation of RBC functional properties during hypothermic storage. Previously published automated methods for classifying the morphology of stored RBCs often had insufficient accuracy and relied on proprietary code and datasets, making them difficult to use in many research and clinical applications. Here we describe the development and validation of a highly accurate open-source RBC morphology classification pipeline based on ensemble deep learning (DL). The DL-enabled pipeline utilized adaptive thresholding or semantic segmentation for RBC identification, a deep ensemble of four convolutional neural networks (CNNs) to classify RBC morphology, and Kalman filtering with Hungarian assiScientific reportsDeep ensemble learning enables highly accurate classification of stored red blood cell morphology.PMC9950070R01HL151858R01HL117329Lu MPiety NZYang NShevkoplyas SSRoutt AHfalseDeep ensemble learning enables highly accurate classification of stored red blood cell morphology.Changes in red blood cell (RBC) morphology distribution have emerged as a quantitative biomarker for the degradation of RBC functional properties during hypothermic storage. Previously published automated methods for classifying the morphology of stored RBCs often had insufficient accuracy and relied on proprietary code and datasets, making them difficult to use in many research and clinical applications. Here we describe the development and validation of a highly accurate open-source RBC morphology classification pipeline based on ensemble deep learning (DL). The DL-enabled pipeline utilized adaptive thresholding or semantic segmentation for RBC identification, a deep ensemble of four convolutional neural networks (CNNs) to classify RBC morphology, and Kalman filtering with Hungarian assi2023-01-01T00:00:00Z2023 Feb2025-04-26T20:43:39.16Z2025-04-06T16:31:18.377ZS-EPMC99500703682329810.1038/s41598-023-30214-w